WO2021206636A1 - ANTIGEN-BINDING PROTEIN TO SARS-CoV-2 - Google Patents
ANTIGEN-BINDING PROTEIN TO SARS-CoV-2 Download PDFInfo
- Publication number
- WO2021206636A1 WO2021206636A1 PCT/SG2021/050195 SG2021050195W WO2021206636A1 WO 2021206636 A1 WO2021206636 A1 WO 2021206636A1 SG 2021050195 W SG2021050195 W SG 2021050195W WO 2021206636 A1 WO2021206636 A1 WO 2021206636A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- chain
- heavy
- light
- antigen
- Prior art date
Links
- 241001678559 COVID-19 virus Species 0.000 title claims abstract description 175
- 108090000623 proteins and genes Proteins 0.000 title description 6
- 102000004169 proteins and genes Human genes 0.000 title description 5
- 102000025171 antigen binding proteins Human genes 0.000 claims abstract description 202
- 108091000831 antigen binding proteins Proteins 0.000 claims abstract description 202
- 229940096437 Protein S Drugs 0.000 claims abstract description 46
- 238000000034 method Methods 0.000 claims abstract description 45
- 101710198474 Spike protein Proteins 0.000 claims abstract description 44
- 230000003472 neutralizing effect Effects 0.000 claims abstract description 28
- 102000005962 receptors Human genes 0.000 claims abstract description 14
- 108020003175 receptors Proteins 0.000 claims abstract description 14
- 102100035360 Cerebellar degeneration-related antigen 1 Human genes 0.000 claims description 193
- 230000035772 mutation Effects 0.000 claims description 45
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims description 39
- 241000315672 SARS coronavirus Species 0.000 claims description 34
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 claims description 22
- 239000002773 nucleotide Substances 0.000 claims description 15
- 125000003729 nucleotide group Chemical group 0.000 claims description 15
- 239000008194 pharmaceutical composition Substances 0.000 claims description 11
- 241000494545 Cordyline virus 2 Species 0.000 claims description 9
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims description 8
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 8
- 230000010807 negative regulation of binding Effects 0.000 claims description 8
- 230000007910 cell fusion Effects 0.000 claims description 7
- 230000004927 fusion Effects 0.000 claims description 7
- 238000002560 therapeutic procedure Methods 0.000 claims description 7
- 230000005764 inhibitory process Effects 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 5
- 230000001404 mediated effect Effects 0.000 claims description 5
- 239000000032 diagnostic agent Substances 0.000 claims description 4
- 229940039227 diagnostic agent Drugs 0.000 claims description 4
- 239000003937 drug carrier Substances 0.000 claims description 4
- 241001515965 unidentified phage Species 0.000 claims description 3
- 208000025721 COVID-19 Diseases 0.000 abstract description 12
- 208000037847 SARS-CoV-2-infection Diseases 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 description 67
- 101000629318 Severe acute respiratory syndrome coronavirus 2 Spike glycoprotein Proteins 0.000 description 28
- 241000700605 Viruses Species 0.000 description 26
- 102000036639 antigens Human genes 0.000 description 22
- 108091007433 antigens Proteins 0.000 description 22
- 239000003795 chemical substances by application Substances 0.000 description 22
- 239000002245 particle Substances 0.000 description 22
- 239000000427 antigen Substances 0.000 description 20
- 230000000903 blocking effect Effects 0.000 description 20
- 150000001875 compounds Chemical class 0.000 description 20
- 238000003556 assay Methods 0.000 description 18
- 239000000523 sample Substances 0.000 description 18
- 125000003275 alpha amino acid group Chemical group 0.000 description 16
- 238000012575 bio-layer interferometry Methods 0.000 description 16
- 238000002965 ELISA Methods 0.000 description 14
- 101100454807 Caenorhabditis elegans lgg-1 gene Proteins 0.000 description 13
- 238000006386 neutralization reaction Methods 0.000 description 13
- 238000002823 phage display Methods 0.000 description 13
- 239000012634 fragment Substances 0.000 description 12
- 239000000203 mixture Substances 0.000 description 12
- 241000711573 Coronaviridae Species 0.000 description 10
- 239000012530 fluid Substances 0.000 description 10
- 241001112090 Pseudovirus Species 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- 230000003612 virological effect Effects 0.000 description 9
- 108090000288 Glycoproteins Proteins 0.000 description 8
- 102000003886 Glycoproteins Human genes 0.000 description 8
- 108091005634 SARS-CoV-2 receptor-binding domains Proteins 0.000 description 8
- 231100000673 dose–response relationship Toxicity 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 108091033319 polynucleotide Proteins 0.000 description 7
- 239000002157 polynucleotide Substances 0.000 description 7
- 102000040430 polynucleotide Human genes 0.000 description 7
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 description 6
- 102000053723 Angiotensin-converting enzyme 2 Human genes 0.000 description 6
- 108090000975 Angiotensin-converting enzyme 2 Proteins 0.000 description 6
- 238000013357 binding ELISA Methods 0.000 description 6
- 230000004071 biological effect Effects 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 208000015181 infectious disease Diseases 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 230000003389 potentiating effect Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 230000001225 therapeutic effect Effects 0.000 description 5
- 101710091045 Envelope protein Proteins 0.000 description 4
- 241000124008 Mammalia Species 0.000 description 4
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 description 4
- 101710188315 Protein X Proteins 0.000 description 4
- 102100021696 Syncytin-1 Human genes 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 230000002452 interceptive effect Effects 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000000241 respiratory effect Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 108010083359 Antigen Receptors Proteins 0.000 description 3
- 102000006306 Antigen Receptors Human genes 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 101000929928 Homo sapiens Angiotensin-converting enzyme 2 Proteins 0.000 description 3
- 244000309467 Human Coronavirus Species 0.000 description 3
- 241001529936 Murinae Species 0.000 description 3
- 102000004142 Trypsin Human genes 0.000 description 3
- 108090000631 Trypsin Proteins 0.000 description 3
- 230000010530 Virus Neutralization Effects 0.000 description 3
- 239000012472 biological sample Substances 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 102000048657 human ACE2 Human genes 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 238000003670 luciferase enzyme activity assay Methods 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- 239000012588 trypsin Substances 0.000 description 3
- 239000003981 vehicle Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 2
- 102100031673 Corneodesmosin Human genes 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 2
- 238000012286 ELISA Assay Methods 0.000 description 2
- 101710114810 Glycoprotein Proteins 0.000 description 2
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 2
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 2
- 108060001084 Luciferase Proteins 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 2
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 2
- 206010036790 Productive cough Diseases 0.000 description 2
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 2
- 101710167605 Spike glycoprotein Proteins 0.000 description 2
- 108010031318 Vitronectin Proteins 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000034217 membrane fusion Effects 0.000 description 2
- 108010087904 neutravidin Proteins 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000000159 protein binding assay Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 210000003802 sputum Anatomy 0.000 description 2
- 208000024794 sputum Diseases 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 231100000607 toxicokinetics Toxicity 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- YRNWIFYIFSBPAU-UHFFFAOYSA-N 4-[4-(dimethylamino)phenyl]-n,n-dimethylaniline Chemical compound C1=CC(N(C)C)=CC=C1C1=CC=C(N(C)C)C=C1 YRNWIFYIFSBPAU-UHFFFAOYSA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 208000010470 Ageusia Diseases 0.000 description 1
- 206010002653 Anosmia Diseases 0.000 description 1
- 101100454808 Caenorhabditis elegans lgg-2 gene Proteins 0.000 description 1
- 101100217502 Caenorhabditis elegans lgg-3 gene Proteins 0.000 description 1
- 241000282465 Canis Species 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010008177 Fd immunoglobulins Proteins 0.000 description 1
- 102000004961 Furin Human genes 0.000 description 1
- 108090001126 Furin Proteins 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 241000714192 Human spumaretrovirus Species 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 208000025370 Middle East respiratory syndrome Diseases 0.000 description 1
- 241000699729 Muridae Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 208000000112 Myalgia Diseases 0.000 description 1
- 108091007491 NSP3 Papain-like protease domains Proteins 0.000 description 1
- 206010028817 Nausea and vomiting symptoms Diseases 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102000016979 Other receptors Human genes 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 208000036071 Rhinorrhea Diseases 0.000 description 1
- 206010039101 Rhinorrhoea Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000710961 Semliki Forest virus Species 0.000 description 1
- 241000701093 Suid alphaherpesvirus 1 Species 0.000 description 1
- 101710120037 Toxin CcdB Proteins 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 235000019666 ageusia Nutrition 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000008512 biological response Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 239000000816 peptidomimetic Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000001138 tear Anatomy 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 210000001944 turbinate Anatomy 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
- C07K16/1002—Coronaviridae
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
- C07K16/1002—Coronaviridae
- C07K16/1003—Severe acute respiratory syndrome coronavirus 2 [SARS‐CoV‐2 or Covid-19]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/55—Fab or Fab'
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
Definitions
- the present disclosure relates broadly to antigen-binding protein capable of binding to SARS-CoV-2, and related products and methods.
- SARS-CoV-2 a novel coronavirus, designated as SARS-CoV-2, emerged in Wuhan, China.
- SARS-CoV-2 severe acute respiratory syndrome coronavirus [SARS-CoV] and Middle East respiratory syndrome coronavirus [MERS-CoV]
- SARS-CoV-2 also known as COVID-19, causes acute respiratory disease that is often severe.
- SARS-CoV-2 As of March 2021 , nearly 120 million individuals worldwide have been confirmed to have COVID-19 infection with more than 2 million fatalities occurred.
- SARS-CoV-2 is the third coronavirus to emerge in the human population in the past two decades — an emergence that has put global public health institutions on high alert.
- detecting agents and therapeutic agents may enhance detection, and also provide immediate protection and effective treatment which are urgently needed.
- an agent such as an antigen-binding protein, that is capable of binding and/or neutralising SARS-CoV- 2, and related products and methods.
- an isolated antigen-binding protein that is capable of binding to and/or neutralizing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
- the antigen-binding protein is capable of binding to a spike protein of SARS-CoV-2, optionally a receptor-binding domain (RBD) of SARS-CoV-2.
- the antigen-binding protein comprises one or more of the following properties:
- the antigen-binding protein does not cross-react with severe acute respiratory syndrome coronavirus (SARS-CoV). In one embodiment, the antigen-binding protein is capable of binding to SARS-CoV.
- SARS-CoV severe acute respiratory syndrome coronavirus
- the antigen-binding protein comprises a monoclonal antigen-binding protein.
- the antigen-binding protein comprises a fully human antigen-binding protein.
- the antigen-binding protein comprises an IgG-like structure.
- the antigen-binding protein comprises a Fab fragment.
- the antigen-binding protein is capable of binding to and/or neutralizing about two or more of the following SARS-CoV-2: a SARS- CoV-2 comprising a wild-type spike protein, a SARS-CoV-2 comprising a mutated spike protein with no more than about two mutations, a SARS-CoV-2 comprising a mutated spike protein with no more than about one mutation, a SARS-CoV-2 comprising a N439K mutation, a SARS-CoV-2 comprising a V483A mutation, a SARS-CoV-2 comprising a G476S mutation, a SARS-CoV-2 comprising a S494P mutation, a SARS-CoV-2 comprising a V483I mutation, a SARS-CoV-2 comprising L455I and F456V mutations and a SARS-CoV-2 comprising a D614G mutation.
- SARS-CoV-2 comprising a wild-type spike protein
- the antigen-binding protein is capable of inhibiting virus-cell fusion and/or cell-cell fusion mediated by SARS-CoV-2.
- the antigen-binding protein comprises: a light-chain complementarity determining region CDR1 selected from the group consisting of: SEQ ID NO: 28-50 and conservative sequence variants thereof; a light-chain CDR2 selected from the group consisting of: SEQ ID NO: 51 - 64 and conservative sequence variants thereof; a light-chain CDR3 selected from the group consisting of: SEQ ID NO: 65- 90 and conservative sequence variants thereof; a heavy-chain CDR 1 selected from the group consisting of: SEQ ID NO: 117-132 and conservative sequence variants thereof; a heavy-chain CDR 2 selected from the group consisting of: SEQ ID NO: 133-148 and conservative sequence variants thereof; and a heavy-chain CDR 3 selected from the group consisting of: SEQ ID NO: 149-173 and conservative sequence variants thereof.
- a light-chain complementarity determining region CDR1 selected from the group consisting of: SEQ ID NO: 28-50 and conservative sequence variants thereof
- a light-chain CDR2 selected from
- the antigen-binding protein comprises the following CDR sequences:
- light-chain CDR1 SGSIASHY (SEQ ID NO: 28)
- light-chain CDR2 EDN (SEQ ID NO: 51)
- light-chain CDR3 QSYGSGFVV (SEQ ID NO: 65)
- heavy-chain CDR1 GFTFSSYA (SEQ ID NO: 117)
- heavy-chain CDR2 ISGSGGST (SEQ ID NO: 133)
- heavy-chain CDR3 AKDYFRWL (SEQ ID NO: 149);
- C2 light-chain CDR1 QSLLHSNGFTY (SEQ ID NO: 32) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQGIQTPLT (SEQ ID NO: 69) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSSAGTFYFDY (SEQ ID NO: 153);
- light-chain CDR1 SGSIASYY (SEQ ID NO: 33) light-chain CDR2: EDD (SEQ ID NO: 54) light-chain CDR3: QSFDSSIQHVV (SEQ ID NO: 70) heavy-chain CDR1 : GYPFSTYY (SEQ ID NO: 121) heavy-chain CDR2: IDPSGGST (SEQ ID NO: 137) heavy-chain CDR3: ARVAEQHLDYYFDY (SEQ ID NO: 154);
- F4 light-chain CDR1 SSNIGRNF (SEQ ID NO: 36) light-chain CDR2: RNN (SEQ ID NO: 56) light-chain CDR3: QSYDNSLFVV (SEQ ID NO: 73) heavy-chain CDR1 : GDSVSSNSAA (SEQ ID NO: 122) heavy-chain CDR2: TYYRSKWYN (SEQ ID NO: 139) heavy-chain CDR3: AREKGIEGPAFDP (SEQ ID NO: 157);
- light-chain CDR1 SGSIASNY (SEQ ID NO: 34)
- light-chain CDR2 EDN (SEQ ID NO: 51)
- light-chain CDR3 QSYDENIRV (SEQ ID NO: 74)
- heavy-chain CDR1 GGSFSGYY (SEQ ID NO: 123)
- heavy-chain CDR2 INHSGST (SEQ ID NO: 140)
- heavy-chain CDR3 ARGRWLRGAFDI (SEQ ID NO: 158);
- 2C9 light-chain CDR1 QTVGGSY (SEQ ID NO: 37) light-chain CDR2: GAS (SEQ ID NO: 57) light-chain CDR3: QQYASSRT (SEQ ID NO: 75) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPILGIA (SEQ ID NO: 141) heavy-chain CDR3: ARHGGVGATTGYYYMDV (SEQ ID NO: 159); 2C10 light-chain CDR1 : SSTIGTNY (SEQ ID NO: 38) light-chain CDR2: DNY (SEQ ID NO: 58) light-chain CDR3: GTWDSRLSVGV (SEQ ID NO: 76) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INYRGNT (SEQ ID NO: 142) heavy-chain CDR3: ARWDGGNSVDH (SEQ ID NO: 160);
- 2C12 light-chain CDR1 QSINIY (SEQ ID NO: 39) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYITPQT (SEQ ID NO: 77) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARGGRFHYYYYAMDV (SEQ ID NO: 161);
- 2D3 light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ASSGWLRGAFDI (SEQ ID NO: 162);
- 2G7 light-chain CDR1 GGTIGSNY (SEQ ID NO: 40) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNRV (SEQ ID NO: 79) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGKWLRGAFDI (SEQ ID NO: 163); 2H4 light-chain CDR1 : EFIGDD (SEQ ID NO: 41) light-chain CDR2: EAR (SEQ ID NO: 60) light-chain CDR3: LQYDSFPLT (SEQ ID NO: 80) heavy-chain CDR1 : GYTFSGYY (SEQ ID NO: 124) heavy-chain CDR2: VNPNSGGT (SEQ ID NO: 144) heavy-chain CDR3: ARGGRPGLPAAGYIDY (SEQ ID NO: 164
- light-chain CDR1 SSDVGTYNY (SEQ ID NO: 42) light-chain CDR2: DVS (SEQ ID NO: 61) light-chain CDR3: ASFSSSSTLLV (SEQ ID NO: 81) heavy-chain CDR1 : GYTFSTYD (SEQ ID NO: 125) heavy-chain CDR2: ISTYNGDT (SEQ ID NO: 145) heavy-chain CDR3: ARGDSSVGYEYFQH (SEQ ID NO: 165);
- 3C5 light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGSLSGYY (SEQ ID NO: 126) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
- 3D2 light-chain CDR1 SGSIARNY (SEQ ID NO: 43) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDRTNKRV (SEQ ID NO: 82) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166); 3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO:
- light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNLQWV (SEQ ID NO: 84) heavy-chain CDR1 : GYTFTSYG (SEQ ID NO: 127) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARGIITMISDY (SEQ ID NO: 167);
- 3F1 light-chain CDR1 GGSIADNF (SEQ ID NO: 45) light-chain CDR2: DYS (SEQ ID NO: 62) light-chain CDR3: QSYDISNPV (SEQ ID NO: 85) heavy-chain CDR1 : GFTFSSYG (SEQ ID NO: 128) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARDGGGGMDV (SEQ ID NO: 168);
- 3F11 light-chain CDR1 HSDGRNY (SEQ ID NO: 46) light-chain CDR2: DTS (SEQ ID NO: 63) light-chain CDR3: HQFRRSVST (SEQ ID NO: 86) heavy-chain CDR1 : GYTFSSSG (SEQ ID NO: 129) heavy-chain CDR2: ISTYNGNT (SEQ ID NO: 146) heavy-chain CDR3: ATSIAVAGIDY (SEQ ID NO: 169); 3H7 light-chain CDR1 : TDSIASNY (SEQ ID NO: 47) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDGVNRGVI (SEQ ID NO: 87) heavy-chain CDR1 : GFTFSNYG (SEQ ID NO: 130) heavy-chain CDR2: IWHDGTNK (SEQ ID NO: 147) heavy-chain CDR3: VRDQGAGVWNGYSY (SEQ ID NO: 1
- light-chain CDR1 SGNIASYY (SEQ ID NO: 48)
- light-chain CDR2 EDN (SEQ ID NO: 51)
- light-chain CDR3 QSYDRPNHVV (SEQ ID NO: 88)
- heavy-chain CDR1 GYTFTSYY (SEQ ID NO: 119)
- heavy-chain CDR2 INPSGGST (SEQ ID NO: 135)
- heavy-chain CDR3 ASSVVPAAIYDYYYGMDV (SEQ ID NO: 171);
- light-chain CDR1 QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172);
- light-chain CDR1 QTINNNY (SEQ ID NO: 50) light-chain CDR2: DAS (SEQ ID NO: 64) light-chain CDR3: QQYGSSPRT (SEQ ID NO: 90) heavy-chain CDR1 : GFTVSSNY (SEQ ID NO: 132) heavy-chain CDR2: IYSGGST (SEQ ID NO: 148) heavy-chain CDR3: ARDYGDYYFDY (SEQ ID NO: 173); or conservative sequence variants thereof.
- the antigen-binding protein comprises a light-chain variable region comprising any one of SEQ ID NO: 1 -27 or a sequence sharing at least about 75% sequence similarity thereto; and/or a heavy-chain variable region comprising any one of SEQ ID NO: 91 -116 or a sequence sharing at least about 75% sequence similarity thereto.
- a first antigen-binding protein comprises the following CDR sequences:
- 5A6 light-chain CDR1 QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172) and a second antigen-binding protein comprises the following CDR sequences:
- 3D11 light-chain CDR1 SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
- a pharmaceutical composition comprising embodiments of the antigen-binding protein or embodiments of the combination, and a pharmaceutically acceptable carrier.
- embodiments of the antigen-binding protein embodiments of the combination or embodiments of the pharmaceutical composition, for use in therapy, or as a diagnostic agent.
- nucleotide sequence encoding for embodiments of the antigen-binding protein or embodiments of the combination.
- the nucleotide sequence comprises one of more of SEQ ID NO: 174-351 or a sequence sharing at least about 75% sequence identity thereto.
- a bacteriophage a host cell or a non-human organism comprising embodiments of the nucleotide sequence.
- a method of treating SARS-CoV-2 in a subject comprising administering to the subject embodiments of the antigen-binding protein, embodiments of the combination or embodiments of the pharmaceutical composition.
- the method comprises administering to the subject embodiments of the combination.
- a method of detecting the presence of SARS-CoV-2 in a sample comprising: contacting the sample with the embodiments of the antigen-binding protein; detecting a binding event involving the antigen-binding protein; wherein the detection of a binding event is indicative of the presence of SARS-CoV-2 in the sample.
- treating refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) a medical condition, which includes but is not limited to diseases (such as SARS-CoV-2 infections), symptoms and disorders.
- a medical condition also includes a body’s response to a disease or disorder, e.g., inflammation.
- Those in need of such treatment include those already with a medical condition as well as those prone to getting the medical condition or those in whom a medical condition is to be prevented.
- the term "therapeutically effective amount” will be an amount of an active agent that is capable of preventing, reversing or at least slowing down (lessening) a medical condition, such as SARS-CoV-2 infections
- Dosages and administration of agents, compounds, compositions and formulations of the present disclosure may be determined by one of ordinary skill in the art of clinical pharmacology or pharmacokinetics. See, for example, Mordenti and Rescigno, (1992) Pharmaceutical Research. 9:17-25; Morenti et al., (1991) Pharmaceutical Research. 8:1351 -1359; and Mordenti and Chappell, "The use of interspecies scaling in toxicokinetics" in Toxicokinetics and New Drug Development, Yacobi et al.
- an effective amount of the active agent of the present disclosure to be employed therapeutically will depend, for example, upon the therapeutic objectives, the route of administration, and the condition of the subject. Accordingly, it may be necessary for the therapist to titer the dosage and modify the route of administration as required to obtain the optimal therapeutic effect.
- subject as used herein includes patients and non-patients.
- patient refers to individuals suffering or are likely to suffer from a medical condition such as a SARS-CoV-2 infection
- non-patients refer to individuals not suffering and are likely to not suffer from the medical condition.
- Non-patients include healthy individuals, non-diseased individuals and/or an individual free from the medical condition.
- subject includes humans and animals. Animals may include, but is not limited to, mammals (for example non-human primates, canine, murine and the like), and the like.
- “Murine” refers to any mammal from the family Muridae, such as mouse, rat, rabbit, and the like.
- micro as used herein is to be interpreted broadly to include dimensions from about 1 micron to about 1000 microns.
- nano as used herein is to be interpreted broadly to include dimensions less than about 1000 nm.
- the term “particle” as used herein broadly refers to a discrete entity or a discrete body.
- the particle described herein can include an organic, an inorganic or a biological particle.
- the particle used described herein may also be a macro particle that is formed by an aggregate of a plurality of sub-particles or a fragment of a small object.
- the particle of the present disclosure may be spherical, substantially spherical, or non-spherical, such as irregularly shaped particles or ellipsoidally shaped particles.
- size when used to refer to the particle broadly refers to the largest dimension of the particle. For example, when the particle is substantially spherical, the term “size” can refer to the diameter of the particle; or when the particle is substantially non-spherical, the term “size” can refer to the largest length of the particle.
- Coupled or “connected” as used in this description are intended to cover both directly connected or connected through one or more intermediate means, unless otherwise stated.
- association with refers to a broad relationship between the two elements.
- the relationship includes, but is not limited to a physical, a chemical or a biological relationship.
