JP6925336B2 - 抗呼吸器合胞体ウイルスの完全ヒト抗体 - Google Patents
抗呼吸器合胞体ウイルスの完全ヒト抗体 Download PDFInfo
- Publication number
- JP6925336B2 JP6925336B2 JP2018527811A JP2018527811A JP6925336B2 JP 6925336 B2 JP6925336 B2 JP 6925336B2 JP 2018527811 A JP2018527811 A JP 2018527811A JP 2018527811 A JP2018527811 A JP 2018527811A JP 6925336 B2 JP6925336 B2 JP 6925336B2
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- antigen
- seq
- nucleic acid
- rsv
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000700605 Viruses Species 0.000 title description 7
- 230000003097 anti-respiratory effect Effects 0.000 title description 3
- 239000012634 fragment Substances 0.000 claims description 28
- 239000000427 antigen Substances 0.000 claims description 25
- 108091007433 antigens Proteins 0.000 claims description 25
- 102000036639 antigens Human genes 0.000 claims description 25
- 150000007523 nucleic acids Chemical class 0.000 claims description 22
- 239000013598 vector Substances 0.000 claims description 20
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 18
- 108020004707 nucleic acids Proteins 0.000 claims description 17
- 102000039446 nucleic acids Human genes 0.000 claims description 17
- 206010006448 Bronchiolitis Diseases 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 108020001507 fusion proteins Proteins 0.000 claims description 5
- 102000037865 fusion proteins Human genes 0.000 claims description 5
- 206010035737 Pneumonia viral Diseases 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 4
- 208000009421 viral pneumonia Diseases 0.000 claims description 4
- 208000029523 Interstitial Lung disease Diseases 