- elements A and B may be directly or indirectly attached to each other or element A may contain element B or vice versa.
- adjacent refers to one element being in close proximity to another element and may be but is not limited to the elements contacting each other or may further include the elements being separated by one or more further elements disposed therebetween.
- the word “substantially” whenever used is understood to include, but not restricted to, “entirely” or “completely” and the like.
- terms such as “comprising”, “comprise”, and the like whenever used are intended to be non-restricting descriptive language in that they broadly include elements/components recited after such terms, in addition to other components not explicitly recited.
- reference to a “one” feature is also intended to be a reference to “at least one” of that feature.
- Terms such as “consisting”, “consist”, and the like may in the appropriate context, be considered as a subset of terms such as “comprising”, “comprise”, and the like.
- the individual numerical values within the range also include integers, fractions and decimals. Furthermore, whenever a range has been described, it is also intended that the range covers and teaches values of up to 2 additional decimal places or significant figures (where appropriate) from the shown numerical end points. For example, a description of a range of 1% to 5% is intended to have specifically disclosed the ranges 1.00% to 5.00% and also 1.0% to 5.0% and all their intermediate values (such as 1.01 %, 1.02% ... 4.98%, 4.99%, 5.00% and 1.1 %, 1 .2% ... 4.8%, 4.9%, 5.0% etc.,) spanning the ranges. The intention of the above specific disclosure is applicable to any depth/breadth of a range.
- the disclosure may have disclosed a method and/or process as a particular sequence of steps. However, unless otherwise required, it will be appreciated that the method or process should not be limited to the particular sequence of steps disclosed. Other sequences of steps may be possible. The particular order of the steps disclosed herein should not be construed as undue limitations. Unless otherwise required, a method and/or process disclosed herein should not be limited to the steps being carried out in the order written. The sequence of steps may be varied and still remain within the scope of the disclosure.
- an agent such as an antigen binding molecule/protein, that is capable of recognising/interacting with/binding to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and related products and methods are disclosed hereinafter.
- SARS-CoV-2 severe acute respiratory syndrome coronavirus 2
- SARS-CoV-2 Like two other pathogenic human respiratory coronaviruses (severe acute respiratory syndrome coronavirus [SARS-CoV] and Middle East respiratory syndrome coronavirus [MERS-CoV]), SARS-CoV-2, also known as COVID-19, can cause severe acute respiratory symptoms leading to mortality and morbidity. Common symptoms/features of a SARS-CoV-2 infection include fever, cough/dry cough, fatigue, myalgia, headache, dyspnea, sore throat, diarrhea, nausea/vomiting, loss of smell, loss of taste, abdominal pain, rhinorrhea and the like. In various embodiments, there is provided an agent that is capable of recognising/interacting with/binding to SARS-CoV-2.
- SARS-CoV-2 includes a virus having the sequence set forth in SEQ ID NO: 352, as well as variants/mutants thereof that share at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9%, at least about 99.91%, at least about 99.92%, at least about 99.93%, at least about 99.94%, at least about
- the agent may be in the form of a binding molecule, peptide (e.g., a polypeptide, an oligopeptide, a protein etc.), a peptidomimetic, an antigen binding protein, an antibody or fragments thereof (e.g., antigen binding fragments), derivatives thereof, combinations thereof, or the like.
- a derivative of a molecule is structurally related to the molecule.
- the derivative may share a common structural feature, fundamental structure and/or underlying chemical basis with the molecule.
- a derivative is not limited to one produced or obtained from the molecule although it may be one produced or obtained from the molecule.
- the derivative is derivable, at least theoretically, from the molecule through modification of the molecule.
- a derivative of a molecule shares or at least retains to a certain extent a function, chemical property, biological property, chemical activity and/or biological activity associated with the molecule.
- a skilled person will be able to identify, on a case-by-case basis and upon reading of the disclosure, the common structural feature, fundamental structure and/or underlying chemical basis of the molecule that have to be maintained in the derivative to retain the function, chemical property, biological property, chemical activity, and/or biological activity.
- assays that can prove the retention of the function, chemical property, biological property, chemical activity, and/or biological activity.
- a binding assay such as ELISA (enzyme-linked immunosorbent assay) may be carried out to determine a binding property of a derivative of a molecule.
- the agent comprises an antigen-binding protein.
- An antigen-binding protein may include any protein construct that is capable of binding to SARS-CoV-2 antigen. Examples include, but are not limited to, antibodies and fragments thereof such as antigen binding fragments. Non-limiting examples of antigen binding fragments include one or more fragments or portions of an antibody that retain the ability to specifically bind to an antigen (e.g., SARS- CoV-2), or synthetic modifications of an antibody fragments that retain the desired binding ability to the antigen.
- antigen binding fragments include single domain antibodies, further engineered molecules (such as, but is not limited to diabodies, triabodies, tetrabodies, minibodies, and the like), Fab fragments, Fab' fragments, F(ab fragments, Fd fragments, Fv fragments, single-chain Fv (scFv) molecules, seFv molecules, scFv dimer, BsFv molecules, dsFv molecules, (dsFv)2 molecules, dsFv-dsFv' molecules, Fv fragments, dAb fragments, bispecific antibodies, ds diabodies, nanobodies, domain antibodies, bivalent domain antibodies, and minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g.
- antigen binding fragments may retain at least about one, at least about two, at least about three, at least about four, at least about five or at least about six of the CDR regions/sequences of the antibody.
- Antigen-binding proteins may be of different formats/types, such as IgG, IgE, IgM, IgD, IgA, and IgY formats/types. They may also be of different subclasses/isotypes, such as lgG1 , lgG2, lgG3, lgG4, lgA1 and lgA2 subclasses/isotypes.
- the antigen-binding protein comprises an IgG-like structure/format or full-length IgG-like structure/format.
- the antigen-binding protein comprises or consists of a Fab fragment.
- a Fab fragment may be composed of a light chain and the variable and first constant domains of a heavy chain.
- a Fab fragment can be the product of papain cleavage of an antibody.
- the antigen-binding protein may be recombinant, chimeric, humanised or fully human.
- the antigen-binding protein comprises a fully human antigen-binding protein.
- a fully human antigen-binding protein includes an antibody in which both the CDR and framework regions comprise human sequences.
- Methods for producing fully human antigen-binding proteins/antibodies are known in the art. For example, fully human antigen binding proteins/antibodies may be produced in transgenic non-human mammals, e.g., mice, that have been genetically engineered with the human immunoglobulin loci. For example, fully human antigen-binding proteins/antibodies may be produced by phage display technology.
- the antigen-binding protein may comprise monoclonal antigen-binding protein.
- the monoclonal antigen-binding protein may be part of a population of substantially homogenous antigen-binding proteins that recognise the same epitope and display substantially the same binding specificity for the same epitope.
- Monoclonal antigen-binding proteins are typically identical in their amino acid sequences.
- the preparation of monoclonal antigen-binding proteins is not limited to any particular methods, and may include the use of hybridoma, recombinant techniques, phage display techniques, combinations thereof or any other techniques that are capable of producing a substantially uniform population of antigen-binding proteins.
- the antigen-binding protein comprises an isolated antigen-binding protein.
- isolated as used herein in relation to an antigen-binding protein or other molecule refers to such molecule that has been sufficiently separated from the environment with which it would naturally be associated (e.g., tissue, body fluid, etc.).
- An “isolated” antigen binding protein or molecule is therefore distinguished from any such molecule that may be present in situ within a human or animal body or present in a sample derived from a human or animal body.
- the sequence of an “isolated” antigen binding protein may, however, correspond to sequences found in a human or animal body.
- isolated does not necessarily mean the exclusion of artificial or synthetic mixtures with other components, or the presence of impurities, for example, due to incomplete purification.
- isolated antigen-binding proteins or molecules are also meant to include those that are chemically synthesised or engineered.
- an isolated antigen-binding protein or molecule is obtained by removing or purifying it from its natural environment, by selection from an antibody source as a phage display library or a B-cell repertoire or by chemical synthesis.
- an isolated antigen-binding protein comprises a Fab fragment selected from a phage display library.
- an isolated antigen-binding protein is obtained by cloning Fab sequences (e.g., sequences of a Fab fragment selected from a phage display library) to become an IgG format.
- the antigen-binding protein or parts thereof is naturally occurring.
- the antigen-binding protein or parts thereof is derived or obtained from a non-immunized and/or a healthy source/donor.
- the antigen-binding protein or parts thereof is derived from a non-infected source/donor e.g., a non-SARS-CoV-2-infected source/donor.
- the source/donor may be a mammal, optionally a human.
- the antigen-binding protein or parts thereof is derived or obtained from a naive phage display library, optionally a nafve human phage display library.
- the antigen-binding protein is derived or obtained from a Fab phage display library, optionally a human Fab phage display library, further optionally a nafve human Fab phage display library.
- a variable region of the antigen-binding protein is not further modified/engineered after being obtained from a nafve human Fab phage display library.
- the agent is engineered, synthetic or non- naturally-occurring.
- the antigen-binding protein is capable of binding specifically to SARS-CoV-2. In various embodiments, the antigen-binding protein is specific to SARS-CoV-2. In various examples, the antigen-binding protein is capable of binding to SARS-CoV-2 or portions thereof in a dose-dependent manner. In various embodiments, the antigen-binding protein does not substantially bind to or cross-react with a virus of another family that is not the coronavirus family.
- the antigen-binding protein is capable of binding to pseudotyped SARS-CoV-2.
- the antigen-binding protein is capable of binding to live SARS-CoV-2.
- a “live” SARS-CoV-2 may comprise a wild-type envelope protein.
- a “pseudotyped” virus may comprise a recombinant virus having a different envelope protein from the wild-type envelope protein.
- the spike protein of a pseudotyped virus may contain only 1254 amino acid residues (instead of 1273 amino acid residues) without the last 19 amino acid residues of the C-terminal region.
- the antigen-binding protein is capable of binding to live or pseudotyped SARS-CoV- 2 in solution and/or on surface.
- the antigen-binding protein is capable of recognising/interacting with/binding to portions of SARS-CoV-2 that mediate virus binding to or entry into a potential host cell.
- the antigen binding protein is capable of recognising/interacting with/binding to an envelope protein of SARS-CoV-2 or portions thereof.
- the antigen binding protein is capable of recognising/interacting with/binding to the spike protein (or spike glycoprotein) of SARS-CoV-2, e.g., a spike protein of SEQ ID NO: 353, or portions thereof.
- the antigen-binding protein is capable of recognising/interacting with/binding to the amino-terminal region or an amino-terminal unit of SARS-CoV-2 spike protein or portions thereof. In various embodiments, the antigen-binding protein is capable of recognising/interacting with/binding to the amino-terminal S1 subunit of SARS- CoV-2 spike protein or portions thereof. In various embodiments, the antigen binding protein is capable of recognising/interacting with/binding to the receptor binding domain (RBD) of SARS-CoV-2 spike protein, e.g., a RBD of SEQ ID NO: 354, or portions thereof. In various embodiments, the antigen-binding protein is capable of recognising/interacting with/binding to an epitope or an antigen on SARS-CoV-2.
- RBD receptor binding domain
- the antigen-binding protein has an ECso value (e.g., an average ECso value) of no more than about 100 nM, no more than about 90 nM, no more than about 80 nM, no more than about 70 nM, no more than about 60 nM, no more than about 50 nM, no more than about 40 nM, no more than about 30 nM, no more than about 20 nM, no more than about 19 nM, no more than about 18 nM, no more than about 17 nM, no more than about 16 nM, no more than about 15 nM, no more than about 14 nM, no more than about 13 nM, no more than about 12 nM, no more than about 11 nM, no more than about 10 nM, no more than about 9.5 nM, no more than about 9 nM, no more than about 8.5 nM, no more than about 8 nM, no more than about 7.5 nM, no more
- the antigen-binding protein has an EC50 value (e.g., an average EC50 value) of no more than about 80 nM, optionally no more than about 12 nM, further optionally no more than about 6 nM, further optionally no more than about 3 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 0.1 nM for SARS-CoV-2 e.g., a RBD of SARS-CoV-2.
- an EC50 value e.g., an average EC50 value
- the antigen-binding protein has an EC50 value (e.g., an average EC50 value) of from about 0.05 nM to about 80 nM, from about 0.05 nM to about 12 nM, from about 0.05 nM to about 10 nM, from about 0.05 nM to about 5 nM, from about 0.05 nM to about 2 nM, from about 0.05 nM to about 1 nM, from about 0.05 nM to about 0.5 nM, from about 0.05 nM to about 0.2 nM or from about 0.05 nM to about 0.1 nM for SARS-CoV-2 e.g., a RBD of SARS-CoV-2.
- SARS-CoV-2 e.g., a RBD of SARS-CoV-2.
- the antigen-binding protein is capable of blocking/interfering/inhibiting/hindering/disrupting one or more of the following function(s) of SARS-CoV-2 spike protein: (i) host cell attachment; (ii) receptor binding; and (iii) mediating host cell membrane and viral membrane fusion.
- the antigen-binding protein is capable of blocking/interfering/inhibiting/hindering/disrupting the attachment of SARS-CoV- 2 to a host cell.
- the antigen-binding protein is capable of blocking/interfering/inhibiting/hindering/disrupting the binding of SARS-CoV-2 to a cell entry receptor.
- the cell entry receptor comprises angiotensin converting enzyme II (ACE2).
- ACE2 angiotensin converting enzyme II
- the cell entry receptor comprises human ACE2.
- the antigen-binding protein has a potent blocking/interfering/inhibiting/hindering/disrupting action against the binding of SARS-CoV-2 to the cell entry receptor.
- the associated EC50 value of the antigen-binding protein is from about 0.1 nM to about 360 nM, from about 0.3 nM to about 100 nM, from about 0.3 nM to about 50 nM, from about 0.3 nM to about 25 nM, from about 0.3nM to about 2.5 nM, from about 0.3 nM to about 2.0nM or from about 0.3 nM to about 1 .5 nM.
- the associated EC50 value of the antigen-binding protein is no more than about 2.5 nM, no more than about 2.0 nM, or no more than about 1 .5 nM.
- the antigen-binding protein is capable of neutralising or mediating (or initiating) the neutralisation of SARS-CoV-2.
- the neutralizing antigen-binding protein may substantially prevent, block, inhibit or hinder the binding or association of SARS-CoV-2 to a cell (e.g., the binding or association of a RBD of a SARS-CoV-2 to a cell entry receptor ACE2 of a cell), thereby interrupting a biological response that would otherwise result from the interaction of SARS-CoV-2 with the cell.
- the neutralizing antigen-binding protein may substantially prevent, block, inhibit or hinder SARS-CoV-2 from infecting a cell.
- the antigen-binding protein is capable of substantially neutralising or mediating (or initiating) the neutralisation of SARS-CoV-2 in a dose-dependent manner.
- the antigen-binding protein is capable of substantially neutralising or mediating the neutralisation of SARS- CoV-2 entry into cells, optionally human cells, further optionally ACE2 expressing/presenting cells.
- the antigen binding protein comprises a SARS-CoV-2 neutralising antigen binding protein.
- the antigen-binding protein has a potent neutralising action against cell entry of SARS-CoV-2.
- the associated ECso value of the antigen-binding protein is from about 5 ng/ml to about 17000 ng/ml, from about 6 ng/ml to about 1000 ng/ml, from about 6 ng/ml to about 100 ng/ml, from about 6 ng/ml to about 60 ng/ml, from about 6 ng/ml to about 30 ng/ml or from about 6 ng/ml to about 12 ng/ml.
- the associated ECso value of the antigen-binding protein is no more than about 60 ng/ml, no more than about 30 ng/ml, or no more than about 12 ng/ml.
- the antigen-binding protein has an ICso value (e.g., an average ICso value) of no more than about 360 nM, no more than about 350 nM, no more than about 300 nM, no more than about 200 nM, no more than about 100 nM, no more than about 90 nM, no more than about 80 nM, no more than about 70 nM, no more than about 60 nM, no more than about 50 nM, no more than about 40 nM, no more than about 30 nM, no more than about 20 nM, no more than about 10 nM, no more than about 9.5 nM, no more than about 9 nM, no more than about 8.5 nM, no more than about 8 nM, no more than about 7.5 nM, no more than about 7 nM, no more than about 6.5 nM, no more than about 6 nM, no more than about 5.5 nM, no more than about 5 nM,
- the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of no more than about 360 nM, optionally no more than about 180 nM, further optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 5 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM or further optionally no more than about 0.5 nM for an inhibition of binding between SARS-CoV-2 and a cell, optionally for an inhibition of binding between a spike protein of SARS-CoV-2 and an ACE2 of a cell.
- an IC50 value e.g., an average IC50 value
- the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of from about 0.05 nM to about 360 nM, from about 0.05 nM to about 200 nM, from about 0.1 nM to about 100 nM, from about 0.1 nM to about 50 nM, from about 0.1 nM to about 10 nM, from about 0.1 nM to about 3 nM, from about 0.1 nM to about 1 .2 nM or from about 0.1 nM to about 0.5 nM for an inhibition of binding between SARS-CoV-2 and a cell, optionally for an inhibition of binding between a spike protein of SARS-CoV-2 and an ACE2 of a cell.
- an IC50 value e.g., an average IC50 value
- the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of no more than about 20,000 ng/ml, no more than about 10,000 ng/ml, no more than about 5000 ng/ml, no more than about 4000 ng/ml, no more than about 3000 ng/ml, no more than about 2000 ng/ml, no more than about 1000 ng/ml, no more than about 900 ng/ml, no more than about 800 ng/ml, no more than about 700 ng/ml, no more than about 600 ng/ml, no more than about 500 ng/ml, no more than about 450 ng/ml, no more than about 400 ng/ml, no more than about 350 ng/ml, no more than about 300 ng/ml, no more than about 250 ng/ml, no more than about 200 ng/ml, no more than about 150 ng/ml, no more than about 100 ng/
- the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of no more than about 20,000 ng/ml, optionally no more than about 10,000 ng/ml, further optionally no more than about 5000 ng/ml, further optionally no more than about 2500 ng/ml, further optionally no more than about 1000 ng/ml, further optionally no more than about 500 ng/ml, further optionally no more than about 250 ng/ml, further optionally no more than about 100 ng/ml, further optionally no more than about 50 ng/ml, further optionally no more than about 25 ng/ml or further optionally no more than about 10 ng/ml for an inhibition of SARS-CoV-2 entry into a cell, e.g., pseudotyped SARS-CoV-2 entry into an ACE2-expressing cell.
- IC50 value e.g., an average IC50 value
- the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of from about 5 ng/ml to about 20,000 ng/ml, from about 5 ng/ml to about 10,000 ng/ml, from about 5 ng/ml to about 1000 ng/ml, from about 5 ng/ml to about 500 ng/ml, from about 5 ng/ml to about 100 ng/ml, from about 5 ng/ml to about 50 ng/ml, from about 5 ng/ml to about 25 ng/ml or from about 5 ng/ml to about 10 ng/ml for an inhibition of SARS-CoV-2 entry into a cell, e.g., pseudotyped SARS-CoV-2 entry into an ACE2-expressing cell.
- IC50 value e.g., an average IC50 value
- the antigen-binding protein has a KD value (e.g., an average KD value) of no more than about 150 nM, no more than about 100 nM, no more than about 50 nM, no more than about 10 nM, no more than about 9 nM, no more than about 8 nM, no more than about 7 nM, no more than about
- KD value e.g., an average KD value
- nM 0.5 nM, no more than about 0.45 nM, no more than about 0.4 nM, no more than about 0.35 nM, no more than about 0.3 nM, no more than about 0.25 nM, no more than about 0.2 nM, no more than about 0.15 nM, no more than about 0.1 nM, no more than about 50 pM, no more than about 10 pM, no more than about 5 pM or no more than about 1 pM for SARS-CoV-2, e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry.
- SARS-CoV-2 e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry.
- the antigen binding protein has a KD value (e.g., an average KD value) of no more than about 130 nM, optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 1 pM for SARS-CoV-2, e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry.
- KD value e.g., an average KD value
- the antigen-binding protein has a KD value (e.g., an average KD value) of less than about 1 x 10 12 M for SARS-CoV-2, e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry.
- KD value e.g., an average KD value
- the antigen-binding protein is capable of inhibiting virus-cell fusion and/or cell-cell fusion mediated by SARS-CoV-2.
- the antigen-binding protein does not bind to, interact with or cross-react with other human coronaviruses (other than SARS-CoV-2). In some embodiments, the antigen-binding protein does not cross-react with severe acute respiratory syndrome coronavirus (SARS-CoV).
- SARS-CoV severe acute respiratory syndrome coronavirus
- embodiments of the antigen-binding protein have high specificity for SARS-CoV- 2.
- the antigen-binding protein is capable of binding to or interacting with other human coronaviruses (other than SARS-CoV-2). In some embodiments, the antigen-binding protein is capable of binding to or interacting with SARS-CoV in addition to SARS-CoV-2. Thus, embodiments of the antigen binding protein are capable of binding to an epitope that is conserved between SARS-CoV and SARS-CoV-2. Embodiments of the antigen-binding protein may be useful for detecting, binding to or neutralising two or more human coronaviruses such as SARS-CoV and SARS-CoV-2. In some embodiments, the antigen-binding protein binds to SARS-CoV-2 more strongly than SARS-CoV.
- the antigen-binding protein has a higher affinity for SARS- CoV-2 than SARS-CoV. In some embodiments, the antigen-binding protein neutralises SARS-CoV-2 by a greater extent as compared to SARS-CoV.
- the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising one or more mutations. In various embodiments, the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising no more than about 50, no more than about 40, no more than about 30, no more than about 20, no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s).
- the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising a mutated spike protein, for example, a mutated spike protein comprising no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s).
- a mutated spike protein comprising no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s).
- the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising a mutated RBD, for example, a mutated RBD comprising no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s).ln various embodiments the antigen-binding protein is capable of binding to and/or neutralizing at least about two, at least about three, at least about four, at least about five, at least about six, at least about seven or more of the following SARS-CoV-2: a SARS-CoV-2 comprising a wild-type spike protein (e.g., a spike protein of SEQ ID NO: 353) or a wild-type RBD (e.g., a RBD of SEQ ID NO: 354), a SARS-CoV-2 comprising a mutated spike protein with
- N439K, V483A, G476S, S494P, V483I, L455I/F456V and D614G mutations are all mutations occurring in the spike protein of SARS-CoV- 2, with the first six of them (i.e., N439K, V483A, G476S, S494P, V483I and L455I/F456V) occurring in the RBD and D614G occurring outside the RBD.
- the antigen-binding protein comprises an amino acid sequence corresponding to a sequence in Tables 1 -4. In various embodiments, the antigen-binding protein comprises an amino acid sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8% or at least about 99.9% with a sequence in Tables 1 -4.
- the antigen-binding protein comprises an amino acid sequence that differs by about one, about two, about three, about four, about five, about six, about seven, about eight, about nine, about ten or more amino acids with a sequence in Tables 1 -4.
- the antigen-binding protein comprises one or more amino acid region or complementarity determining region corresponding to a CDR identified in Tables 2 and 4. In various embodiments, the antigen-binding protein comprises one or more amino acid region or complementarity determining region sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence similarity/identity with a CDR or a combination of about two, about three, about four, about five or about six CDRs identified in Tables 2 and 4.
- the antigen-binding protein comprises an amino acid region or CDR that differs by about one, about two, about three, about four, about five, about six, about seven, about eight, about nine, about ten or more amino acids with a CDR or a combination of about two, about three, about four, about five or about six CDRs identified in Tables 2 and 4.
- the antigen-binding protein comprises an amino acid region, a light chain variable domain sequence and/or a heavy chain variable domain sequence corresponding to a light chain variable domain sequence and/or a heavy chain variable domain sequence identified in Tables 1 and 3.
- the antigen-binding protein comprises an amino acid region or a light chain variable domain and/or a heavy chain variable domain sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8% or at least about 99.9% with a light
- the antigen-binding protein comprises an amino acid region or a light chain variable domain and/or a heavy chain variable domain that differs by about one, about two, about three, about four, about five, about six, about seven, about eight, about nine, about ten or more amino acids with a light chain variable domain and/or a heavy chain variable domain identified in Tables 1 and 3.
- the antigen-binding protein comprises one of more of the following: a light-chain CDR1 selected from the group consisting of: SEQ ID NO: 28- 50 and conservative sequence variants thereof; a light-chain CDR2 selected from the group consisting of: SEQ ID NO: 51 - 64 and conservative sequence variants thereof; a light-chain CDR3 selected from the group consisting of: SEQ ID NO: 65- 90 and conservative sequence variants thereof; a heavy-chain CDR 1 selected from the group consisting of: SEQ ID NO: 117-132 and conservative sequence variants thereof; a heavy-chain CDR 2 selected from the group consisting of: SEQ ID NO: 133-148 and conservative sequence variants thereof; and a heavy-chain CDR 3 selected from the group consisting of: SEQ ID NO: 149-173 and conservative sequence variants thereof.
- the antigen-binding protein comprises the following CDR sequences:
- light-chain CDR1 SGSIASHY (SEQ ID NO: 28)
- light-chain CDR2 EDN (SEQ ID NO: 51 )
- light-chain CDR3 QSYGSGFVV (SEQ ID NO: 65)
- heavy-chain CDR1 GFTFSSYA (SEQ ID NO: 117)
- heavy-chain CDR2 ISGSGGST (SEQ ID NO: 133)
- heavy-chain CDR3 AKDYFRWL (SEQ ID NO: 149);
- light-chain CDR1 QSLLYSNGYNY (SEQ ID NO: 31 ) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQALQTPYT (SEQ ID NO: 68) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARDNPGYSSSWSPNWFDP (SEQ ID NO: 152);
- 1C2 light-chain CDR1 QSLLHSNGFTY (SEQ ID NO: 32) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQGIQTPLT (SEQ ID NO: 69) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSSAGTFYFDY (SEQ ID NO: 153);
- light-chain CDR1 SGSIASYY (SEQ ID NO: 33) light-chain CDR2: EDD (SEQ ID NO: 54) light-chain CDR3: QSFDSSIQHVV (SEQ ID NO: 70) heavy-chain CDR1 : GYPFSTYY (SEQ ID NO: 121) heavy-chain CDR2: IDPSGGST (SEQ ID NO: 137) heavy-chain CDR3: ARVAEQHLDYYFDY (SEQ ID NO: 154);
- D12 light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDGSAWV (SEQ ID NO: 71) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARLEWLRGAFDI (SEQ ID NO: 155);
- E5 light-chain CDR1 SGRIASNY (SEQ ID NO: 35) light-chain CDR2: EDT (SEQ ID NO: 55) light-chain CDR3: QSFDSGNQRVV (SEQ ID NO: 72) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARALVGKWLLLRGFDY (SEQ ID NO: 156);
- F4 light-chain CDR1 SSNIGRNF (SEQ ID NO: 36) light-chain CDR2: RNN (SEQ ID NO: 56) light-chain CDR3: QSYDNSLFVV (SEQ ID NO: 73) heavy-chain CDR1 : GDSVSSNSAA (SEQ ID NO: 122) heavy-chain CDR2: TYYRSKWYN (SEQ ID NO: 139) heavy-chain CDR3: AREKGIEGPAFDP (SEQ ID NO: 157);
- light-chain CDR1 SGSIASNY (SEQ ID NO: 34)
- light-chain CDR2 EDN (SEQ ID NO: 51)
- light-chain CDR3 QSYDENIRV (SEQ ID NO: 74)
- heavy-chain CDR1 GGSFSGYY (SEQ ID NO: 123)
- heavy-chain CDR2 INHSGST (SEQ ID NO: 140)
- heavy-chain CDR3 ARGRWLRGAFDI (SEQ ID NO: 158);
- 2C10 light-chain CDR1 SSTIGTNY (SEQ ID NO: 38) light-chain CDR2: DNY (SEQ ID NO: 58) light-chain CDR3: GTWDSRLSVGV (SEQ ID NO: 76) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INYRGNT (SEQ ID NO: 142) heavy-chain CDR3: ARWDGGNSVDH (SEQ ID NO: 160);
- 2C12 light-chain CDR1 QSINIY (SEQ ID NO: 39) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYITPQT (SEQ ID NO: 77) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARGGRFHYYYYAMDV (SEQ ID NO: 161);
- light-chain CDR1 QTVGGSY (SEQ ID NO: 37) light-chain CDR2: GAS (SEQ ID NO: 57) light-chain CDR3: QQYASSRT (SEQ ID NO: 75) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPILGIA (SEQ ID NO: 141) heavy-chain CDR3: ARHGGVGATTGYYYMDV (SEQ ID NO: 159);
- 2D3 light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ASSGWLRGAFDI (SEQ ID NO: 162);
- 2G7 light-chain CDR1 GGTIGSNY (SEQ ID NO: 40)
- light-chain CDR2 EDN (SEQ ID NO: 51 )
- light-chain CDR3 QSYDSSNRV (SEQ ID NO: 79)
- heavy-chain CDR1 GGSFSGYY (SEQ ID NO: 123)
- heavy-chain CDR2 INHSGST (SEQ ID NO: 140)
- heavy-chain CDR3: ARGKWLRGAFDI SEQ ID NO: 163);
- 2H4 light-chain CDR1 EFIGDD (SEQ ID NO: 41 ) light-chain CDR2: EAR (SEQ ID NO: 60) light-chain CDR3: LQYDSFPLT (SEQ ID NO: 80) heavy-chain CDR1 : GYTFSGYY (SEQ ID NO: 124) heavy-chain CDR2: VNPNSGGT (SEQ ID NO: 144) heavy-chain CDR3: ARGGRPGL P A AG Y I D Y (SEQ ID NO: 164);
- light-chain CDR1 SSDVGTYNY (SEQ ID NO: 42) light-chain CDR2: DVS (SEQ ID NO: 61 ) light-chain CDR3: ASFSSSSTLLV (SEQ ID NO: 81 ) heavy-chain CDR1 : GYTFSTYD (SEQ ID NO: 125) heavy-chain CDR2: ISTYNGDT (SEQ ID NO: 145) heavy-chain CDR3: ARGDSSVGYEYFQH (SEQ ID NO: 165);
- 3C5 light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGSLSGYY (SEQ ID NO: 126) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
- 3D11 light-chain CDR1 SGNIASNY (SEQ ID NO: 44)
- light-chain CDR2 EDN (SEQ ID NO: 51 )
- light-chain CDR3 QSYDNNIQV (SEQ ID NO: 83)
- heavy-chain CDR1 GGSFSGYY (SEQ ID NO: 123)
- heavy-chain CDR2 INHSGST (SEQ ID NO: 140)
- heavy-chain CDR3: ARRWWLRGAFDI SEQ ID NO: 166
- 3D2 light-chain CDR1 SGSIARNY (SEQ ID NO: 43) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDRTNKRV (SEQ ID NO: 82) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
- light-chain CDR1 SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNLQWV (SEQ ID NO: 84) heavy-chain CDR1 : GYTFTSYG (SEQ ID NO: 127) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARGIITMISDY (SEQ ID NO: 167);
- 3F1 light-chain CDR1 GGSIADNF (SEQ ID NO: 45) light-chain CDR2: DYS (SEQ ID NO: 62) light-chain CDR3: QSYDISNPV (SEQ ID NO: 85) heavy-chain CDR1 : GFTFSSYG (SEQ ID NO: 128) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARDGGGGMDV (SEQ ID NO: 168);
- 3F11 light-chain CDR1 HSDGRNY (SEQ ID NO: 46) light-chain CDR2: DTS (SEQ ID NO: 63) light-chain CDR3: HQFRRSVST (SEQ ID NO: 86) heavy-chain CDR1 : GYTFSSSG (SEQ ID NO: 129) heavy-chain CDR2: ISTYNGNT (SEQ ID NO: 146) heavy-chain CDR3: ATSIAVAGIDY (SEQ ID NO: 169);
- light-chain CDR1 SGNIASYY (SEQ ID NO: 48)
- light-chain CDR2 EDN (SEQ ID NO: 51)
- light-chain CDR3 QSYDRPNHVV (SEQ ID NO: 88)
- heavy-chain CDR1 GYTFTSYY (SEQ ID NO: 119)
- heavy-chain CDR2 INPSGGST (SEQ ID NO: 135)
- heavy-chain CDR3 ASSVVPAAIYDYYYGMDV (SEQ ID NO: 171);
- light-chain CDR1 TDSIASNY (SEQ ID NO: 47)
- light-chain CDR2 EDN (SEQ ID NO: 51)
- light-chain CDR3 QSYDGVNRGVI (SEQ ID NO: 87)
- heavy-chain CDR1 GFTFSNYG (SEQ ID NO: 130)
- heavy-chain CDR2 IWHDGTNK (SEQ ID NO: 147)
- heavy-chain CDR3 VRDQGAGVWNGYSY (SEQ ID NO: 170);
- light-chain CDR1 QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172);
- 6F8 light-chain CDR1 QTINNNY (SEQ ID NO: 50) light-chain CDR2: DAS (SEQ ID NO: 64) light-chain CDR3: QQYGSSPRT (SEQ ID NO: 90) heavy-chain CDR1 : GFTVSSNY (SEQ ID NO: 132) heavy-chain CDR2: IYSGGST (SEQ ID NO: 148) heavy-chain CDR3: ARDYGDYYFDY (SEQ ID NO: 173); or conservative sequence variants thereof (e.g., an antigen-binding protein comprising one CDR sequence that is a conservative sequence variant of one CDR sequence of 1A5 and five CDR sequences that are identical to the other five CDR sequences of 1A5, an antigen-binding protein comprising two CDR sequences that are conservative sequence variants of two CDR sequences of 1 B2 and three CDR sequences that are identical to the other three CDR sequences of 1 B2 etc.).
- an antigen-binding protein comprising one CDR
- antigen-binding protein comprises a light-chain variable region comprising any one of SEQ ID NO: 1 -27 or a sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence similarity/identity thereto; and/or a heavy-chain variable region comprising any one of SEQ ID NO: 91 -116 or a sequence sharing at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence similarity/identity thereto.
- the antigen-binding proteins in the combination comprises the CDR sequences of one or more of the following antigen-binding proteins: 1 A5, 1 A8, 1 B2, 1 B11 , 1 C2, 1 C3, 1 D12, 1 E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6 and 6F8.
- the antigen-binding proteins in the combination is selected from the group consisting of: 1A5, 1A8, 1 B2, 1 B11 , 1 C2, 1C3, 1 D12, 1 E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, 6F8 and combinations thereof.
- a first antigen-binding protein comprises the following CDR sequences:
- 5A6 light-chain CDR1 QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172) and a second antigen-binding protein comprises the following CDR sequences:
- 3D11 light-chain CDR1 SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
- compositions comprising the antigen-binding protein or the combination.
- the composition comprises a therapeutically effective amount of the antigen-binding protein or the combination e.g., for use in therapy.
- the composition further comprises a suitable carrier, adjuvant, diluent and/or excipient.
- the composition comprises a therapeutic composition.
- the composition comprises a pharmaceutical composition.
- the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier and/or pharmaceutically active compound.
- a pharmaceutical composition comprising the antigen-binding protein or the combination, and a pharmaceutically acceptable carrier.
- the antigen-binding protein or the combination for use in therapy. In various embodiments, there is provided the antigen-binding protein or the combination when used in therapy.
- the antigen-binding protein or the combination as a detecting agent, e.g., for detecting SARS-CoV-2. In various embodiments, there is provided the antigen-binding protein or the combination as a diagnostic agent e.g., for SARS-CoV-2.
- the antigen-binding protein or the combination for use in therapy, or as a diagnostic agent.
- a polynucleotide/nucleotide sequence encoding for the antigen-binding protein.
- the polynucleotide/nucleotide sequence may comprise DNA, cDNA and/or RNA.
- the polynucleotide/nucleotide sequence comprises one or more sequences set forth in Tables 5-8 or a sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence identity thereto.
- the polynucleotide/nucleotide sequence comprises one of more of SEQ ID NO: 174-351 or a sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence identity thereto.
- a vector comprising the polynucleotide/nucleotide sequence.
- the vector is selected from the group consisting of a phage, a plasmid, a viral particle, a baculovirus, a yeast plasmid, a lipid based vehicle, a polymer microsphere, a liposome, and a cell based vehicle, a colloidal gold particle, lipopolysaccharide, polypeptide, polysaccharide, a viral vehicle, an adenovirus, a retrovirus, a lentivirus, an adeno-associated viruses, a herpesvirus, a vaccinia virus, a foamy virus, a cytomegalovirus, a Semliki forest virus, a poxvirus, a pseudorabies virus, an RNA virus vector, a DNA virus vector and a vector derived from a combination of a plasmid and a phage DNA.
- the polynucleotide/nucleotide sequence is operatively linked to an expression control sequence(s) to direct peptide synthesis.
- the vector comprises one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells.
- a host cell transfected with or comprising the vector may comprise a bacterial cell, a yeast cell, an animal cell e.g., a mammalian cell and/or a plant cell.
- a bacteriophage a host cell or a non-human organism comprising the polynucleotide/nucleotide sequence.
- the non-human organism comprises a mammal.
- the non-human organism comprises a murine.
- a method of treating SARS-CoV- 2 infection in a subject comprising administering the antigen-binding protein, the combination or the composition (e.g., pharmaceutical composition) to the subject.
- the method comprises administering the subject a combination of two antigen-binding proteins, wherein a first antigen-binding protein comprises the following CDR sequences:
- 5A6 light-chain CDR1 QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172) and a second antigen-binding protein comprises the following CDR sequences:
- 3D11 light-chain CDR1 SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
- the method may be performed in vitro or ex vivo or in vivo.
- the antigen-binding protein e.g., pharmaceutical composition
- the combination or the composition e.g., pharmaceutical composition
- a method of detecting the presence of SARS-CoV-2 in a sample comprising: contacting the sample with the agent, e.g., the antigen-binding protein and detecting a binding event involving the agent, e.g., the antigen-binding protein.
- the detection of a binding event is indicative of the presence of SARS-CoV-2 in the sample.
- no detection of any binding events involving the agent, e.g., the antigen-binding protein is indicative of the absence of SARS-CoV-2 in the sample.
- the antigen binding protein may be labelled e.g., coupled to a label.
- the antigen-binding protein is labelled with horseradish peroxidase (FIRP) and tetramethylbenzidine (TMB) substrate may be added for visualization. It will be appreciated that other suitable labels and substrates may also be used for detecting a binding event.
- FIRP horseradish peroxidase
- TMB tetramethylbenzidine
- the antigen binding protein when used in detecting the presence of SARS-CoV-2 in a sample.
- the sample may be obtained from any source.
- the sample may be a biological sample, a pharmaceutical sample, an environmental sample, a food sample etc.
- the sample comprises a biological sample.
- biological samples include, but are not limited to blood, serum, plasma, sputum, saliva, lavage fluid (e.g.
- bronchial lavage fluid alveolar lavage fluid and bronchoalveolar lavage fluid
- sputum nasal fluid/swab/wash/aspirate
- anterior nares fluid/swab/wash/aspirate nasal mid turbinate fluid/swab/wash/aspirate
- pharyngeal fluid/swab/wash/aspirate nasopharyngeal fluid/swab/wash/aspirate
- oropharyngeal fluid/swab/wash/aspirate tissue biopsy e.g. lung biopsy, cerebrospinal fluid, urine, faeces, stool, anal swab, semen, sweat, tears, processed fractions thereof and the like.
- an agent optionally an antigen-binding protein, that is capable of recognising/interacting with/binding to SARS-CoV-2.
- the method comprises contacting a candidate compound with a SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/a SARS-CoV-2 spike protein RBD domain, wherein where the candidate compound binds to the SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain, the candidate agent is identified as a potential agent.
- the method comprises use of phage display technology to identify the agent.
- the method comprises an enzyme-linked immunosorbent assay (ELISA) method.
- the ELISA method comprises immobilizing SARS-CoV-2 spike protein RBD domain or antigen to a surface, adding a candidate compound to the surface under conditions suitable for binding, and determining a binding between the SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS- CoV-2 spike protein RBD domain or antigen and the candidate compound.
- the SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain or antigen is tagged (e.g., with biotin).
- the method comprises determining whether a candidate compound is capable of blocking a binding between SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain and ACE2 protein, wherein where the candidate compound is capable of blocking a binding between SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain and ACE2 protein, the candidate compound is identified as a potential agent.
- the method comprises providing a candidate compound that is bound to SARS-CoV- 2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain, and contacting the bound candidate compound with ACE2 protein, wherein where the bound candidate compound and/or the SARS-CoV-2/a SARS- CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain does not bind to the ACE2 protein, the candidate compound is identified as a potential agent.
- the method comprises determining whether a candidate compound is capable of neutralising cell entry of SARS-CoV-2 or potions thereof, wherein where the candidate compound is capable of neutralising cell entry of SARS-CoV-2 or portions thereof, the candidate compound is identified as a potential agent.
- the method comprises contacting SARS-CoV-2 (or a pseudotyped SARS-CoV-2) or potions thereof with ACE-expressing cells in the presence of the candidate compound, and determining an amount of entry of the SARS-CoV-2 or portions thereof into the ACE-expressing cells, wherein where the amount of entry of the SARS-CoV- 2 or portions thereof into the ACE-expressing cells is substantially low/reduced, the candidate compound is identified as a potential antigen-binding protein.
- the determining step comprises performing a luciferase assay.
- the candidate compound is pre-incubated with the
- SARS-CoV-2 or potions thereof before contact with the ACE-expressing cells are included in the SARS-CoV-2 or potions thereof before contact with the ACE-expressing cells.
- the method further comprises generating a library of sequences, optionally Fab sequences, to obtain the candidate compounds.
- the library comprises a phase display library.
- FIG. 1 shows binding ELISA of supernatants of 27 Fab clones to the antigen protein of biotinylated RBD-mFc.
- the 27 Fab clones were tested in a binding ELISA assay to assess their antigen binding to biotinylated RBD with a mouse Fc tag and detected by goat-anti-human Fab-HRP.
- the clones were tested with 70 mI of 0.2 pg/ml Biotinylated SARS-CoV-2 RBD_mFC, total 14 ng per well.
- FIG. 2 shows blocking of spike RBD protein binding to ACE2-His by supernatants of 27 Fab clones.
- the 27 Fab clones were tested in an ELISA assay to assess their potency in blocking spike protein binding to the recombinant ACE2 protein (ACE2 with His tag), with an irrelevant Fab clones used as a negative control.
- the clones were tested with 100 mI of 0.5 nM Biotinylated SARS-CoV-2 RBDjTiFC, total 0.05 pmol per well.
- FIG. 3 shows binding ELISA of 19 IgGs to the antigen protein of biotinylated RBD-mFc.
- the 19 antibody clones were tested in a binding ELISA assay to assess their antigen binding to biotinylated RBD with a mouse Fc tag, and detected by anti-human Fc-HRP, with an irrelevant lgG1 used as a negative control antibody.
- the clones were tested with 70 mI of 0.2 pg/ml Biotinylated SARS-CoV-2 RBDJTIFC, total 14 ng per well.
- FIG. 4 shows blocking of spike RBD protein binding to ACE2-Fc by 19 IgGs.
- the 19 antibody clones were tested in an ELISA assay to assess their potency in blocking spike protein binding to the recombinant ACE2 protein (ACE2 with human Fc tag), with an irrelevant IgG used as a negative control antibody.
- the clones were tested with 100 mI of 0.5 nM Biotinylated SARS- CoV-2 RBDJTIFC, total 0.05 pmol per well.
- FIG. 5 shows competition of RBD-binding between ACE2 and six IgGs by Bio-Layer Interferometry assay.
- a weak blocking IgG clone 1C3 black line
- FIG. 6 shows binding avidity of six IgGs to the RBD by Bio-Layer Interferometry assay.
- a range of IgG concentration from 12.5 nM to 0.39 nM (in 2-fold dilutions) are shown for each IgG.
- the sensorgrams are in grey lines and curve fittings are in black. Results shown are a representative of two independent experiments.
- FIG. 7 shows binding avidity of six IgG antibodies to SARS-CoV-2 (circle) and SARS-CoV (triangle) spike RBD proteins tested by ELISA.
- the 6 antibody IgG clones 1 F4, 2H4, 3D11 , 3F11 , 5A6 and 6F8 were tested in a binding ELISA assay to assess their antigen binding to biotinylated SARS-CoV-2 RBD with a mouse Fc tag as well as biotinylated SARS-CoV RBD with a His tag and detected by anti-human Fc-HRP.
- the antibodies were tested with 70 mI of 0.2 pg/ml Biotinylated SARS-CoV-2 RBDJTIFC, total 14 ng per well or 70 mI of 0.2 pg/ml Biotinylated SARS-CoV RBDJHis, total 14 ng per well.
- FIG. 8 shows epitope binding of 5A6 by Bio-Layer Interferometry (BLI) analysis. Buffer alone, an irrelevant antibody (AB) and 5A6 IgG were included as controls.
- BBI Bio-Layer Interferometry
- FIG. 9 shows binding affinity of five Fab clones to SARS-CoV-2 Spike RBD protein measured by Bio-Layer Interferometry.
- Fab binding to immobilized Fc- RBD was tested using a range of Fab concentrations from 100 nM to 3.125 nM (in 2-fold dilution).
- a representative set of measurements is shown with sensorgrams in grey and curve fittings in black.
- FIG. 10 shows binding of the 6 IgG antibodies (solid lines) and 5A6 Fab (dashed line,) to the purified SARS-CoV-2 pseudovirus.
- the antibodies were tested with 70 mI of 1 pg/ml of pseudoviral particles, total 70 ng per well.
- FIG. 11 shows SARS-CoV-2 pseudotyped virus neutralization assay by 19 IgGs. The 19 antibody clones were tested in a pseudotyped virus neutralization assay to assess their potency in neutralizing pseudotyped virus entry to ACE2 expression cells.
- FIG. 12 shows neutralization of the SARS-CoV-2 and SARS-CoV pseudotyped viruses by anti-SARS-CoV-2 spike RBD IgG and Fab antibodies.
- A Infection of CHO-ACE2 cells by SARS-CoV-2 pseudovirus were determined in the presence of 1 F4, 2H4, 3D11 , 3F11, 5A6 and 6F8 IgG antibodies.
- FIG. 13 shows binding avidity of 5A6 IgG (A) and 3D11 IgG (B) to the wildtype RBD and RBD mutants measured by biolayer interferometry.
- a range of 5A6 and 3D11 IgG concentrations from 12.5 nM to 0.39 nM (in 2-fold dilutions) are measured for each experiment.
- the sensorgrams are in grey lines and curve fittings are in black.
- FIG. 14 shows neutralization of the SARS-CoV-2 pseudovirus with either wildtype (WT) or RBD mutations by 5A6 (A) and 3D11 (B) IgGs. Data are presented as means ⁇ SEM in triplicates and are representative of two independent experiments.
- FIG. 15 shows anti-SARS-CoV-2 Spike RBD IgG antibodies affect trypsin induced cell syncytia formation.
- Vero E6 cells were transfected with furin recognition mutation of SARS-CoV-2 S-protein (R682RAR to A682AAR)-GFP. After 48 hours, the cell culture medium was changed to DMEM (no serum) and treated with or without antibodies and incubated for 1 hour at 37°C. The cells were then treated with or without trypsin 15 pg/ml for 2 hours at 37°C. Cells were fixed with 4%PFA and stained with DAPI.
- FIG. 16 shows the amino acid sequences of the antibodies as described herein. CDRs are shaded and identified consecutively as CDR1 (first shaded region), CDR2 (second shaded region), and CDR3 (third shaded region).
- FIG. 17 shows the nucleotide sequences of the antibodies as described herein.
- CDRs are shaded and identified consecutively as CDR1 (first shaded region), CDR2 (second shaded region), and CDR3 (third shaded region).
- SARS-CoV-2 A newly identified novel coronavirus (SARS-CoV-2) is causing pneumonia-associated respiratory syndrome across the world. Epidemiology, genomics, and pathogenesis of the SARS-CoV-2 show high homology with that of SARS-CoV. Current efforts are focusing on development of therapeutic agents for the treatment of coronavirus disease 2019 (COVID-19).
- SARS-CoV-2 also uses the angiotensin converting enzyme II (ACE2) as the cell entry receptor.
- ACE2 angiotensin converting enzyme II
- the spike protein contains an amino-terminal S1 unit and a carboxyl- terminal S2 subunit.
- the antibodies are 1A5, 1A8, 1 B2, 1 B11 , 1C2, 1C3, 1 D12, 1E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, and 6F8.
- the antibodies, derivatives thereof, or antigen-binding portions thereof disclosed herein are able to specifically target/bind SARS-CoV- 2 spike protein RBD both in solution and on the surface of the pseudotyped virus.
- Recombinant SARS-CoV-2 spike protein RBD domain with a mouse Fc tag (RBD-mFc) was used to isolate binders from a library of Fab sequences constructed by SlgN, using phage display technology.
- RBD-mFc mouse Fc tag
- 48 clones showing the positive Fab supernatant binding signals to RBD-mFc and potent antigen-receptor (RBD-ACE2) blocking were sequenced and 27 unique sequences were identified: 1 A5, 1 A8, 1 B2, 1 B11 , 1 C2, 1 C3, 1 D12, 1 E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, and 6F8 (FIG. 16, FIG. 17).
- ELISA and antigen-receptor blocking ELISA using Fab supernatants for the 27 unique clones were shown in FIG. 1 and FIG. 2, respectively.
- 19 clones were selected and cloned into lgG1 format for further characterisation: 1A8, 1 B11 , 1C2, 1C3, 1 E5, 1 F4, 2C12, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, and 6F8.
- the 19 antibody clones were tested in ELISA against biotinylated recombinant SARS-CoV-2 spike protein RBD-mFc to assess binding avidity for the target, with an irrelevant lgG1 used as a negative control antibody.
- neutravidin was coated at 5pg/ml onto the 96-well ELISA plates in coating buffer overnight at 4°C. After blocking with casein for 2 hours, biotinylated antigen of spike protein RBD-mFc at 0.2 pg/ml was added to the plates and captured by neutravidin during 1-hour incubation at room temperature.
- the present inventors also produced Fab antibodies for clone 1 F4, 2H4, 3D11 , 3F11 and 5A6 and measured their binding affinity to SARS-CoV-2 spike RBD protein by BLI (FIG. 9).
- a comparison of KD of Fab binding (affinity) and apparent KD of IgG binding (avidity) was also compared and shown in Table 9.
- KD of Fab based on 1 :1 Langmuir fitting and apparent KD of IgG based on 1 :2 bivalent analyte fitting of BLI sensorgrams for immobilized Fc-RBD Values are the Mean ⁇ SD of two independent experiments
- the SARS-CoV-2 pseudotyped virus was purified by gradient centrifugation and immobilized the viral particles on the ELISA plates to test if the antibodies as disclosed herein also bind efficiently to the natural RBD displayed on the viral surface.
- Concentration-dependent binding curves of the 5 IgG clones (1 F4, 2H4, 3D11 , 3F11 , and 5A6) revealed that 5A6 was packed on the viral surface at a much higher density than the other 4 antibodies and 5A6 Fab antibody, as shown by the higher optical signal (FIG. 10).
- the present disclosure tested the function of the 19 IgG antibody clones in neutralizing a pseudotyped virus particles expressing the spike protein of SARS-CoV-2. Briefly, serially diluted IgG antibodies were pre-incubated with an equal volume of pseudotyped virus (12 ng equivalent of p24) for 1 hour before the mixture was added to the monolayer of pre-seeded CHO-ACE2 cells. After 1 hour of pseudotyped virus infection, culture medium was topped up for further incubation of cells for another 48 hours. Then the cells were washed with PBS and lysed in Passive Lysis Buffer and the luciferase assay was performed using the Luciferase Assay System (Promega).
- the luciferase reading corresponds to the amount of entry of pseudotyped virus particles into the susceptible cells.
- the results showed that clone 1 F4, 5A6, 3F11 , 2H4 could potently neutralize the cell entry of pseudotyped virus (FIG. 11).
- a neutralization assay using both SARS-CoV-2 and SARS-CoV pseudotyped virus was performed with the 6 IgG clones 1 F4, 2H4, 3D11 , 3F11 , 5A6 and 6F8. All 6 antibodies showed dose- response neutralization (FIG. 12A).
- 5A6 IgG blocks cell-cell fusion
Abstract
The present invention relates to provide antigen-binding proteins that are capable of binding to and/or neutralizing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In one embodiment, the antigen-binding proteins are capable of binding to a spike protein of SARS-CoV-2 including receptor-binding domain (RBD) of SARS-CoV-2. The present invention also provides products comprising the same and methods of using the same for the treatment of SARS-CoV-2 infection or for detecting the presence of SARS-CoV-2 in a sample.
Description
ANTIGEN-BINDING PROTEIN TO SARS-CoV-2
TECHNICAL FIELD The present disclosure relates broadly to antigen-binding protein capable of binding to SARS-CoV-2, and related products and methods.
BACKGROUND At the end of 2019, a novel coronavirus, designated as SARS-CoV-2, emerged in Wuhan, China. Like outbreaks caused by two other pathogenic human respiratory coronaviruses (severe acute respiratory syndrome coronavirus [SARS-CoV] and Middle East respiratory syndrome coronavirus [MERS-CoV]), SARS-CoV-2, also known as COVID-19, causes acute respiratory disease that is often severe. As of March 2021 , nearly 120 million individuals worldwide have been confirmed to have COVID-19 infection with more than 2 million fatalities occurred. Given the SARS-CoV outbreak in 2002 and the MERS- CoV outbreak in 2012, SARS-CoV-2 is the third coronavirus to emerge in the human population in the past two decades — an emergence that has put global public health institutions on high alert.
Although vaccine is an important tool against the virus, detecting agents and therapeutic agents (e.g., virus neutralizing antibodies) may enhance detection, and also provide immediate protection and effective treatment which are urgently needed. Thus, there is a need to provide an agent, such as an antigen-binding protein, that is capable of binding and/or neutralising SARS-CoV- 2, and related products and methods.
SUMMARY In one aspect, there is provided an isolated antigen-binding protein that is capable of binding to and/or neutralizing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
In one embodiment, the antigen-binding protein is capable of binding to a spike protein of SARS-CoV-2, optionally a receptor-binding domain (RBD) of SARS-CoV-2.
In one embodiment, the antigen-binding protein comprises one or more of the following properties:
- an EC50 value of no more than about 80 nM, optionally no more than about 12 nM, further optionally no more than about 6 nM, further optionally no more than about 3 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 0.1 nM for SARS-CoV-2;
- an IC50 value of no more than about 360 nM, optionally no more than about 180 nM, further optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 5 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM or further optionally no more than about 0.5 nM for an inhibition of binding between SARS-CoV-2 and a cell;
- an IC50 value of no more than about 20,000 ng/ml, optionally no more than about 10,000 ng/ml, further optionally no more than about 5000 ng/ml, further optionally no more than about 2500 ng/ml, further optionally no more than about 1000 ng/ml, further optionally no more than about 500 ng/ml, further optionally no more than about 250 ng/ml, further optionally no more than about 100 ng/ml, further optionally no more than about 50 ng/ml, further optionally no more than about 25 ng/ml or further optionally no more than about 10 ng/ml for an inhibition of SARS-CoV-2 entry into a cell; and/or
- a KD value of no more than about 130 nM, optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 1 pM for SARS-CoV-2.
In one embodiment, the antigen-binding protein does not cross-react with severe acute respiratory syndrome coronavirus (SARS-CoV).
In one embodiment, the antigen-binding protein is capable of binding to SARS-CoV.
In one embodiment, the antigen-binding protein comprises a monoclonal antigen-binding protein.
In one embodiment, the antigen-binding protein comprises a fully human antigen-binding protein.
In one embodiment, the antigen-binding protein comprises an IgG-like structure.
In one embodiment, the antigen-binding protein comprises a Fab fragment.
In one embodiment, the antigen-binding protein is capable of binding to and/or neutralizing about two or more of the following SARS-CoV-2: a SARS- CoV-2 comprising a wild-type spike protein, a SARS-CoV-2 comprising a mutated spike protein with no more than about two mutations, a SARS-CoV-2 comprising a mutated spike protein with no more than about one mutation, a SARS-CoV-2 comprising a N439K mutation, a SARS-CoV-2 comprising a V483A mutation, a SARS-CoV-2 comprising a G476S mutation, a SARS-CoV-2 comprising a S494P mutation, a SARS-CoV-2 comprising a V483I mutation, a SARS-CoV-2 comprising L455I and F456V mutations and a SARS-CoV-2 comprising a D614G mutation.
In one embodiment, the antigen-binding protein is capable of inhibiting virus-cell fusion and/or cell-cell fusion mediated by SARS-CoV-2.
In one embodiment, the antigen-binding protein comprises: a light-chain complementarity determining region CDR1 selected from the group consisting of: SEQ ID NO: 28-50 and conservative sequence variants thereof; a light-chain CDR2 selected from the group consisting of: SEQ ID NO: 51 - 64 and conservative sequence variants thereof; a light-chain CDR3 selected from the group consisting of: SEQ ID NO: 65- 90 and conservative sequence variants thereof; a heavy-chain CDR 1 selected from the group consisting of: SEQ ID NO: 117-132 and conservative sequence variants thereof;
a heavy-chain CDR 2 selected from the group consisting of: SEQ ID NO: 133-148 and conservative sequence variants thereof; and a heavy-chain CDR 3 selected from the group consisting of: SEQ ID NO: 149-173 and conservative sequence variants thereof.
In one embodiment, the antigen-binding protein comprises the following CDR sequences:
1A5 light-chain CDR1 : SGSIASHY (SEQ ID NO: 28) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYGSGFVV (SEQ ID NO: 65) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISGSGGST (SEQ ID NO: 133) heavy-chain CDR3: AKDYFRWL (SEQ ID NO: 149);
1A8 light-chain CDR1 : QSVLYSSNNKNY (SEQ ID NO: 29) light-chain CDR2: WAS (SEQ ID NO: 52) light-chain CDR3: QQYYGTPYT (SEQ ID NO: 66) heavy-chain CDR1 : GVSISSRSDH (SEQ ID NO: 118) heavy-chain CDR2: ISYSGST (SEQ ID NO: 134) heavy-chain CDR3: ARLASLYSTFDI (SEQ ID NO: 150);
1 B2 light-chain CDR1 : GGRIATNY (SEQ ID NO: 30) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSHDTSRQAV (SEQ ID NO: 67) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ATGEMAGDFDY (SEQ ID NO: 151);
1 B11 light-chain CDR1 : QSLLYSNGYNY (SEQ ID NO: 31) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQALQTPYT (SEQ ID NO: 68) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARDNPGYSSSWSPNWFDP (SEQ ID NO: 152);
1 C2 light-chain CDR1 : QSLLHSNGFTY (SEQ ID NO: 32) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQGIQTPLT (SEQ ID NO: 69) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSSAGTFYFDY (SEQ ID NO: 153);
1C3 light-chain CDR1 : SGSIASYY (SEQ ID NO: 33) light-chain CDR2: EDD (SEQ ID NO: 54) light-chain CDR3: QSFDSSIQHVV (SEQ ID NO: 70) heavy-chain CDR1 : GYPFSTYY (SEQ ID NO: 121) heavy-chain CDR2: IDPSGGST (SEQ ID NO: 137) heavy-chain CDR3: ARVAEQHLDYYFDY (SEQ ID NO: 154);
1 D12 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDGSAWV (SEQ ID NO: 71) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARLEWLRGAFDI (SEQ ID NO: 155);
1 E5 light-chain CDR1 : SGRIASNY (SEQ ID NO: 35) light-chain CDR2: EDT (SEQ ID NO: 55) light-chain CDR3: QSFDSGNQRVV (SEQ ID NO: 72) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARALVGKWLLLRGFDY (SEQ ID NO: 156);
1 F4 light-chain CDR1 : SSNIGRNF (SEQ ID NO: 36) light-chain CDR2: RNN (SEQ ID NO: 56) light-chain CDR3: QSYDNSLFVV (SEQ ID NO: 73) heavy-chain CDR1 : GDSVSSNSAA (SEQ ID NO: 122) heavy-chain CDR2: TYYRSKWYN (SEQ ID NO: 139) heavy-chain CDR3: AREKGIEGPAFDP (SEQ ID NO: 157);
1 H7 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDENIRV (SEQ ID NO: 74) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGRWLRGAFDI (SEQ ID NO: 158);
2C9 light-chain CDR1 : QTVGGSY (SEQ ID NO: 37) light-chain CDR2: GAS (SEQ ID NO: 57) light-chain CDR3: QQYASSRT (SEQ ID NO: 75) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPILGIA (SEQ ID NO: 141) heavy-chain CDR3: ARHGGVGATTGYYYMDV (SEQ ID NO: 159);
2C10 light-chain CDR1 : SSTIGTNY (SEQ ID NO: 38) light-chain CDR2: DNY (SEQ ID NO: 58) light-chain CDR3: GTWDSRLSVGV (SEQ ID NO: 76) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INYRGNT (SEQ ID NO: 142) heavy-chain CDR3: ARWDGGNSVDH (SEQ ID NO: 160);
2C12 light-chain CDR1 : QSINIY (SEQ ID NO: 39) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYITPQT (SEQ ID NO: 77) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARGGRFHYYYYAMDV (SEQ ID NO: 161);
2D3 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ASSGWLRGAFDI (SEQ ID NO: 162);
2G7 light-chain CDR1 : GGTIGSNY (SEQ ID NO: 40) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNRV (SEQ ID NO: 79) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGKWLRGAFDI (SEQ ID NO: 163);
2H4 light-chain CDR1 : EFIGDD (SEQ ID NO: 41) light-chain CDR2: EAR (SEQ ID NO: 60) light-chain CDR3: LQYDSFPLT (SEQ ID NO: 80) heavy-chain CDR1 : GYTFSGYY (SEQ ID NO: 124) heavy-chain CDR2: VNPNSGGT (SEQ ID NO: 144) heavy-chain CDR3: ARGGRPGLPAAGYIDY (SEQ ID NO: 164);
3A11 light-chain CDR1 : SSDVGTYNY (SEQ ID NO: 42) light-chain CDR2: DVS (SEQ ID NO: 61) light-chain CDR3: ASFSSSSTLLV (SEQ ID NO: 81) heavy-chain CDR1 : GYTFSTYD (SEQ ID NO: 125) heavy-chain CDR2: ISTYNGDT (SEQ ID NO: 145) heavy-chain CDR3: ARGDSSVGYEYFQH (SEQ ID NO: 165);
3C5 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGSLSGYY (SEQ ID NO: 126) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3D2 light-chain CDR1 : SGSIARNY (SEQ ID NO: 43) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDRTNKRV (SEQ ID NO: 82) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3E9 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNLQWV (SEQ ID NO: 84) heavy-chain CDR1 : GYTFTSYG (SEQ ID NO: 127) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARGIITMISDY (SEQ ID NO: 167);
3F1 light-chain CDR1 : GGSIADNF (SEQ ID NO: 45) light-chain CDR2: DYS (SEQ ID NO: 62) light-chain CDR3: QSYDISNPV (SEQ ID NO: 85) heavy-chain CDR1 : GFTFSSYG (SEQ ID NO: 128) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARDGGGGMDV (SEQ ID NO: 168);
3F11 light-chain CDR1 : HSDGRNY (SEQ ID NO: 46) light-chain CDR2: DTS (SEQ ID NO: 63) light-chain CDR3: HQFRRSVST (SEQ ID NO: 86) heavy-chain CDR1 : GYTFSSSG (SEQ ID NO: 129) heavy-chain CDR2: ISTYNGNT (SEQ ID NO: 146) heavy-chain CDR3: ATSIAVAGIDY (SEQ ID NO: 169);
3H7 light-chain CDR1 : TDSIASNY (SEQ ID NO: 47) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDGVNRGVI (SEQ ID NO: 87) heavy-chain CDR1 : GFTFSNYG (SEQ ID NO: 130) heavy-chain CDR2: IWHDGTNK (SEQ ID NO: 147) heavy-chain CDR3: VRDQGAGVWNGYSY (SEQ ID NO: 170);
3H11 light-chain CDR1 : SGNIASYY (SEQ ID NO: 48) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDRPNHVV (SEQ ID NO: 88) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSVVPAAIYDYYYGMDV (SEQ ID NO: 171);
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172);
6F8 light-chain CDR1 : QTINNNY (SEQ ID NO: 50) light-chain CDR2: DAS (SEQ ID NO: 64) light-chain CDR3: QQYGSSPRT (SEQ ID NO: 90) heavy-chain CDR1 : GFTVSSNY (SEQ ID NO: 132) heavy-chain CDR2: IYSGGST (SEQ ID NO: 148) heavy-chain CDR3: ARDYGDYYFDY (SEQ ID NO: 173); or
conservative sequence variants thereof.
In one embodiment, the antigen-binding protein comprises a light-chain variable region comprising any one of SEQ ID NO: 1 -27 or a sequence sharing at least about 75% sequence similarity thereto; and/or a heavy-chain variable region comprising any one of SEQ ID NO: 91 -116 or a sequence sharing at least about 75% sequence similarity thereto.
In one aspect, there is provided a combination of two antigen-binding proteins, wherein a first antigen-binding protein comprises the following CDR sequences:
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172) and a second antigen-binding protein comprises the following CDR sequences:
3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
In one aspect, there is provided a pharmaceutical composition comprising embodiments of the antigen-binding protein or embodiments of the combination, and a pharmaceutically acceptable carrier.
In one aspect, there is provided embodiments of the antigen-binding protein, embodiments of the combination or embodiments of the pharmaceutical composition, for use in therapy, or as a diagnostic agent.
In one aspect, there is provided a nucleotide sequence encoding for embodiments of the antigen-binding protein or embodiments of the combination.
In one embodiment, the nucleotide sequence comprises one of more of SEQ ID NO: 174-351 or a sequence sharing at least about 75% sequence identity thereto.
In one aspect, there is provided a bacteriophage, a host cell or a non-human organism comprising embodiments of the nucleotide sequence.
In one aspect, there is provided a method of treating SARS-CoV-2 in a subject, the method comprising administering to the subject embodiments of the antigen-binding protein, embodiments of the combination or embodiments of the pharmaceutical composition.
In one embodiment, where the SARS-CoV-2 comprises a V483A mutation, the method comprises administering to the subject embodiments of the combination.
In one aspect, there is provided a method of detecting the presence of SARS-CoV-2 in a sample, the method comprising: contacting the sample with the embodiments of the antigen-binding protein; detecting a binding event involving the antigen-binding protein; wherein the detection of a binding event is indicative of the presence of SARS-CoV-2 in the sample.
DEFINITIONS
The term “treating", "treat" and “therapy”, and synonyms thereof as used herein refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) a medical
condition, which includes but is not limited to diseases (such as SARS-CoV-2 infections), symptoms and disorders. A medical condition also includes a body’s response to a disease or disorder, e.g., inflammation. Those in need of such treatment include those already with a medical condition as well as those prone to getting the medical condition or those in whom a medical condition is to be prevented.
As used herein, the term "therapeutically effective amount" will be an amount of an active agent that is capable of preventing, reversing or at least slowing down (lessening) a medical condition, such as SARS-CoV-2 infections Dosages and administration of agents, compounds, compositions and formulations of the present disclosure may be determined by one of ordinary skill in the art of clinical pharmacology or pharmacokinetics. See, for example, Mordenti and Rescigno, (1992) Pharmaceutical Research. 9:17-25; Morenti et al., (1991) Pharmaceutical Research. 8:1351 -1359; and Mordenti and Chappell, "The use of interspecies scaling in toxicokinetics" in Toxicokinetics and New Drug Development, Yacobi et al. (eds) (Pergamon Press: NY, 1989), pp. 42-96. An effective amount of the active agent of the present disclosure to be employed therapeutically will depend, for example, upon the therapeutic objectives, the route of administration, and the condition of the subject. Accordingly, it may be necessary for the therapist to titer the dosage and modify the route of administration as required to obtain the optimal therapeutic effect.
The term “subject” as used herein includes patients and non-patients. The term “patient” refers to individuals suffering or are likely to suffer from a medical condition such as a SARS-CoV-2 infection, while “non-patients” refer to individuals not suffering and are likely to not suffer from the medical condition. “Non-patients” include healthy individuals, non-diseased individuals and/or an individual free from the medical condition. The term “subject” includes humans and animals. Animals may include, but is not limited to, mammals (for example non-human primates, canine, murine and the like), and the like. “Murine” refers to any mammal from the family Muridae, such as mouse, rat, rabbit, and the like.
The term "micro" as used herein is to be interpreted broadly to include dimensions from about 1 micron to about 1000 microns.
The term "nano" as used herein is to be interpreted broadly to include dimensions less than about 1000 nm.
The term “particle” as used herein broadly refers to a discrete entity or a discrete body. The particle described herein can include an organic, an inorganic or a biological particle. The particle used described herein may also be a macro particle that is formed by an aggregate of a plurality of sub-particles or a fragment of a small object. The particle of the present disclosure may be spherical, substantially spherical, or non-spherical, such as irregularly shaped particles or ellipsoidally shaped particles. The term “size” when used to refer to the particle broadly refers to the largest dimension of the particle. For example, when the particle is substantially spherical, the term “size” can refer to the diameter of the particle; or when the particle is substantially non-spherical, the term “size” can refer to the largest length of the particle.
The terms "coupled" or "connected" as used in this description are intended to cover both directly connected or connected through one or more intermediate means, unless otherwise stated.
The term "associated with", used herein when referring to two elements refers to a broad relationship between the two elements. The relationship includes, but is not limited to a physical, a chemical or a biological relationship. For example, when element A is associated with element B, elements A and B may be directly or indirectly attached to each other or element A may contain element B or vice versa.
The term "adjacent" used herein when referring to two elements refers to one element being in close proximity to another element and may be but is not limited to the elements contacting each other or may further include the elements being separated by one or more further elements disposed therebetween.
The term "and/or", e.g., "X and/or Y" is understood to mean either "X and Y" or "X or Y" and should be taken to provide explicit support for both meanings or for either meaning.
Further, in the description herein, the word “substantially” whenever used is understood to include, but not restricted to, "entirely" or “completely” and the like. In addition, terms such as "comprising", "comprise", and the like whenever
used, are intended to be non-restricting descriptive language in that they broadly include elements/components recited after such terms, in addition to other components not explicitly recited. For example, when “comprising” is used, reference to a “one” feature is also intended to be a reference to “at least one” of that feature. Terms such as “consisting”, “consist”, and the like, may in the appropriate context, be considered as a subset of terms such as "comprising", "comprise", and the like. Therefore, in embodiments disclosed herein using the terms such as "comprising", "comprise", and the like, it will be appreciated that these embodiments provide teaching for corresponding embodiments using terms such as “consisting”, “consist”, and the like. Further, terms such as "about", "approximately" and the like whenever used, typically means a reasonable variation, for example a variation of +/- 5% of the disclosed value, or a variance of 4% of the disclosed value, or a variance of 3% of the disclosed value, a variance of 2% of the disclosed value or a variance of 1% of the disclosed value.
Furthermore, in the description herein, certain values may be disclosed in a range. The values showing the end points of a range are intended to illustrate a preferred range. Whenever a range has been described, it is intended that the range covers and teaches all possible sub-ranges as well as individual numerical values within that range. That is, the end points of a range should not be interpreted as inflexible limitations. For example, a description of a range of 1% to 5% is intended to have specifically disclosed sub-ranges 1% to 2%, 1% to 3%, 1% to 4%, 2% to 3% etc., as well as individually, values within that range such as 1%, 2%, 3%, 4% and 5%. It is to be appreciated that the individual numerical values within the range also include integers, fractions and decimals. Furthermore, whenever a range has been described, it is also intended that the range covers and teaches values of up to 2 additional decimal places or significant figures (where appropriate) from the shown numerical end points. For example, a description of a range of 1% to 5% is intended to have specifically disclosed the ranges 1.00% to 5.00% and also 1.0% to 5.0% and all their intermediate values (such as 1.01 %, 1.02% ... 4.98%, 4.99%, 5.00% and 1.1 %, 1 .2% ... 4.8%, 4.9%, 5.0% etc.,) spanning the ranges. The intention of the above specific disclosure is applicable to any depth/breadth of a range.
Additionally, when describing some embodiments, the disclosure may have disclosed a method and/or process as a particular sequence of steps. However, unless otherwise required, it will be appreciated that the method or process should not be limited to the particular sequence of steps disclosed. Other sequences of steps may be possible. The particular order of the steps disclosed herein should not be construed as undue limitations. Unless otherwise required, a method and/or process disclosed herein should not be limited to the steps being carried out in the order written. The sequence of steps may be varied and still remain within the scope of the disclosure. Furthermore, it will be appreciated that while the present disclosure provides embodiments having one or more of the features/characteristics discussed herein, one or more of these features/characteristics may also be disclaimed in other alternative embodiments and the present disclosure provides support for such disclaimers and these associated alternative embodiments.
DESCRIPTION OF EMBODIMENTS
Exemplary, non-limiting embodiments of an agent, such as an antigen binding molecule/protein, that is capable of recognising/interacting with/binding to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and related products and methods are disclosed hereinafter.
Like two other pathogenic human respiratory coronaviruses (severe acute respiratory syndrome coronavirus [SARS-CoV] and Middle East respiratory syndrome coronavirus [MERS-CoV]), SARS-CoV-2, also known as COVID-19, can cause severe acute respiratory symptoms leading to mortality and morbidity. Common symptoms/features of a SARS-CoV-2 infection include fever, cough/dry cough, fatigue, myalgia, headache, dyspnea, sore throat, diarrhea, nausea/vomiting, loss of smell, loss of taste, abdominal pain, rhinorrhea and the like. In various embodiments, there is provided an agent that is capable of recognising/interacting with/binding to SARS-CoV-2. The term “SARS-CoV-2” includes a virus having the sequence set forth in SEQ ID NO: 352, as well as
variants/mutants thereof that share at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9%, at least about 99.91%, at least about 99.92%, at least about 99.93%, at least about 99.94%, at least about 99.95%, at least about 99.96%, at least about 99.97%, at least about 99.98% or at least about 99.99% sequence identity with the sequence over its entire length. Advantageously, embodiments of the agent are useful as therapeutics and detecting agents for SARS-CoV-2.
The agent may be in the form of a binding molecule, peptide (e.g., a polypeptide, an oligopeptide, a protein etc.), a peptidomimetic, an antigen binding protein, an antibody or fragments thereof (e.g., antigen binding fragments), derivatives thereof, combinations thereof, or the like. In various embodiments, a derivative of a molecule is structurally related to the molecule. For example, the derivative may share a common structural feature, fundamental structure and/or underlying chemical basis with the molecule. A derivative is not limited to one produced or obtained from the molecule although it may be one produced or obtained from the molecule. In some embodiments, the derivative is derivable, at least theoretically, from the molecule through modification of the molecule. In some embodiments, a derivative of a molecule shares or at least retains to a certain extent a function, chemical property, biological property, chemical activity and/or biological activity associated with the molecule. A skilled person will be able to identify, on a case-by-case basis and upon reading of the disclosure, the common structural feature, fundamental structure and/or underlying chemical basis of the molecule that have to be maintained in the derivative to retain the function, chemical property, biological property, chemical activity, and/or biological activity. A skilled person will also be able to identify assays that can prove the retention of the function, chemical property, biological property,
chemical activity, and/or biological activity. For example, a binding assay such as ELISA (enzyme-linked immunosorbent assay) may be carried out to determine a binding property of a derivative of a molecule.
In one embodiment, the agent comprises an antigen-binding protein. An antigen-binding protein may include any protein construct that is capable of binding to SARS-CoV-2 antigen. Examples include, but are not limited to, antibodies and fragments thereof such as antigen binding fragments. Non-limiting examples of antigen binding fragments include one or more fragments or portions of an antibody that retain the ability to specifically bind to an antigen (e.g., SARS- CoV-2), or synthetic modifications of an antibody fragments that retain the desired binding ability to the antigen. In various embodiments, antigen binding fragments include single domain antibodies, further engineered molecules (such as, but is not limited to diabodies, triabodies, tetrabodies, minibodies, and the like), Fab fragments, Fab' fragments, F(ab fragments, Fd fragments, Fv fragments, single-chain Fv (scFv) molecules, seFv molecules, scFv dimer, BsFv molecules, dsFv molecules, (dsFv)2 molecules, dsFv-dsFv' molecules, Fv fragments, dAb fragments, bispecific antibodies, ds diabodies, nanobodies, domain antibodies, bivalent domain antibodies, and minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g. an isolated complementarity determining region (CDR)). In some embodiments, antigen binding fragments may retain at least about one, at least about two, at least about three, at least about four, at least about five or at least about six of the CDR regions/sequences of the antibody.
Antigen-binding proteins may be of different formats/types, such as IgG, IgE, IgM, IgD, IgA, and IgY formats/types. They may also be of different subclasses/isotypes, such as lgG1 , lgG2, lgG3, lgG4, lgA1 and lgA2 subclasses/isotypes. In some embodiments, the antigen-binding protein comprises an IgG-like structure/format or full-length IgG-like structure/format. In some embodiments, the antigen-binding protein comprises or consists of a Fab fragment. A Fab fragment may be composed of a light chain and the variable and first constant domains of a heavy chain. A Fab fragment can be the product of papain cleavage of an antibody.
The antigen-binding protein may be recombinant, chimeric, humanised or fully human. In various embodiments, the antigen-binding protein comprises a fully human antigen-binding protein. A fully human antigen-binding protein includes an antibody in which both the CDR and framework regions comprise human sequences. Methods for producing fully human antigen-binding proteins/antibodies are known in the art. For example, fully human antigen binding proteins/antibodies may be produced in transgenic non-human mammals, e.g., mice, that have been genetically engineered with the human immunoglobulin loci. For example, fully human antigen-binding proteins/antibodies may be produced by phage display technology.
In various embodiments, the antigen-binding protein may comprise monoclonal antigen-binding protein. The monoclonal antigen-binding protein may be part of a population of substantially homogenous antigen-binding proteins that recognise the same epitope and display substantially the same binding specificity for the same epitope. Monoclonal antigen-binding proteins are typically identical in their amino acid sequences. The preparation of monoclonal antigen-binding proteins is not limited to any particular methods, and may include the use of hybridoma, recombinant techniques, phage display techniques, combinations thereof or any other techniques that are capable of producing a substantially uniform population of antigen-binding proteins.
In some embodiments, the antigen-binding protein comprises an isolated antigen-binding protein. The term "isolated" as used herein in relation to an antigen-binding protein or other molecule refers to such molecule that has been sufficiently separated from the environment with which it would naturally be associated (e.g., tissue, body fluid, etc.). An “isolated” antigen binding protein or molecule is therefore distinguished from any such molecule that may be present in situ within a human or animal body or present in a sample derived from a human or animal body. The sequence of an “isolated” antigen binding protein may, however, correspond to sequences found in a human or animal body. The term “isolated” does not necessarily mean the exclusion of artificial or synthetic mixtures with other components, or the presence of impurities, for example, due to incomplete purification. In particular, isolated antigen-binding proteins or
molecules are also meant to include those that are chemically synthesised or engineered. In various embodiments, an isolated antigen-binding protein or molecule is obtained by removing or purifying it from its natural environment, by selection from an antibody source as a phage display library or a B-cell repertoire or by chemical synthesis. In some embodiments, an isolated antigen-binding protein comprises a Fab fragment selected from a phage display library. In some embodiments, an isolated antigen-binding protein is obtained by cloning Fab sequences (e.g., sequences of a Fab fragment selected from a phage display library) to become an IgG format. In some embodiments, the antigen-binding protein or parts thereof is naturally occurring. In some embodiments, the antigen-binding protein or parts thereof is derived or obtained from a non-immunized and/or a healthy source/donor. In some embodiments, the antigen-binding protein or parts thereof is derived from a non-infected source/donor e.g., a non-SARS-CoV-2-infected source/donor. The source/donor may be a mammal, optionally a human. In some embodiments, the antigen-binding protein or parts thereof is derived or obtained from a naive phage display library, optionally a nafve human phage display library. In some embodiments, the antigen-binding protein is derived or obtained from a Fab phage display library, optionally a human Fab phage display library, further optionally a nafve human Fab phage display library. In various embodiments, a variable region of the antigen-binding protein is not further modified/engineered after being obtained from a nafve human Fab phage display library. In some embodiments, the agent is engineered, synthetic or non- naturally-occurring. In various embodiments therefore, there is provided an antigen-binding protein, optionally an isolated antigen-binding protein, that is capable of binding to SARS-CoV-2 including portions thereof.
In various embodiments, the antigen-binding protein is capable of binding specifically to SARS-CoV-2. In various embodiments, the antigen-binding protein is specific to SARS-CoV-2. In various examples, the antigen-binding protein is capable of binding to SARS-CoV-2 or portions thereof in a dose-dependent manner. In various embodiments, the antigen-binding protein does not
substantially bind to or cross-react with a virus of another family that is not the coronavirus family.
In various examples, the antigen-binding protein is capable of binding to pseudotyped SARS-CoV-2. In various examples, the antigen-binding protein is capable of binding to live SARS-CoV-2. A “live” SARS-CoV-2 may comprise a wild-type envelope protein. In contrast, a “pseudotyped” virus may comprise a recombinant virus having a different envelope protein from the wild-type envelope protein. In some examples, the spike protein of a pseudotyped virus may contain only 1254 amino acid residues (instead of 1273 amino acid residues) without the last 19 amino acid residues of the C-terminal region. In various examples, the antigen-binding protein is capable of binding to live or pseudotyped SARS-CoV- 2 in solution and/or on surface.
In various embodiments, the antigen-binding protein is capable of recognising/interacting with/binding to portions of SARS-CoV-2 that mediate virus binding to or entry into a potential host cell. In various embodiments, the antigen binding protein is capable of recognising/interacting with/binding to an envelope protein of SARS-CoV-2 or portions thereof. In various embodiments, the antigen binding protein is capable of recognising/interacting with/binding to the spike protein (or spike glycoprotein) of SARS-CoV-2, e.g., a spike protein of SEQ ID NO: 353, or portions thereof. In various embodiments, the antigen-binding protein is capable of recognising/interacting with/binding to the amino-terminal region or an amino-terminal unit of SARS-CoV-2 spike protein or portions thereof. In various embodiments, the antigen-binding protein is capable of recognising/interacting with/binding to the amino-terminal S1 subunit of SARS- CoV-2 spike protein or portions thereof. In various embodiments, the antigen binding protein is capable of recognising/interacting with/binding to the receptor binding domain (RBD) of SARS-CoV-2 spike protein, e.g., a RBD of SEQ ID NO: 354, or portions thereof. In various embodiments, the antigen-binding protein is capable of recognising/interacting with/binding to an epitope or an antigen on SARS-CoV-2.
In various embodiments, the antigen-binding protein has an ECso value (e.g., an average ECso value) of no more than about 100 nM, no more than about
90 nM, no more than about 80 nM, no more than about 70 nM, no more than about 60 nM, no more than about 50 nM, no more than about 40 nM, no more than about 30 nM, no more than about 20 nM, no more than about 19 nM, no more than about 18 nM, no more than about 17 nM, no more than about 16 nM, no more than about 15 nM, no more than about 14 nM, no more than about 13 nM, no more than about 12 nM, no more than about 11 nM, no more than about 10 nM, no more than about 9.5 nM, no more than about 9 nM, no more than about 8.5 nM, no more than about 8 nM, no more than about 7.5 nM, no more than about 7 nM, no more than about 6.5 nM, no more than about 6 nM, no more than about 5.5 nM, no more than about 5 nM, no more than about 4.5 nM, no more than about 4 nM, no more than about 3.5 nM, no more than about 3 nM, no more than about 2.5 nM, no more than about 2 nM, no more than about 1.5 nM, no more than about 1 nM, no more than about 0.95 nM, no more than about 0.9 nM, no more than about 0.85 nM, no more than about 0.8 nM, no more than about 0.75 nM, no more than about 0.7 nM, no more than about 0.65 nM, no more than about 0.6 nM, no more than about 0.55 nM, no more than about 0.5 nM, no more than about 0.45 nM, no more than about 0.4 nM, no more than about 0.35 nM, no more than about 0.3 nM, no more than about 0.25 nM, no more than about 0.2 nM, no more than about 0.15 nM, no more than about 0.1 nM, no more than about 0.095 nM, no more than about 0.09 nM, no more than about 0.085 nM, no more than about 0.08 nM, no more than about 0.075 nM, no more than about 0.07 nM, no more than about 0.065 nM, no more than about 0.06 nM, no more than about 0.055 nM or no more than about 0.05 nM for SARS-CoV-2 e.g., a RBD of SARS-CoV-2. In various embodiments, the antigen-binding protein has an EC50 value (e.g., an average EC50 value) of no more than about 80 nM, optionally no more than about 12 nM, further optionally no more than about 6 nM, further optionally no more than about 3 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 0.1 nM for SARS-CoV-2 e.g., a RBD of SARS-CoV-2. In various embodiments, the antigen-binding protein has an EC50 value (e.g., an average EC50 value) of from about 0.05 nM to about 80 nM, from about 0.05 nM to about 12 nM, from about 0.05 nM to about 10 nM, from about 0.05 nM to about 5 nM,
from about 0.05 nM to about 2 nM, from about 0.05 nM to about 1 nM, from about 0.05 nM to about 0.5 nM, from about 0.05 nM to about 0.2 nM or from about 0.05 nM to about 0.1 nM for SARS-CoV-2 e.g., a RBD of SARS-CoV-2.
In various embodiments, the antigen-binding protein is capable of blocking/interfering/inhibiting/hindering/disrupting one or more of the following function(s) of SARS-CoV-2 spike protein: (i) host cell attachment; (ii) receptor binding; and (iii) mediating host cell membrane and viral membrane fusion.
In various embodiments, the antigen-binding protein is capable of blocking/interfering/inhibiting/hindering/disrupting the attachment of SARS-CoV- 2 to a host cell.
In various embodiments, the antigen-binding protein is capable of blocking/interfering/inhibiting/hindering/disrupting the binding of SARS-CoV-2 to a cell entry receptor. In various embodiments, the cell entry receptor comprises angiotensin converting enzyme II (ACE2). In various embodiments, the cell entry receptor comprises human ACE2.
In various embodiments, the antigen-binding protein has a potent blocking/interfering/inhibiting/hindering/disrupting action against the binding of SARS-CoV-2 to the cell entry receptor. In various embodiments, the associated EC50 value of the antigen-binding protein is from about 0.1 nM to about 360 nM, from about 0.3 nM to about 100 nM, from about 0.3 nM to about 50 nM, from about 0.3 nM to about 25 nM, from about 0.3nM to about 2.5 nM, from about 0.3 nM to about 2.0nM or from about 0.3 nM to about 1 .5 nM. In some embodiments, the associated EC50 value of the antigen-binding protein is no more than about 2.5 nM, no more than about 2.0 nM, or no more than about 1 .5 nM.
In various embodiments, the antigen-binding protein is capable of neutralising or mediating (or initiating) the neutralisation of SARS-CoV-2. The neutralizing antigen-binding protein may substantially prevent, block, inhibit or hinder the binding or association of SARS-CoV-2 to a cell (e.g., the binding or association of a RBD of a SARS-CoV-2 to a cell entry receptor ACE2 of a cell), thereby interrupting a biological response that would otherwise result from the interaction of SARS-CoV-2 with the cell. The neutralizing antigen-binding protein may substantially prevent, block, inhibit or hinder SARS-CoV-2 from infecting a
cell. In various examples, the antigen-binding protein is capable of substantially neutralising or mediating (or initiating) the neutralisation of SARS-CoV-2 in a dose-dependent manner. In various embodiments, the antigen-binding protein is capable of substantially neutralising or mediating the neutralisation of SARS- CoV-2 entry into cells, optionally human cells, further optionally ACE2 expressing/presenting cells. In various embodiments therefore, the antigen binding protein comprises a SARS-CoV-2 neutralising antigen binding protein.
In various embodiments, the antigen-binding protein has a potent neutralising action against cell entry of SARS-CoV-2. In various embodiments, the associated ECso value of the antigen-binding protein is from about 5 ng/ml to about 17000 ng/ml, from about 6 ng/ml to about 1000 ng/ml, from about 6 ng/ml to about 100 ng/ml, from about 6 ng/ml to about 60 ng/ml, from about 6 ng/ml to about 30 ng/ml or from about 6 ng/ml to about 12 ng/ml. In some embodiments, the associated ECso value of the antigen-binding protein is no more than about 60 ng/ml, no more than about 30 ng/ml, or no more than about 12 ng/ml.
In various embodiments, the antigen-binding protein has an ICso value (e.g., an average ICso value) of no more than about 360 nM, no more than about 350 nM, no more than about 300 nM, no more than about 200 nM, no more than about 100 nM, no more than about 90 nM, no more than about 80 nM, no more than about 70 nM, no more than about 60 nM, no more than about 50 nM, no more than about 40 nM, no more than about 30 nM, no more than about 20 nM, no more than about 10 nM, no more than about 9.5 nM, no more than about 9 nM, no more than about 8.5 nM, no more than about 8 nM, no more than about 7.5 nM, no more than about 7 nM, no more than about 6.5 nM, no more than about 6 nM, no more than about 5.5 nM, no more than about 5 nM, no more than about 4.5 nM, no more than about 4 nM, no more than about 3.5 nM, no more than about 3 nM, no more than about 2.5 nM, no more than about 2 nM, no more than about 1 .5 nM, no more than about 1 nM, no more than about 0.95 nM, no more than about 0.9 nM, no more than about 0.85 nM, no more than about 0.8 nM, no more than about 0.75 nM, no more than about 0.7 nM, no more than about 0.65 nM, no more than about 0.6 nM, no more than about 0.55 nM, no more than about 0.5 nM, no more than about 0.45 nM, no more than
about 0.4 nM, no more than about 0.35 nM, no more than about 0.3 nM, no more than about 0.25 nM, no more than about 0.2 nM, no more than about 0.15 nM, no more than about 0.1 nM or no more than about 0.05 nM for an inhibition of binding between SARS-CoV-2 and a cell, optionally for an inhibition of binding between a spike protein of SARS-CoV-2 and an ACE2 of a cell. In various embodiments, the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of no more than about 360 nM, optionally no more than about 180 nM, further optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 5 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM or further optionally no more than about 0.5 nM for an inhibition of binding between SARS-CoV-2 and a cell, optionally for an inhibition of binding between a spike protein of SARS-CoV-2 and an ACE2 of a cell. In various embodiments, the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of from about 0.05 nM to about 360 nM, from about 0.05 nM to about 200 nM, from about 0.1 nM to about 100 nM, from about 0.1 nM to about 50 nM, from about 0.1 nM to about 10 nM, from about 0.1 nM to about 3 nM, from about 0.1 nM to about 1 .2 nM or from about 0.1 nM to about 0.5 nM for an inhibition of binding between SARS-CoV-2 and a cell, optionally for an inhibition of binding between a spike protein of SARS-CoV-2 and an ACE2 of a cell.
In various embodiments, the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of no more than about 20,000 ng/ml, no more than about 10,000 ng/ml, no more than about 5000 ng/ml, no more than about 4000 ng/ml, no more than about 3000 ng/ml, no more than about 2000 ng/ml, no more than about 1000 ng/ml, no more than about 900 ng/ml, no more than about 800 ng/ml, no more than about 700 ng/ml, no more than about 600 ng/ml, no more than about 500 ng/ml, no more than about 450 ng/ml, no more than about 400 ng/ml, no more than about 350 ng/ml, no more than about 300 ng/ml, no more than about 250 ng/ml, no more than about 200 ng/ml, no more than about 150 ng/ml, no more than about 100 ng/ml, no more than about 95 ng/ml, no more than about 90 ng/ml, no more than about 85 ng/ml, no more than about 80 ng/ml, no more
than about 75 ng/ml, no more than about 70 ng/ml, no more than about 65 ng/ml, no more than about 60 ng/ml, no more than about 55 ng/ml, no more than about 50 ng/ml, no more than about 45 ng/ml, no more than about 40 ng/ml, no more than about 35 ng/ml, no more than about 30 ng/ml, no more than about 25 ng/ml, no more than about 20 ng/ml, no more than about 15 ng/ml, no more than about 10 ng/ml or no more than about 5 ng/ml for an inhibition of SARS-CoV-2 entry into a cell, e.g., pseudotyped SARS-CoV-2 entry into an ACE2-expressing cell. In various embodiments, the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of no more than about 20,000 ng/ml, optionally no more than about 10,000 ng/ml, further optionally no more than about 5000 ng/ml, further optionally no more than about 2500 ng/ml, further optionally no more than about 1000 ng/ml, further optionally no more than about 500 ng/ml, further optionally no more than about 250 ng/ml, further optionally no more than about 100 ng/ml, further optionally no more than about 50 ng/ml, further optionally no more than about 25 ng/ml or further optionally no more than about 10 ng/ml for an inhibition of SARS-CoV-2 entry into a cell, e.g., pseudotyped SARS-CoV-2 entry into an ACE2-expressing cell. In various embodiments, the antigen-binding protein has an IC50 value (e.g., an average IC50 value) of from about 5 ng/ml to about 20,000 ng/ml, from about 5 ng/ml to about 10,000 ng/ml, from about 5 ng/ml to about 1000 ng/ml, from about 5 ng/ml to about 500 ng/ml, from about 5 ng/ml to about 100 ng/ml, from about 5 ng/ml to about 50 ng/ml, from about 5 ng/ml to about 25 ng/ml or from about 5 ng/ml to about 10 ng/ml for an inhibition of SARS-CoV-2 entry into a cell, e.g., pseudotyped SARS-CoV-2 entry into an ACE2-expressing cell.
In various embodiments, the antigen-binding protein has a KD value (e.g., an average KD value) of no more than about 150 nM, no more than about 100 nM, no more than about 50 nM, no more than about 10 nM, no more than about 9 nM, no more than about 8 nM, no more than about 7 nM, no more than about
6 nM, no more than about 5 nM, no more than about 4 nM, no more than about
3 nM, no more than about 2 nM, no more than about 1 nM, no more than about
0.5 nM, no more than about 0.45 nM, no more than about 0.4 nM, no more than about 0.35 nM, no more than about 0.3 nM, no more than about 0.25 nM, no more
than about 0.2 nM, no more than about 0.15 nM, no more than about 0.1 nM, no more than about 50 pM, no more than about 10 pM, no more than about 5 pM or no more than about 1 pM for SARS-CoV-2, e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry. In various embodiments, the antigen binding protein has a KD value (e.g., an average KD value) of no more than about 130 nM, optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 1 pM for SARS-CoV-2, e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry. In various embodiments, the antigen-binding protein has a KD value (e.g., an average KD value) of less than about 1 x 10 12 M for SARS-CoV-2, e.g., a RBD of SARS-CoV-2 e.g., as measured by Bio-Layer Interferometry.
In various embodiments, the antigen-binding protein is capable of inhibiting virus-cell fusion and/or cell-cell fusion mediated by SARS-CoV-2.
In some embodiments, the antigen-binding protein does not bind to, interact with or cross-react with other human coronaviruses (other than SARS-CoV-2). In some embodiments, the antigen-binding protein does not cross-react with severe acute respiratory syndrome coronavirus (SARS-CoV). Advantageously, embodiments of the antigen-binding protein have high specificity for SARS-CoV- 2.
In some embodiments, the antigen-binding protein is capable of binding to or interacting with other human coronaviruses (other than SARS-CoV-2). In some embodiments, the antigen-binding protein is capable of binding to or interacting with SARS-CoV in addition to SARS-CoV-2. Thus, embodiments of the antigen binding protein are capable of binding to an epitope that is conserved between SARS-CoV and SARS-CoV-2. Embodiments of the antigen-binding protein may be useful for detecting, binding to or neutralising two or more human coronaviruses such as SARS-CoV and SARS-CoV-2. In some embodiments, the antigen-binding protein binds to SARS-CoV-2 more strongly than SARS-CoV. In some embodiments, the antigen-binding protein has a higher affinity for SARS-
CoV-2 than SARS-CoV. In some embodiments, the antigen-binding protein neutralises SARS-CoV-2 by a greater extent as compared to SARS-CoV.
In various embodiments the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising one or more mutations. In various embodiments, the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising no more than about 50, no more than about 40, no more than about 30, no more than about 20, no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s).
In various embodiments, the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising a mutated spike protein, for example, a mutated spike protein comprising no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s). In various embodiments, the antigen-binding protein is capable of binding to and/or neutralizing SARS-CoV-2 comprising a mutated RBD, for example, a mutated RBD comprising no more than about 10, no more than about nine, no more than about eight, no more than about seven, no more than about six, no more than about five, no more than about four, no more than about three, no more than about two or no more than about one mutation(s).ln various embodiments the antigen-binding protein is capable of binding to and/or neutralizing at least about two, at least about three, at least about four, at least about five, at least about six, at least about seven or more of the following SARS-CoV-2: a SARS-CoV-2 comprising a wild-type spike protein (e.g., a spike protein of SEQ ID NO: 353) or a wild-type RBD (e.g., a RBD of SEQ ID NO: 354), a SARS-CoV-2 comprising a mutated spike protein with no more than about two mutations, a SARS-CoV-2 comprising a mutated spike protein with no more than about one mutation, a SARS-CoV-2 comprising a N439K mutation in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 360), a SARS-CoV-2 comprising a V483A mutation in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 355), a SARS-
CoV-2 comprising a G476S mutation in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 358), a SARS-CoV-2 comprising a S494P mutation in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 359), a SARS-CoV-2 comprising a V483I mutation in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 356), a SARS-CoV-2 comprising L455I and F456V mutations in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 357) and a SARS-CoV- 2 comprising a D614G mutation in the spike protein or RBD (e.g., a spike protein of SEQ ID NO: 361 ). N439K, V483A, G476S, S494P, V483I, L455I/F456V and D614G mutations are all mutations occurring in the spike protein of SARS-CoV- 2, with the first six of them (i.e., N439K, V483A, G476S, S494P, V483I and L455I/F456V) occurring in the RBD and D614G occurring outside the RBD.
In various embodiments, the antigen-binding protein comprises an amino acid sequence corresponding to a sequence in Tables 1 -4. In various embodiments, the antigen-binding protein comprises an amino acid sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8% or at least about 99.9% with a sequence in Tables 1 -4. In some embodiments, the antigen-binding protein comprises an amino acid sequence that differs by about one, about two, about three, about four, about five, about six, about seven, about eight, about nine, about ten or more amino acids with a sequence in Tables 1 -4.
In various embodiments, the antigen-binding protein comprises one or more amino acid region or complementarity determining region corresponding to a CDR identified in Tables 2 and 4. In various embodiments, the antigen-binding protein comprises one or more amino acid region or complementarity determining region sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at
least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence similarity/identity with a CDR or a combination of about two, about three, about four, about five or about six CDRs identified in Tables 2 and 4. In some embodiments, the antigen-binding protein comprises an amino acid region or CDR that differs by about one, about two, about three, about four, about five, about six, about seven, about eight, about nine, about ten or more amino acids with a CDR or a combination of about two, about three, about four, about five or about six CDRs identified in Tables 2 and 4.
In various embodiments, the antigen-binding protein comprises an amino acid region, a light chain variable domain sequence and/or a heavy chain variable domain sequence corresponding to a light chain variable domain sequence and/or a heavy chain variable domain sequence identified in Tables 1 and 3. In various embodiments, the antigen-binding protein comprises an amino acid region or a light chain variable domain and/or a heavy chain variable domain sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8% or at least about 99.9% with a light chain variable domain and/or a heavy chain variable domain identified in Tables 1 and 3. In some embodiments, the antigen-binding protein comprises an amino acid region or a light chain variable domain and/or a heavy chain variable domain that differs by about one, about two, about three, about four, about five, about six, about seven, about eight, about nine, about ten or more amino acids with a light chain variable domain and/or a heavy chain variable domain identified in Tables 1 and 3.
In various embodiments, the antigen-binding protein comprises one of more of the following: a light-chain CDR1 selected from the group consisting of: SEQ ID NO: 28- 50 and conservative sequence variants thereof;
a light-chain CDR2 selected from the group consisting of: SEQ ID NO: 51 - 64 and conservative sequence variants thereof; a light-chain CDR3 selected from the group consisting of: SEQ ID NO: 65- 90 and conservative sequence variants thereof; a heavy-chain CDR 1 selected from the group consisting of: SEQ ID NO: 117-132 and conservative sequence variants thereof; a heavy-chain CDR 2 selected from the group consisting of: SEQ ID NO: 133-148 and conservative sequence variants thereof; and a heavy-chain CDR 3 selected from the group consisting of: SEQ ID NO: 149-173 and conservative sequence variants thereof.
In various embodiments, the antigen-binding protein comprises the following CDR sequences:
1A5 light-chain CDR1 : SGSIASHY (SEQ ID NO: 28) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYGSGFVV (SEQ ID NO: 65) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISGSGGST (SEQ ID NO: 133) heavy-chain CDR3: AKDYFRWL (SEQ ID NO: 149);
1A8 light-chain CDR1 : QSVLYSSNNKNY (SEQ ID NO: 29) light-chain CDR2: WAS (SEQ ID NO: 52) light-chain CDR3: QQYYGTPYT (SEQ ID NO: 66) heavy-chain CDR1 : GVSISSRSDH (SEQ ID NO: 118) heavy-chain CDR2: ISYSGST (SEQ ID NO: 134) heavy-chain CDR3: ARLASLYSTFDI (SEQ ID NO: 150);
1 B11 light-chain CDR1 : QSLLYSNGYNY (SEQ ID NO: 31 ) light-chain CDR2: LGS (SEQ ID NO: 53)
light-chain CDR3: MQALQTPYT (SEQ ID NO: 68) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARDNPGYSSSWSPNWFDP (SEQ ID NO: 152);
1 B2 light-chain CDR1 : GGRIATNY (SEQ ID NO: 30) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSHDTSRQAV (SEQ ID NO: 67) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ATGEMAGDFDY (SEQ ID NO: 151);
1C2 light-chain CDR1 : QSLLHSNGFTY (SEQ ID NO: 32) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQGIQTPLT (SEQ ID NO: 69) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSSAGTFYFDY (SEQ ID NO: 153);
1C3 light-chain CDR1 : SGSIASYY (SEQ ID NO: 33) light-chain CDR2: EDD (SEQ ID NO: 54) light-chain CDR3: QSFDSSIQHVV (SEQ ID NO: 70) heavy-chain CDR1 : GYPFSTYY (SEQ ID NO: 121) heavy-chain CDR2: IDPSGGST (SEQ ID NO: 137) heavy-chain CDR3: ARVAEQHLDYYFDY (SEQ ID NO: 154);
1 D12 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 )
light-chain CDR3: QSYDGSAWV (SEQ ID NO: 71) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARLEWLRGAFDI (SEQ ID NO: 155);
1 E5 light-chain CDR1 : SGRIASNY (SEQ ID NO: 35) light-chain CDR2: EDT (SEQ ID NO: 55) light-chain CDR3: QSFDSGNQRVV (SEQ ID NO: 72) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARALVGKWLLLRGFDY (SEQ ID NO: 156);
1 F4 light-chain CDR1 : SSNIGRNF (SEQ ID NO: 36) light-chain CDR2: RNN (SEQ ID NO: 56) light-chain CDR3: QSYDNSLFVV (SEQ ID NO: 73) heavy-chain CDR1 : GDSVSSNSAA (SEQ ID NO: 122) heavy-chain CDR2: TYYRSKWYN (SEQ ID NO: 139) heavy-chain CDR3: AREKGIEGPAFDP (SEQ ID NO: 157);
1 H7 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDENIRV (SEQ ID NO: 74) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGRWLRGAFDI (SEQ ID NO: 158);
2C10 light-chain CDR1 : SSTIGTNY (SEQ ID NO: 38) light-chain CDR2: DNY (SEQ ID NO: 58)
light-chain CDR3: GTWDSRLSVGV (SEQ ID NO: 76) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INYRGNT (SEQ ID NO: 142) heavy-chain CDR3: ARWDGGNSVDH (SEQ ID NO: 160);
2C12 light-chain CDR1 : QSINIY (SEQ ID NO: 39) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYITPQT (SEQ ID NO: 77) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARGGRFHYYYYAMDV (SEQ ID NO: 161);
2C9 light-chain CDR1 : QTVGGSY (SEQ ID NO: 37) light-chain CDR2: GAS (SEQ ID NO: 57) light-chain CDR3: QQYASSRT (SEQ ID NO: 75) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPILGIA (SEQ ID NO: 141) heavy-chain CDR3: ARHGGVGATTGYYYMDV (SEQ ID NO: 159);
2D3 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ASSGWLRGAFDI (SEQ ID NO: 162);
2G7 light-chain CDR1 : GGTIGSNY (SEQ ID NO: 40) light-chain CDR2: EDN (SEQ ID NO: 51 )
light-chain CDR3: QSYDSSNRV (SEQ ID NO: 79) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGKWLRGAFDI (SEQ ID NO: 163);
2H4 light-chain CDR1 : EFIGDD (SEQ ID NO: 41 ) light-chain CDR2: EAR (SEQ ID NO: 60) light-chain CDR3: LQYDSFPLT (SEQ ID NO: 80) heavy-chain CDR1 : GYTFSGYY (SEQ ID NO: 124) heavy-chain CDR2: VNPNSGGT (SEQ ID NO: 144) heavy-chain CDR3: ARGGRPGL P A AG Y I D Y (SEQ ID NO: 164);
3A11 light-chain CDR1 : SSDVGTYNY (SEQ ID NO: 42) light-chain CDR2: DVS (SEQ ID NO: 61 ) light-chain CDR3: ASFSSSSTLLV (SEQ ID NO: 81 ) heavy-chain CDR1 : GYTFSTYD (SEQ ID NO: 125) heavy-chain CDR2: ISTYNGDT (SEQ ID NO: 145) heavy-chain CDR3: ARGDSSVGYEYFQH (SEQ ID NO: 165);
3C5 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGSLSGYY (SEQ ID NO: 126) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 )
light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3D2 light-chain CDR1 : SGSIARNY (SEQ ID NO: 43) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDRTNKRV (SEQ ID NO: 82) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3E9 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDSSNLQWV (SEQ ID NO: 84) heavy-chain CDR1 : GYTFTSYG (SEQ ID NO: 127) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARGIITMISDY (SEQ ID NO: 167);
3F1 light-chain CDR1 : GGSIADNF (SEQ ID NO: 45) light-chain CDR2: DYS (SEQ ID NO: 62) light-chain CDR3: QSYDISNPV (SEQ ID NO: 85) heavy-chain CDR1 : GFTFSSYG (SEQ ID NO: 128) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARDGGGGMDV (SEQ ID NO: 168);
3F11 light-chain CDR1 : HSDGRNY (SEQ ID NO: 46) light-chain CDR2: DTS (SEQ ID NO: 63)
light-chain CDR3: HQFRRSVST (SEQ ID NO: 86) heavy-chain CDR1 : GYTFSSSG (SEQ ID NO: 129) heavy-chain CDR2: ISTYNGNT (SEQ ID NO: 146) heavy-chain CDR3: ATSIAVAGIDY (SEQ ID NO: 169);
3H11 light-chain CDR1 : SGNIASYY (SEQ ID NO: 48) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDRPNHVV (SEQ ID NO: 88) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSVVPAAIYDYYYGMDV (SEQ ID NO: 171);
3H7 light-chain CDR1 : TDSIASNY (SEQ ID NO: 47) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDGVNRGVI (SEQ ID NO: 87) heavy-chain CDR1 : GFTFSNYG (SEQ ID NO: 130) heavy-chain CDR2: IWHDGTNK (SEQ ID NO: 147) heavy-chain CDR3: VRDQGAGVWNGYSY (SEQ ID NO: 170);
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172);
6F8 light-chain CDR1 : QTINNNY (SEQ ID NO: 50) light-chain CDR2: DAS (SEQ ID NO: 64)
light-chain CDR3: QQYGSSPRT (SEQ ID NO: 90) heavy-chain CDR1 : GFTVSSNY (SEQ ID NO: 132) heavy-chain CDR2: IYSGGST (SEQ ID NO: 148) heavy-chain CDR3: ARDYGDYYFDY (SEQ ID NO: 173); or conservative sequence variants thereof (e.g., an antigen-binding protein comprising one CDR sequence that is a conservative sequence variant of one CDR sequence of 1A5 and five CDR sequences that are identical to the other five CDR sequences of 1A5, an antigen-binding protein comprising two CDR sequences that are conservative sequence variants of two CDR sequences of 1 B2 and three CDR sequences that are identical to the other three CDR sequences of 1 B2 etc.).
In various embodiments, antigen-binding protein comprises a light-chain variable region comprising any one of SEQ ID NO: 1 -27 or a sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence similarity/identity thereto; and/or a heavy-chain variable region comprising any one of SEQ ID NO: 91 -116 or a sequence sharing at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence similarity/identity thereto.
In various embodiments, there is provided a combination of about two, about three, about four, about five, about six, about seven, about eight, about nine, about 10, about 11 , about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21 , about 22, about 23, about 24, about 25, about 26 or about 27 antigen-binding proteins. In various embodiments, the antigen-binding proteins in the combination comprises the CDR sequences of one or more of the following antigen-binding proteins: 1 A5, 1 A8, 1 B2, 1 B11 , 1 C2, 1 C3, 1 D12, 1 E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6 and 6F8. In various embodiments, the antigen-binding proteins in the combination is selected from the group consisting of: 1A5, 1A8, 1 B2, 1 B11 , 1 C2, 1C3, 1 D12, 1 E5, 1 F4, 1 H7, 2C9, 2C10, 2C12,
2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, 6F8 and combinations thereof.
In various embodiments, there is provided a combination of two antigen binding proteins, wherein a first antigen-binding protein comprises the following CDR sequences:
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172) and a second antigen-binding protein comprises the following CDR sequences:
3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
In various embodiments, there is provided a composition comprising the antigen-binding protein or the combination. In various embodiments, the composition comprises a therapeutically effective amount of the antigen-binding protein or the combination e.g., for use in therapy. In various embodiments, the composition further comprises a suitable carrier, adjuvant, diluent and/or excipient. In various embodiments, the composition comprises a therapeutic composition.
In various embodiments, the composition comprises a pharmaceutical composition. The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier and/or pharmaceutically active compound. In one embodiment, there is provided a pharmaceutical composition comprising the antigen-binding protein or the combination, and a pharmaceutically acceptable carrier.
In various embodiments, there is provided the antigen-binding protein or the combination for use in therapy. In various embodiments, there is provided the antigen-binding protein or the combination when used in therapy.
In various embodiments, there is provided the antigen-binding protein or the combination as a detecting agent, e.g., for detecting SARS-CoV-2. In various embodiments, there is provided the antigen-binding protein or the combination as a diagnostic agent e.g., for SARS-CoV-2.
In various embodiments, there is provided the antigen-binding protein or the combination for use in therapy, or as a diagnostic agent.
In various embodiments, there is provided a polynucleotide/nucleotide sequence encoding for the antigen-binding protein. The polynucleotide/nucleotide sequence may comprise DNA, cDNA and/or RNA. In various embodiments, the polynucleotide/nucleotide sequence comprises one or more sequences set forth in Tables 5-8 or a sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence identity thereto. In various embodiments, the polynucleotide/nucleotide sequence comprises one of more of SEQ ID NO: 174-351 or a sequence sharing at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 85%, at least about 95% or at least about 99% sequence identity thereto.
In various embodiments, there is provided a vector comprising the polynucleotide/nucleotide sequence. In various embodiments, the vector is selected from the group consisting of a phage, a plasmid, a viral particle, a baculovirus, a yeast plasmid, a lipid based vehicle, a polymer microsphere, a liposome, and a cell based vehicle, a colloidal gold particle, lipopolysaccharide, polypeptide, polysaccharide, a viral vehicle, an adenovirus, a retrovirus, a
lentivirus, an adeno-associated viruses, a herpesvirus, a vaccinia virus, a foamy virus, a cytomegalovirus, a Semliki forest virus, a poxvirus, a pseudorabies virus, an RNA virus vector, a DNA virus vector and a vector derived from a combination of a plasmid and a phage DNA. In various embodiments, the polynucleotide/nucleotide sequence is operatively linked to an expression control sequence(s) to direct peptide synthesis. In various embodiments, the vector comprises one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells. In various embodiments, there is provided a host cell transfected with or comprising the vector. The host cell may comprise a bacterial cell, a yeast cell, an animal cell e.g., a mammalian cell and/or a plant cell.
In various embodiments, there is provided a bacteriophage, a host cell or a non-human organism comprising the polynucleotide/nucleotide sequence. In some embodiments, the non-human organism comprises a mammal. In some embodiments, the non-human organism comprises a murine.
In various embodiments, there is provided a method of treating SARS-CoV- 2 infection in a subject, the method comprising administering the antigen-binding protein, the combination or the composition (e.g., pharmaceutical composition) to the subject. In various embodiments, where the SARS-CoV-2 comprises a V483A mutation, the method comprises administering the subject a combination of two antigen-binding proteins, wherein a first antigen-binding protein comprises the following CDR sequences:
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172)
and a second antigen-binding protein comprises the following CDR sequences:
3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
The method may be performed in vitro or ex vivo or in vivo.
In various embodiments, there is provided use of the antigen-binding protein, the combination or the composition (e.g., pharmaceutical composition) in the manufacture of a medicament for the treatment of SARS-CoV-2 infection in a subject.
In various embodiments, there is provided a method of detecting the presence of SARS-CoV-2 in a sample, the method comprising: contacting the sample with the agent, e.g., the antigen-binding protein and detecting a binding event involving the agent, e.g., the antigen-binding protein. In various embodiments, the detection of a binding event is indicative of the presence of SARS-CoV-2 in the sample. In various embodiments, no detection of any binding events involving the agent, e.g., the antigen-binding protein is indicative of the absence of SARS-CoV-2 in the sample. In various embodiments, the antigen binding protein may be labelled e.g., coupled to a label. In one embodiment, the antigen-binding protein is labelled with horseradish peroxidase (FIRP) and tetramethylbenzidine (TMB) substrate may be added for visualization. It will be appreciated that other suitable labels and substrates may also be used for detecting a binding event. In various embodiments, there is provided the antigen binding protein when used in detecting the presence of SARS-CoV-2 in a sample.
The sample may be obtained from any source. For example, the sample may be a biological sample, a pharmaceutical sample, an environmental sample,
a food sample etc. In some embodiments, the sample comprises a biological sample. Examples of biological samples include, but are not limited to blood, serum, plasma, sputum, saliva, lavage fluid (e.g. bronchial lavage fluid, alveolar lavage fluid and bronchoalveolar lavage fluid), sputum, nasal fluid/swab/wash/aspirate, anterior nares fluid/swab/wash/aspirate, nasal mid turbinate fluid/swab/wash/aspirate, pharyngeal fluid/swab/wash/aspirate, nasopharyngeal fluid/swab/wash/aspirate, oropharyngeal fluid/swab/wash/aspirate, tissue biopsy e.g. lung biopsy, cerebrospinal fluid, urine, faeces, stool, anal swab, semen, sweat, tears, processed fractions thereof and the like.
In various embodiments, there is provided a method of identifying an agent, optionally an antigen-binding protein, that is capable of recognising/interacting with/binding to SARS-CoV-2.
In some embodiments, the method comprises contacting a candidate compound with a SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/a SARS-CoV-2 spike protein RBD domain, wherein where the candidate compound binds to the SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain, the candidate agent is identified as a potential agent. In some embodiments, the method comprises use of phage display technology to identify the agent. In some embodiments, the method comprises an enzyme-linked immunosorbent assay (ELISA) method. In various embodiments, the ELISA method comprises immobilizing SARS-CoV-2 spike protein RBD domain or antigen to a surface, adding a candidate compound to the surface under conditions suitable for binding, and determining a binding between the SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS- CoV-2 spike protein RBD domain or antigen and the candidate compound. In some embodiments, the SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain or antigen is tagged (e.g., with biotin). In some embodiments, the method comprises determining whether a candidate compound is capable of blocking a binding between SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD
domain and ACE2 protein, wherein where the candidate compound is capable of blocking a binding between SARS-CoV-2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain and ACE2 protein, the candidate compound is identified as a potential agent. In some embodiments, the method comprises providing a candidate compound that is bound to SARS-CoV- 2/a SARS-CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain, and contacting the bound candidate compound with ACE2 protein, wherein where the bound candidate compound and/or the SARS-CoV-2/a SARS- CoV-2 spike protein (or glycoprotein)/ SARS-CoV-2 spike protein RBD domain does not bind to the ACE2 protein, the candidate compound is identified as a potential agent.
In some embodiments, the method comprises determining whether a candidate compound is capable of neutralising cell entry of SARS-CoV-2 or potions thereof, wherein where the candidate compound is capable of neutralising cell entry of SARS-CoV-2 or portions thereof, the candidate compound is identified as a potential agent. In some embodiments, the method comprises contacting SARS-CoV-2 (or a pseudotyped SARS-CoV-2) or potions thereof with ACE-expressing cells in the presence of the candidate compound, and determining an amount of entry of the SARS-CoV-2 or portions thereof into the ACE-expressing cells, wherein where the amount of entry of the SARS-CoV- 2 or portions thereof into the ACE-expressing cells is substantially low/reduced, the candidate compound is identified as a potential antigen-binding protein. In various embodiments, the determining step comprises performing a luciferase assay. In some embodiments, the candidate compound is pre-incubated with the
SARS-CoV-2 or potions thereof before contact with the ACE-expressing cells.
In various embodiments, the method further comprises generating a library of sequences, optionally Fab sequences, to obtain the candidate compounds. In some embodiments, the library comprises a phase display library. In various embodiments, there is provided a product or a method as described herein.
Table 1. Light chain variable domain amino acid sequences for human lgG1
Table 2. Light-chain CDR amino acid sequences for human lgG1
Table 3. Heavy chain variable domain amino acid sequences for human lgG1
Table 4. Heavy-chain CDR amino acid sequences for human lgG1
Table 5. Light chain variable domain nucleotide sequences for human lgG1
Table 6. Light-chain CDR nucleotide sequences for human lgG1
Table 7. Heavy chain variable domain nucleotide sequences for human lgG1
Table 8. Heavy-chain CDR nucleotide sequences for human lgG1
It will be appreciated by a person skilled in the art that other variations and/or modifications may be made to the embodiments disclosed herein without departing from the spirit or scope of the disclosure as broadly described. For example, in the description herein, features of different exemplary embodiments may be mixed, combined, interchanged, incorporated, adopted, modified, included etc. or the like across different exemplary embodiments. The present embodiments are, therefore, to be considered in all respects to be illustrative and not restrictive.
DETAILED DESCRIPTION OF FIGURES
Example embodiments of the disclosure will be better understood and readily apparent to one of ordinary skill in the art from the following discussions and if applicable, in conjunction with the figures. It should be appreciated that other modifications within the purview of the skilled person in the art may be made
without deviating from the scope of the invention. Example embodiments are not necessarily mutually exclusive as some may be combined with one or more embodiments to form new exemplary embodiments. The example embodiments should not be construed as limiting the scope of the disclosure. FIG. 1 shows binding ELISA of supernatants of 27 Fab clones to the antigen protein of biotinylated RBD-mFc. The 27 Fab clones were tested in a binding ELISA assay to assess their antigen binding to biotinylated RBD with a mouse Fc tag and detected by goat-anti-human Fab-HRP. In this assay, the clones were tested with 70 mI of 0.2 pg/ml Biotinylated SARS-CoV-2 RBD_mFC, total 14 ng per well.
FIG. 2 shows blocking of spike RBD protein binding to ACE2-His by supernatants of 27 Fab clones. The 27 Fab clones were tested in an ELISA assay to assess their potency in blocking spike protein binding to the recombinant ACE2 protein (ACE2 with His tag), with an irrelevant Fab clones used as a negative control. In this assay, the clones were tested with 100 mI of 0.5 nM Biotinylated SARS-CoV-2 RBDjTiFC, total 0.05 pmol per well.
FIG. 3 shows binding ELISA of 19 IgGs to the antigen protein of biotinylated RBD-mFc. The 19 antibody clones were tested in a binding ELISA assay to assess their antigen binding to biotinylated RBD with a mouse Fc tag, and detected by anti-human Fc-HRP, with an irrelevant lgG1 used as a negative control antibody. In this assay, the clones were tested with 70 mI of 0.2 pg/ml Biotinylated SARS-CoV-2 RBDJTIFC, total 14 ng per well.
FIG. 4 shows blocking of spike RBD protein binding to ACE2-Fc by 19 IgGs. The 19 antibody clones were tested in an ELISA assay to assess their potency in blocking spike protein binding to the recombinant ACE2 protein (ACE2 with human Fc tag), with an irrelevant IgG used as a negative control antibody. In this assay, the clones were tested with 100 mI of 0.5 nM Biotinylated SARS- CoV-2 RBDJTIFC, total 0.05 pmol per well.
FIG. 5 shows competition of RBD-binding between ACE2 and six IgGs by Bio-Layer Interferometry assay. A weak blocking IgG clone 1C3 (black line) was included as a negative control.
FIG. 6 shows binding avidity of six IgGs to the RBD by Bio-Layer Interferometry assay. A range of IgG concentration from 12.5 nM to 0.39 nM (in 2-fold dilutions) are shown for each IgG. The sensorgrams are in grey lines and curve fittings are in black. Results shown are a representative of two independent experiments.
FIG. 7 shows binding avidity of six IgG antibodies to SARS-CoV-2 (circle) and SARS-CoV (triangle) spike RBD proteins tested by ELISA. The 6 antibody IgG clones 1 F4, 2H4, 3D11 , 3F11 , 5A6 and 6F8 were tested in a binding ELISA assay to assess their antigen binding to biotinylated SARS-CoV-2 RBD with a mouse Fc tag as well as biotinylated SARS-CoV RBD with a His tag and detected by anti-human Fc-HRP. In this assay, the antibodies were tested with 70 mI of 0.2 pg/ml Biotinylated SARS-CoV-2 RBDJTIFC, total 14 ng per well or 70 mI of 0.2 pg/ml Biotinylated SARS-CoV RBDJHis, total 14 ng per well.
FIG. 8 shows epitope binding of 5A6 by Bio-Layer Interferometry (BLI) analysis. Buffer alone, an irrelevant antibody (AB) and 5A6 IgG were included as controls.
FIG. 9 shows binding affinity of five Fab clones to SARS-CoV-2 Spike RBD protein measured by Bio-Layer Interferometry. Fab binding to immobilized Fc- RBD was tested using a range of Fab concentrations from 100 nM to 3.125 nM (in 2-fold dilution). A representative set of measurements is shown with sensorgrams in grey and curve fittings in black.
FIG. 10 shows binding of the 6 IgG antibodies (solid lines) and 5A6 Fab (dashed line,) to the purified SARS-CoV-2 pseudovirus. In this assay, the antibodies were tested with 70 mI of 1 pg/ml of pseudoviral particles, total 70 ng per well.
FIG. 11 shows SARS-CoV-2 pseudotyped virus neutralization assay by 19 IgGs. The 19 antibody clones were tested in a pseudotyped virus neutralization assay to assess their potency in neutralizing pseudotyped virus entry to ACE2 expression cells. FIG. 12 shows neutralization of the SARS-CoV-2 and SARS-CoV pseudotyped viruses by anti-SARS-CoV-2 spike RBD IgG and Fab antibodies. (A) Infection of CHO-ACE2 cells by SARS-CoV-2 pseudovirus were determined
in the presence of 1 F4, 2H4, 3D11 , 3F11, 5A6 and 6F8 IgG antibodies. (B) Infection of CFIO-ACE2 cells by SARS-CoV pseudovirus were determined in the presence of 1 F4, 2H4, 3D11 , 3F11 , 5A6 and 6F8 IgG antibodies. (C) Infection of CFIO-ACE2 cells by SARS-CoV-2 pseudovirus were determined in the presence of 1 F4, 2H4, 3D11 , 3F11 , and 5A6 Fab antibodies. (D) Infection of CHO-ACE2 cells by SARS-CoV pseudovirus were determined in the presence of 1 F4, 2H4, 3D11 , 3F11 , and 5A6 Fab antibodies. Luciferase activities in the CFIO-ACE2 cells were measured, and the percent neutralization was calculated. Data are presented as means ± SEM in triplicates and are representative of two independent experiments. The IC50 values shown in (A) and (C) were calculated by a variable slope four parameter non-linear regression model without or Awith top and bottom constrains set at 100% and 0% respectively.
FIG. 13 shows binding avidity of 5A6 IgG (A) and 3D11 IgG (B) to the wildtype RBD and RBD mutants measured by biolayer interferometry. A range of 5A6 and 3D11 IgG concentrations from 12.5 nM to 0.39 nM (in 2-fold dilutions) are measured for each experiment. The sensorgrams are in grey lines and curve fittings are in black.
FIG. 14 shows neutralization of the SARS-CoV-2 pseudovirus with either wildtype (WT) or RBD mutations by 5A6 (A) and 3D11 (B) IgGs. Data are presented as means ± SEM in triplicates and are representative of two independent experiments.
FIG. 15 shows anti-SARS-CoV-2 Spike RBD IgG antibodies affect trypsin induced cell syncytia formation. Vero E6 cells were transfected with furin recognition mutation of SARS-CoV-2 S-protein (R682RAR to A682AAR)-GFP. After 48 hours, the cell culture medium was changed to DMEM (no serum) and treated with or without antibodies and incubated for 1 hour at 37°C. The cells were then treated with or without trypsin 15 pg/ml for 2 hours at 37°C. Cells were fixed with 4%PFA and stained with DAPI. (A) 5A6 IgG (20 pg/ml), 5A6 Fab (20 pg/ml), 3D11 IgG (20 pg/ml) and 2H4 IgG (10 pg/ml) treated S-protein expressing Vero E6 cell images in 10x, 20x and 40x objective view. Images were taken by Olympus confocal microscope. (B) Dosage response of 5A6 IgG, 5A6 Fab, 3D11 IgG and 2H4 IgG. S-protein expressing Vero E6 were treated with 0, 0.1 , 0.5, 2,
10 g/ml of each antibody. 2x105 cells/sample were used for transfection of 5A6 IgG dosage response. 1.6x105 cells/sample were used for transfection of 5A6 Fab, 2H4 IgG and 3D11 IgG. Data quantification were calculated on syncytia numbers and nuclei number presented in each syncytium. FIG. 16 shows the amino acid sequences of the antibodies as described herein. CDRs are shaded and identified consecutively as CDR1 (first shaded region), CDR2 (second shaded region), and CDR3 (third shaded region).
FIG. 17 shows the nucleotide sequences of the antibodies as described herein. CDRs are shaded and identified consecutively as CDR1 (first shaded region), CDR2 (second shaded region), and CDR3 (third shaded region).
EXPERIMENTAL SECTION
A newly identified novel coronavirus (SARS-CoV-2) is causing pneumonia-associated respiratory syndrome across the world. Epidemiology, genomics, and pathogenesis of the SARS-CoV-2 show high homology with that of SARS-CoV. Current efforts are focusing on development of therapeutic agents for the treatment of coronavirus disease 2019 (COVID-19).
Like SARS-CoV, SARS-CoV-2 also uses the angiotensin converting enzyme II (ACE2) as the cell entry receptor. During infection, the spike glycoprotein on coronavirus envelope is responsible for host cell attachment, receptor binding, and for mediating host cell membrane and viral membrane fusion. The spike protein contains an amino-terminal S1 unit and a carboxyl- terminal S2 subunit. The receptor binding domain (RBD) within the S1 subunit of SARS-CoV-2, which mediates the virus binding to ACE2, is believed to be critical in mediating virus entry into the infected cells. Flence, there is a need to develop therapeutic neutralizing antibodies which can potently bind to RBD of SARS-CoV- 2 spike protein and therefore, to prevent infection by blocking the live viral particles from entering the susceptible cells expressing ACE2. Nucleotide and amino acid sequences for 27 fully human monoclonal antibodies, derivatives thereof, or antigen-binding portions thereof disclosed herein, are able to specifically target/bind SARS-CoV-2 spike protein RBD and
mediate neutralization of virus entry into cells and hence block viral infections. Namely, the antibodies are 1A5, 1A8, 1 B2, 1 B11 , 1C2, 1C3, 1 D12, 1E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, and 6F8. The antibodies, derivatives thereof, or antigen-binding portions thereof disclosed herein are able to specifically target/bind SARS-CoV- 2 spike protein RBD both in solution and on the surface of the pseudotyped virus.
Materials and Methods for converting Fab sequences to IgG antibodies and production of IgG antibodies Fabs were reformatted into human IgG in the pTT5 vector (National
Research Council of Canada) by using In-Fusion FID Cloning kit (Takara Bio) and the IgG antibodies were expressed using ExpiCFIO expression system (Thermo Fisher Scientific) by transient co-transfection of plasmids expressing the heavy and light chain of each antibody clone. Eight days after transfection, ExpiCFIO-S cell suspension was centrifuged for 10 min at 2000 rpm and filtered with 0.22 pm filter to remove the cells and debris. Antibodies were then purified from the culture supernatant using Protein G Agarose (Merck Millipore, Cat#16-266) following the manufacturer’s instructions. After elution, the purified antibodies were dialyzed at 4°C for 4-20 hours against 1x PBS, for 3 times and concentrated to 1-2 mg/ml using 10MWCO Vivaspin 20 (Sartorius, Cat#VS2001).
Antibodies characterisation and tests
Recombinant SARS-CoV-2 spike protein RBD domain with a mouse Fc tag (RBD-mFc) was used to isolate binders from a library of Fab sequences constructed by SlgN, using phage display technology. Out of 570 clones screened, 48 clones showing the positive Fab supernatant binding signals to RBD-mFc and potent antigen-receptor (RBD-ACE2) blocking were sequenced and 27 unique sequences were identified: 1 A5, 1 A8, 1 B2, 1 B11 , 1 C2, 1 C3, 1 D12, 1 E5, 1 F4, 1 H7, 2C9, 2C10, 2C12, 2D3, 2G7, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, and 6F8 (FIG. 16, FIG. 17). The results of binding
ELISA and antigen-receptor blocking ELISA using Fab supernatants for the 27 unique clones were shown in FIG. 1 and FIG. 2, respectively.
Out of 27 unique clones, 19 clones were selected and cloned into lgG1 format for further characterisation: 1A8, 1 B11 , 1C2, 1C3, 1 E5, 1 F4, 2C12, 2H4, 3A11 , 3C5, 3D2, 3D11 , 3E9, 3F1 , 3F11 , 3H7, 3H11 , 5A6, and 6F8.
The 19 antibody clones were tested in ELISA against biotinylated recombinant SARS-CoV-2 spike protein RBD-mFc to assess binding avidity for the target, with an irrelevant lgG1 used as a negative control antibody. In brief, neutravidin was coated at 5pg/ml onto the 96-well ELISA plates in coating buffer overnight at 4°C. After blocking with casein for 2 hours, biotinylated antigen of spike protein RBD-mFc at 0.2 pg/ml was added to the plates and captured by neutravidin during 1-hour incubation at room temperature. After washing with 0.05% PBST for 5 times, 19 antibody IgGs were added at different concentrations with 3 times dilutions and incubated for 1 hour. The wells were then washed again with PBST, followed by addition of HRP conjugated anti-human Fc (1 :3000). The results showed dose dependent antigen binding of all 19 IgG clones (FIG. 3). Next, the 19 antibody clones were tested in an ELISA to assess their abilities in blocking spike protein RBD binding to human ACE2 protein. In brief, the recombinant human ACE2 protein with human Fc tag (ACE2-Fc) was coated onto the 96-well ELISA plates in coating buffer overnight at 4°C and blocked in casein. Then different concentrations of IgGs with 3 times dilutions were pre- incubated with 0.5nM biotinylated spike protein RBD-mFc for 1 hour at room temperature before they were added to the ELISA plates coated with ACE2-Fc. After 1 -hour incubation, the wells were washed with 0.05% PBST for 5 times and HRP conjugated goat-anti-mouse antibody was added at a dilution of 1 :3000. The results in FIG. 4 showed dose dependent blocking of antigen-receptor binding by most lgG1 clones except for 1C3. Out of 18 clones showing capacities of blocking, 5A6 stands out as the most potent clones with a IC50 of 0.3 nM.
Out of the 19 IgGs, 6 antibody clones 1 F4, 2H4, 3D11 , 3F11 , 5A6 and 6F8 also showed potent blocking (FIG. 5) and strong binding avidity (FIG. 6) as measured by the Bio-Layer Interferometry (BLI) assay, and hence were chosen for further detailed characterizations.
To test if any of these six antibody clones cross-react with SARS-CoV RBD, an avidity binding ELISA was performed for both SARS-CoV-2 and SARS-
CoV RBD proteins. Interestingly, only 3D11 was able to cross react with SARS- CoV spike RBD, although the binding was much weaker (FIG. 7). The data suggested that 3D11 recognized a distinct epitope that is different from the other 5 antibodies and is conserved between SARS-CoV-2 and SARS-CoV. Indeed, stepwise binding assay by BLI confirmed that 5A6 and 3D11 had non-overlapping footprints on the RBD while 5A6 shared overlapping epitopes with the other 4 antibodies (FIG. 8).
The present inventors also produced Fab antibodies for clone 1 F4, 2H4, 3D11 , 3F11 and 5A6 and measured their binding affinity to SARS-CoV-2 spike RBD protein by BLI (FIG. 9). A comparison of KD of Fab binding (affinity) and apparent KD of IgG binding (avidity) was also compared and shown in Table 9.
Table 9. KD of Fab and apparent KD of IgG measured by Bio-Layer Interferometry (BLI).
KD of Fab based on 1 :1 Langmuir fitting and apparent KD of IgG based on 1 :2 bivalent analyte fitting of BLI sensorgrams for immobilized Fc-RBD. Values are the Mean ± SD of two independent experiments
Next, the SARS-CoV-2 pseudotyped virus was purified by gradient centrifugation and immobilized the viral particles on the ELISA plates to test if the antibodies as disclosed herein also bind efficiently to the natural RBD displayed on the viral surface. Concentration-dependent binding curves of the 5 IgG clones (1 F4, 2H4, 3D11 , 3F11 , and 5A6) revealed that 5A6 was packed on the viral surface at a much higher density than the other 4 antibodies and 5A6 Fab antibody, as shown by the higher optical signal (FIG. 10).
SARS-CoV-2 pseudotyped virus neutralization assay
The present disclosure tested the function of the 19 IgG antibody clones in neutralizing a pseudotyped virus particles expressing the spike protein of SARS-CoV-2. Briefly, serially diluted IgG antibodies were pre-incubated with an equal volume of pseudotyped virus (12 ng equivalent of p24) for 1 hour before the mixture was added to the monolayer of pre-seeded CHO-ACE2 cells. After 1 hour of pseudotyped virus infection, culture medium was topped up for further incubation of cells for another 48 hours. Then the cells were washed with PBS and lysed in Passive Lysis Buffer and the luciferase assay was performed using the Luciferase Assay System (Promega). The luciferase reading corresponds to the amount of entry of pseudotyped virus particles into the susceptible cells. The results showed that clone 1 F4, 5A6, 3F11 , 2H4 could potently neutralize the cell entry of pseudotyped virus (FIG. 11). Next, a neutralization assay using both SARS-CoV-2 and SARS-CoV pseudotyped virus was performed with the 6 IgG clones 1 F4, 2H4, 3D11 , 3F11 , 5A6 and 6F8. All 6 antibodies showed dose- response neutralization (FIG. 12A). 5A6 strongly neutralized the SARS-CoV-2 pseudovirus with an IC50 of 75.5 ng/ml, followed by 1 F4 (108 ng/ml), 2H4 (109 ng/ml), 6F8 (428.3 ng/ml), and 3F11 (930.4 ng/ml). Interestingly, while 3D11 neutralized SARS-CoV-2 pseudovirus, the neutralization did not follow the typical sigmoidal dose response, suggesting a unique mechanism of inhibition. Consistent with the binding results, none of these antibodies showed significant neutralization to the SARS-CoV pseudotyped virus (FIG. 12B), suggesting high specificity of the antibodies as disclosed herein to the SARS-CoV-2. Interestingly, although 3D11 was able to bind weakly to the SARS-CoV RBD (FIG. 7), it was unable to neutralize the SARS-CoV pseudotyped virus to a significant extent. In addition, neutralization potency of both SARS-CoV-2 and SARS-CoV pseudotyped virus was also performed in the presence of 5 Fab clones 1 F4, 2H4, 3D11 , 3F11 and 5A6 (FIG. 12C and 12D). Sensitivity of 5A6 and 3D11 to RBD mutations
Of more than 40,000 SARS-CoV-2 genomes sequenced, mutations that resulted in amino acid changes in 6 positions at or near the ACE2 interface have
been isolated. These include N439K in 246 samples (244 Scottish, 1 England, 1 Romania), V483A in 30 samples (26 USA/WA, 2 USA/UN, 1 USA/CT, 1 England), G476S in 18 samples (13 USA/WA, 2 USA/OR, 1 USA/ID, 1 USA/CT, 1 Belgium), S494P in 7 samples (3 USA/Ml, 1 England, 1 Spain, 1 India, 1 Sweden), V483I in 2 English samples, and L455I together with F456V in one Brazilian sample.
To determine if any of these mutant viruses could escape neutralization by 5A6 or 3D11 , the recombinant mutant RBD proteins were produced to determine changes in binding avidity. 5A6 IgG had a 4-fold reduction in binding avidity to the V483A mutation while insensitive to the other 5 mutations (FIG. 13A). Consistent with the binding results, 5A6 significantly lost neutralizing capacity against the V483A mutant pseudovirus. However, 5A6 largely retains activity against the other variants, including the D614G mutation that has spread at an alarming rate and become the dominant pandemic strain with a global frequency of 70.5% (GISAID), since its first identification in Europe in March 2020 (FIG. 14A). Interestingly, 3D11 binding was not affected by any of these mutations
(FIG. 13B), but its neutralizing potency against the D614G mutant was partially reduced (FIG. 14B). The difference between the activity profiles of 5A6 and 3D11 could be explained by the different epitopes recognized by each antibody (FIG. 8). Hence, 3D11 could potentially rescue the loss of potency by 5A6 in neutralizing mutant virus strain bearing V483A mutation, when these two antibodies are used as a combination.
5A6 IgG blocks cell-cell fusion
Using a trypsin-triggered cell-cell fusion assay, the impact of the anti- SARS-CoV-2 spike RBD antibodies on syncytia formation was assayed. The 5A6 IgG has an inhibitory effect on syncytia in a dose dependent manner (FIG. 15B), by contrast 2H4 IgG has no significant effect. Surprisingly, 3D11 potentiates cell cell fusion. Furthermore, 5A6 Fabs also enhanced syncytial fusion, albeit weakly (FIG.15A). Thus, it was concluded that 5A6 IgG directly inhibits Spike-mediated fusion, while other receptor blocking antibodies fail to inhibit or even accelerate this process. This observation also suggests that the 5A6 epitope and its binding mode combine efficient receptor blockade and cooperative, avid binding with
conformational trapping of the pre-fusion state, prohibiting the post-fusion transition and preventing both targeted viral fusion and cell-cell fusion mediated by spike protein.
Claims
1. An isolated antigen-binding protein that is capable of binding to and/or neutralizing severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2).
2. The antigen-binding protein according claim 1 , wherein the antigen binding protein is capable of binding to a spike protein of SARS-CoV-2, optionally a receptor-binding domain (RBD) of SARS-CoV-2.
3. The antigen-binding protein according to claim 1 or claim 2, the antigen binding protein comprising one or more of the following properties:
- an EC50 value of no more than about 80 nM, optionally no more than about 12 nM, further optionally no more than about 6 nM, further optionally no more than about 3 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 0.1 nM for SARS-CoV-2;
- an IC50 value of no more than about 360 nM, optionally no more than about 180 nM, further optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 5 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM or further optionally no more than about 0.5 nM for an inhibition of binding between SARS-CoV-2 and a cell;
- an IC50 value of no more than about 20,000 ng/ml, optionally no more than about 10,000 ng/ml, further optionally no more than about 5000 ng/ml, further optionally no more than about 2500 ng/ml, further optionally no more than about 1000 ng/ml, further optionally no more than about 500 ng/ml, further optionally no more than about 250 ng/ml, further optionally no more than about 100 ng/ml, further optionally no more than about 50 ng/ml, further optionally no more than about 25 ng/ml or further optionally no more than about 10 ng/ml for an inhibition of SARS-CoV-2 entry into a cell; and/or
- a KD value of no more than about 130 nM, optionally no more than about 100 nM, further optionally no more than about 50 nM, further optionally no more than about 10 nM, further optionally no more than about 2 nM, further optionally no more than about 1 nM, further optionally no more than about 0.5 nM or further optionally no more than about 1 pM for SARS-CoV-2.
4. The antigen-binding protein according to any one of the preceding claims, wherein the antigen-binding protein does not cross-react with severe acute respiratory syndrome coronavirus (SARS-CoV).
5. The antigen-binding protein according to any one of claims 1-3, wherein the antigen-binding protein is capable of binding to SARS-CoV.
6. The antigen-binding protein according to any one of the preceding claims, the antigen-binding protein comprising a monoclonal antigen-binding protein.
7. The antigen-binding protein according to any one of the preceding claims, the antigen-binding protein comprising a fully human antigen-binding protein.
8. The antigen-binding protein according to any one of claims 1-7, wherein the antigen-binding protein comprises an IgG-like structure.
9. The antigen-binding protein according to any one of claims 1 -7, wherein the antigen-binding protein comprises a Fab fragment.
10. The antigen-binding protein according any one of the preceding claims, wherein the antigen-binding protein is capable of binding to and/or neutralizing about two or more of the following SARS-CoV-2: a SARS- CoV-2 comprising a wild-type spike protein, a SARS-CoV-2 comprising a
mutated spike protein with no more than about two mutations, a SARS- CoV-2 comprising a mutated spike protein with no more than about one mutation, a SARS-CoV-2 comprising a N439K mutation, a SARS-CoV-2 comprising a V483A mutation, a SARS-CoV-2 comprising a G476S mutation, a SARS-CoV-2 comprising a S494P mutation, a SARS-CoV-2 comprising a V483I mutation, a SARS-CoV-2 comprising L455I and F456V mutations and a SARS-CoV-2 comprising a D614G mutation.
11. The antigen-binding protein according any one of the preceding claims, the antigen-binding protein being capable of inhibiting virus-cell fusion and/or cell-cell fusion mediated by SARS-CoV-2.
12. The antigen-binding protein according to any one of the preceding claims, comprising: a light-chain complementarity determining region CDR1 selected from the group consisting of: SEQ ID NO: 28-50 and conservative sequence variants thereof; a light-chain CDR2 selected from the group consisting of: SEQ ID NO: 51-64 and conservative sequence variants thereof; a light-chain CDR3 selected from the group consisting of: SEQ ID NO: 65-90 and conservative sequence variants thereof; a heavy-chain CDR 1 selected from the group consisting of: SEQ ID NO: 117-132 and conservative sequence variants thereof; a heavy-chain CDR 2 selected from the group consisting of: SEQ ID NO: 133-148 and conservative sequence variants thereof; and a heavy-chain CDR 3 selected from the group consisting of: SEQ ID NO: 149-173 and conservative sequence variants thereof.
13. The antigen-binding protein according to any one of the preceding claims, comprising the following CDR sequences:
1A5 light-chain CDR1 : SGSIASHY (SEQ ID NO: 28) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYGSGFVV (SEQ ID NO: 65) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISGSGGST (SEQ ID NO: 133) heavy-chain CDR3: AKDYFRWL (SEQ ID NO: 149);
1A8 light-chain CDR1 : QSVLYSSNNKNY (SEQ ID NO: 29) light-chain CDR2: WAS (SEQ ID NO: 52) light-chain CDR3: QQYYGTPYT (SEQ ID NO: 66) heavy-chain CDR1 : GVSISSRSDH (SEQ ID NO: 118) heavy-chain CDR2: ISYSGST (SEQ ID NO: 134) heavy-chain CDR3: ARLASLYSTFDI (SEQ ID NO: 150);
1 B2 light-chain CDR1 : GGRIATNY (SEQ ID NO: 30) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSHDTSRQAV (SEQ ID NO: 67) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ATGEMAGDFDY (SEQ ID NO: 151);
1 B11 light-chain CDR1 : QSLLYSNGYNY (SEQ ID NO: 31 ) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQALQTPYT (SEQ ID NO: 68)
heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARDNPGYSSSWSPNWFDP (SEQ ID NO: 152);
1 C2 light-chain CDR1 : QSLLHSNGFTY (SEQ ID NO: 32) light-chain CDR2: LGS (SEQ ID NO: 53) light-chain CDR3: MQGIQTPLT (SEQ ID NO: 69) heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSSAGTFYFDY (SEQ ID NO: 153);
1C3 light-chain CDR1 : SGSIASYY (SEQ ID NO: 33) light-chain CDR2: EDD (SEQ ID NO: 54) light-chain CDR3: QSFDSSIQHVV (SEQ ID NO: 70) heavy-chain CDR1 : GYPFSTYY (SEQ ID NO: 121) heavy-chain CDR2: IDPSGGST (SEQ ID NO: 137) heavy-chain CDR3: ARVAEQHLDYYFDY (SEQ ID NO: 154);
1 D12 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDGSAWV (SEQ ID NO: 71 ) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARLEWLRGAFDI (SEQ ID NO: 155);
1 E5 light-chain CDR1 : SGRIASNY (SEQ ID NO: 35) light-chain CDR2: EDT (SEQ ID NO: 55) light-chain CDR3: QSFDSGNQRVV (SEQ ID NO: 72) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ARALVGKWLLLRGFDY (SEQ ID NO: 156);
1 F4 light-chain CDR1 : SSNIGRNF (SEQ ID NO: 36) light-chain CDR2: RNN (SEQ ID NO: 56) light-chain CDR3: QSYDNSLFVV (SEQ ID NO: 73) heavy-chain CDR1 : GDSVSSNSAA (SEQ ID NO: 122) heavy-chain CDR2: TYYRSKWYN (SEQ ID NO: 139) heavy-chain CDR3: AREKGIEGPAFDP (SEQ ID NO: 157);
1 H7 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDENIRV (SEQ ID NO: 74) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGRWLRGAFDI (SEQ ID NO: 158);
2C9 light-chain CDR1 : QTVGGSY (SEQ ID NO: 37) light-chain CDR2: GAS (SEQ ID NO: 57)
light-chain CDR3: QQYASSRT (SEQ ID NO: 75) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPILGIA (SEQ ID NO: 141) heavy-chain CDR3: ARHGGVGATTGYYYMDV (SEQ ID NO: 159);
2C10 light-chain CDR1 : SSTIGTNY (SEQ ID NO: 38) light-chain CDR2: DNY (SEQ ID NO: 58) light-chain CDR3: GTWDSRLSVGV (SEQ ID NO: 76) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INYRGNT (SEQ ID NO: 142) heavy-chain CDR3: ARWDGGNSVDH (SEQ ID NO: 160);
2C12 light-chain CDR1 : QSINIY (SEQ ID NO: 39) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYITPQT (SEQ ID NO: 77) heavy-chain CDR1 : GFTFSSYA (SEQ ID NO: 117) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARGGRFHYYYYAMDV (SEQ ID NO: 161);
2D3 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78) heavy-chain CDR1 : GGTFSSYA (SEQ ID NO: 120) heavy-chain CDR2: IIPIFGTA (SEQ ID NO: 136) heavy-chain CDR3: ASSGWLRGAFDI (SEQ ID NO: 162);
2G7 light-chain CDR1 : GGTIGSNY (SEQ ID NO: 40) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNRV (SEQ ID NO: 79) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARGKWLRGAFDI (SEQ ID NO: 163);
2H4 light-chain CDR1 : EFIGDD (SEQ ID NO: 41 ) light-chain CDR2: EAR (SEQ ID NO: 60) light-chain CDR3: LQYDSFPLT (SEQ ID NO: 80) heavy-chain CDR1 : GYTFSGYY (SEQ ID NO: 124) heavy-chain CDR2: VNPNSGGT (SEQ ID NO: 144) heavy-chain CDR3: ARGGRPGLPAAGYIDY (SEQ ID NO: 164);
3A11 light-chain CDR1 : SSDVGTYNY (SEQ ID NO: 42) light-chain CDR2: DVS (SEQ ID NO: 61 ) light-chain CDR3: ASFSSSSTLLV (SEQ ID NO: 81 ) heavy-chain CDR1 : GYTFSTYD (SEQ ID NO: 125) heavy-chain CDR2: ISTYNGDT (SEQ ID NO: 145) heavy-chain CDR3: ARGDSSVGYEYFQH (SEQ ID NO: 165);
3C5 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNWV (SEQ ID NO: 78)
heavy-chain CDR1 : GGSLSGYY (SEQ ID NO: 126) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3D2 light-chain CDR1 : SGSIARNY (SEQ ID NO: 43) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDRTNKRV (SEQ ID NO: 82) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3D11 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166);
3E9 light-chain CDR1 : SGSIASNY (SEQ ID NO: 34) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDSSNLQWV (SEQ ID NO: 84) heavy-chain CDR1 : GYTFTSYG (SEQ ID NO: 127) heavy-chain CDR2: ISAYNGNT (SEQ ID NO: 138) heavy-chain CDR3: ARGIITMISDY (SEQ ID NO: 167);
3F1 light-chain CDR1 : GGSIADNF (SEQ ID NO: 45) light-chain CDR2: DYS (SEQ ID NO: 62) light-chain CDR3: QSYDISNPV (SEQ ID NO: 85) heavy-chain CDR1 : GFTFSSYG (SEQ ID NO: 128) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARDGGGGMDV (SEQ ID NO: 168);
3F11 light-chain CDR1 : HSDGRNY (SEQ ID NO: 46) light-chain CDR2: DTS (SEQ ID NO: 63) light-chain CDR3: HQFRRSVST (SEQ ID NO: 86) heavy-chain CDR1 : GYTFSSSG (SEQ ID NO: 129) heavy-chain CDR2: ISTYNGNT (SEQ ID NO: 146) heavy-chain CDR3: ATSIAVAGIDY (SEQ ID NO: 169);
3H7 light-chain CDR1 : TDSIASNY (SEQ ID NO: 47) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDGVNRGVI (SEQ ID NO: 87) heavy-chain CDR1 : GFTFSNYG (SEQ ID NO: 130) heavy-chain CDR2: IWHDGTNK (SEQ ID NO: 147) heavy-chain CDR3: VRDQGAGVWNGYSY (SEQ ID NO: 170);
3H11 light-chain CDR1 : SGNIASYY (SEQ ID NO: 48) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDRPNHVV (SEQ ID NO: 88)
heavy-chain CDR1 : GYTFTSYY (SEQ ID NO: 119) heavy-chain CDR2: INPSGGST (SEQ ID NO: 135) heavy-chain CDR3: ASSVVPAAIYDYYYGMDV (SEQ ID NO: 171);
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131 ) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172);
6F8 light-chain CDR1 : QTINNNY (SEQ ID NO: 50) light-chain CDR2: DAS (SEQ ID NO: 64) light-chain CDR3: QQYGSSPRT (SEQ ID NO: 90) heavy-chain CDR1 : GFTVSSNY (SEQ ID NO: 132) heavy-chain CDR2: IYSGGST (SEQ ID NO: 148) heavy-chain CDR3: ARDYGDYYFDY (SEQ ID NO: 173); or conservative sequence variants thereof.
14. The antigen-binding protein according to any one of the preceding claims, comprising a light-chain variable region comprising any one of SEQ ID NO: 1 -27 or a sequence sharing at least about 75% sequence similarity thereto; and/or a heavy-chain variable region comprising any one of SEQ ID NO: 91 -116 or a sequence sharing at least about 75% sequence similarity thereto.
15. A combination of two antigen-binding proteins, wherein a first antigen binding protein comprises the following CDR sequences:
5A6 light-chain CDR1 : QSISSY (SEQ ID NO: 49) light-chain CDR2: AAS (SEQ ID NO: 59) light-chain CDR3: QQSYNLPRT (SEQ ID NO: 89) heavy-chain CDR1 : GFTFSSYE (SEQ ID NO: 131) heavy-chain CDR2: ISYDGSNK (SEQ ID NO: 143) heavy-chain CDR3: ARLITMVRGEDY (SEQ ID NO: 172) and a second antigen-binding protein comprises the following CDR sequences:
3D1 1 light-chain CDR1 : SGNIASNY (SEQ ID NO: 44) light-chain CDR2: EDN (SEQ ID NO: 51 ) light-chain CDR3: QSYDNNIQV (SEQ ID NO: 83) heavy-chain CDR1 : GGSFSGYY (SEQ ID NO: 123) heavy-chain CDR2: INHSGST (SEQ ID NO: 140) heavy-chain CDR3: ARRWWLRGAFDI (SEQ ID NO: 166).
16. A pharmaceutical composition comprising the antigen-binding protein according to any one of claims 1 -14 or the combination according to claim 15, and a pharmaceutically acceptable carrier.
17. The antigen-binding protein according to any one of claims 1-14, the combination according to claim 15 or the pharmaceutical composition according to claim 16, for use in therapy, or as a diagnostic agent.
18. A nucleotide sequence encoding for the antigen-binding protein according to any one of claims 1 -14 or the combination according to claim 15.
19. The nucleotide sequence according to claim 18, wherein the nucleotide sequence comprises one of more of SEQ ID NO: 174-351 or a sequence sharing at least about 75% sequence identity thereto.
20. A bacteriophage, a host cell or a non-human organism comprising the nucleotide sequence according to claim 18 or claim 19.
21. A method of treating SARS-CoV-2 in a subject, the method comprising administering to the subject the antigen-binding protein according to any one of claims 1-14, the combination according to claim 15 or the pharmaceutical composition according to claim 16.
22. The method according to claim 21 , wherein where the SARS-CoV-2 comprises a V483A mutation, the method comprises administering to the subject the combination according to claim 15.
23. A method of detecting the presence of SARS-CoV-2 in a sample, the method comprising: contacting the sample with the antigen-binding protein according to any one of claims 1 -14; detecting a binding event involving the antigen-binding protein; wherein the detection of a binding event is indicative of the presence of
SARS-CoV-2 in the sample.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SG10202003204P | 2020-04-07 | ||
| SG10202003204P | 2020-04-07 | ||
| SG10202004828Q | 2020-05-22 | ||
| SG10202004828Q | 2020-05-22 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2021206636A1 true WO2021206636A1 (en) | 2021-10-14 |
Family
ID=78024068
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/SG2021/050195 WO2021206636A1 (en) | 2020-04-07 | 2021-04-07 | ANTIGEN-BINDING PROTEIN TO SARS-CoV-2 |
| PCT/US2021/026286 WO2021207433A2 (en) | 2020-04-07 | 2021-04-07 | Epitopes of sars-cov-2 neutralizing antibodies |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2021/026286 WO2021207433A2 (en) | 2020-04-07 | 2021-04-07 | Epitopes of sars-cov-2 neutralizing antibodies |
Country Status (1)
| Country | Link |
|---|---|
| WO (2) | WO2021206636A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114213531A (en) * | 2021-12-07 | 2022-03-22 | 中国人民解放军军事科学院军事医学研究院 | Novel coronavirus neutralizing antibody, antigen-binding fragment thereof and application thereof |
| RU2778942C1 (en) * | 2022-03-29 | 2022-08-29 | Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН) | RECOMBINANT SARS-CoV-2 S PROTEIN RBD-BINDING PROTEIN |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TW202241942A (en) * | 2020-12-21 | 2022-11-01 | 日商中外製藥股份有限公司 | Sars-cov-2 binding molecules and uses thereof |
| CN113999301B (en) * | 2021-12-07 | 2023-07-28 | 江苏中新医药有限公司 | anti-SARS-CoV-2 (COVID-19) S protein RBD monoclonal antibody |
| WO2024068777A1 (en) | 2022-09-28 | 2024-04-04 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Modified ace2 proteins with improved activity against sars-cov-2 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007044695A2 (en) * | 2005-10-07 | 2007-04-19 | Dana-Farber Cancer Institute | ANTIBODIES AGAINST SARS-CoV AND METHODS OF USE THEREOF |
| WO2010032059A2 (en) * | 2008-09-19 | 2010-03-25 | Medimmune Llc | Targeted binding agents directed to cd105 and uses thereof |
-
2021
- 2021-04-07 WO PCT/SG2021/050195 patent/WO2021206636A1/en active Application Filing
- 2021-04-07 WO PCT/US2021/026286 patent/WO2021207433A2/en active Application Filing
Non-Patent Citations (9)
| Title |
|---|
| CHUNYAN WANG, LI WENTAO, DRABEK DUBRAVKA, OKBA NISREEN M. A., VAN HAPEREN RIEN, OSTERHAUS ALBERT D. M. E., VAN KUPPEVELD FRANK J. : "A human monoclonal antibody blocking SARS-CoV-2 infection", NATURE COMMUNICATIONS, vol. 11, no. 1, 2020, XP055737066, DOI: 10.1038/s41467-020-16256-y * |
| JIANG S. ET AL.: "An emerging coronavirus causing pneumonia outbreak in Wuhan, China : calling for developing therapeutic and prophylactic strategies", EMERG MICROBES INFECT., vol. 9, no. 1, 31 January 2020 (2020-01-31), pages 275 - 277, XP055785642, [retrieved on 20210603], DOI: 10.1080/22221751.2020.1723441 * |
| JINDONG HAO, LI CHENGSHUN, LIN CHAO, HAO YANG, YU XIANFENG, XIA YIDAN, GAO FEI, JIANG ZIPING, WANG DONGXU: "Targeted point mutations of the m6 A modification in miR675 using RNA-guided base editing induce cell apoptosis", CELL DEATH AND DISEASE, vol. 40, no. 5, 29 May 2020 (2020-05-29), XP055720727, ISSN: 0144-8463, DOI: 10.1042/BSR20192933 * |
| LI Q. ET AL: "The Impact of Mutations in SARS-CoV-2 Spike on Viral Infectivity and Antigenicity", vol. 182, no. 5, 1 January 2020 (2020-01-01), pages 1284 - 1294, XP055824625, Retrieved from the Internet <URL:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7366990/pdf/main.pdf> [retrieved on 20210715] * |
| PARK TAMINA, LEE SANG-YEOP, KIM SEIL, KIM MI JEONG, KIM HONG GI, JUN SANGMI, KIM SEUNG IL, KIM BUM TAE, PARK EDMOND CHANGKYUN, PAR: "Abstract", BIORXIV, 27 February 2020 (2020-02-27), XP055812247, Retrieved from the Internet <URL:https://www.biorxiv.org/content/10.1101/2020.02.22.951178v1.full.pdf> [retrieved on 20210609], DOI: 10.1101/2020.02.22.951178 * |
| SUN Z. ET AL.: "Potent neutralization of SARS-CoV-2 by human antibody heavy- chain variable domains isolated from a large library with a new stable scaffold", MABS, vol. 12, no. 1, 16 June 2020 (2020-06-16), pages 1 - 6, XP055819835, [retrieved on 20210603], DOI: 10.1080/19420862.2020.1778435 * |
| WANG HONGYE, WU XIAN, ZHANG XIAOMEI, HOU XIN, LIANG TE, WANG DAN, TENG FEI, DAI JIAYU, DUAN HU, GUO SHUBIN, LI YONGZHE, YU XIAOBO: "SARS-CoV-2 Proteome Microarray for Mapping COVID-19 Antibody Interactions at Amino Acid Resolution", ACS CENTRAL SCIENCE, vol. 6, no. 12, 23 December 2020 (2020-12-23), pages 2238 - 2249, XP055864645, ISSN: 2374-7943, DOI: 10.1021/acscentsci.0c00742 * |
| WANG XIAOLI, GUO XIANGHUA, XIN QIANQIAN, PAN YANG, HU YALING, LI JING, CHU YANHUI, FENG YINGMEI, WANG QUANYI: "Neutralizing Antibody Responses to Severe Acute Respiratory Syndrome Coronavirus 2 in Coronavirus Disease 2019 Inpatients and Convalescent Patients", CLINICAL INFECTIOUS DISEASES, THE UNIVERSITY OF CHICAGO PRESS, CHICAGO, IL., US, vol. 71, no. 10, 4 June 2020 (2020-06-04), US , pages 2688 - 2694, XP055865066, ISSN: 1058-4838, DOI: 10.1093/cid/ciaa721 * |
| WU FAN, WANG AOJIE, LIU MEI, WANG QIMIN, CHEN JUN, XIA SHUAI, LING YUN, ZHANG YULING, XUN JINGNA, LU LU, JIANG SHIBO, LU HONGZHOU,: "Neutralizing antibody responses to SARS-CoV-2 in a COVID-19 recovered patient cohort and their implications", MEDRXIV, 20 April 2020 (2020-04-20), pages 1 - 20, XP055864866, Retrieved from the Internet <URL:https://www.medrxiv.org/content/10.1101/2020.03.30.20047365v2.full.pdf> [retrieved on 20211123], DOI: 10.1101/2020.03.30.20047365 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114213531A (en) * | 2021-12-07 | 2022-03-22 | 中国人民解放军军事科学院军事医学研究院 | Novel coronavirus neutralizing antibody, antigen-binding fragment thereof and application thereof |
| CN114213531B (en) * | 2021-12-07 | 2023-04-25 | 中国人民解放军军事科学院军事医学研究院 | Neutralizing antibodies against novel coronaviruses, antigen binding fragments thereof and uses thereof |
| RU2778942C1 (en) * | 2022-03-29 | 2022-08-29 | Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН) | RECOMBINANT SARS-CoV-2 S PROTEIN RBD-BINDING PROTEIN |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2021207433A2 (en) | 2021-10-14 |
| WO2021207433A3 (en) | 2021-11-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111690058B (en) | Antibodies with neutralizing activity against coronaviruses and uses thereof | |
| WO2021206636A1 (en) | ANTIGEN-BINDING PROTEIN TO SARS-CoV-2 | |
| WO2021180218A1 (en) | Monoclonal antibody against novel coronavirus and application thereof | |
| KR101732056B1 (en) | Neutralizing anti-influenza a virus antibodies and uses thereof | |
| CN112390879B (en) | Antibody targeting SARS-CoV-2 and its preparation method and use | |
| JP7034499B2 (en) | Uses of anti-FAM19A5 antibody for the treatment of fibrosis | |
| WO2021174595A1 (en) | Monoclonal antibody for resisting novel coronavirus and use thereof | |
| CN113388031B (en) | Monoclonal antibody 35B5, and preparation method and application thereof | |
| CN112625136A (en) | Bispecific antibodies having neutralizing activity against coronaviruses and uses thereof | |
| JP2016503817A (en) | Anti-granulinic antibodies and methods of use thereof | |
| CN113388030B (en) | Monoclonal antibody 32C7, and preparation method and application thereof | |
| WO2021212049A2 (en) | Anti-sars-cov-2 monoclonal antibodies | |
| WO2021174594A1 (en) | Anti-novel coronavirus monoclonal antibody and application thereof | |
| Errico et al. | Structural mechanism of SARS-CoV-2 neutralization by two murine antibodies targeting the RBD | |
| KR20220012826A (en) | SARS-CoV-2 NEUTRALIZING BINDING MOLECULE BINDING TO EPITOPES OF SARS-CoV-2 SPIKE PROTEIN | |
| WO2021237516A1 (en) | Sars-cov-2 antibody and application thereof | |
| WO2021218947A1 (en) | Anti-novel coronavirus monoclonal antibody and application thereof | |
| JP2024518335A (en) | Human neutralizing monoclonal antibodies against SARS-CoV-2 and their uses | |
| WO2021229561A1 (en) | Anti-sars-cov-2 antibodies | |
| WO2022095996A1 (en) | Anti-sars-cov-2 antibody and application thereof | |
| WO2023199943A1 (en) | Antibody used to treat coronavirus infection | |
| EP4088782A1 (en) | Antibodies | |
| KR20220139244A (en) | Composition Comprising Two or more SARS-CoV-2 Virus Neutralizing Binding Molecules | |
| WO2023148641A1 (en) | Monoclonal antibodies specific to receptor binding domain of sars-cov2 and uses thereof | |
| CN116867801A (en) | Single domain antibodies that bind to SARS-CoV-2 spike protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 21784547 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 21784547 Country of ref document: EP Kind code of ref document: A1 |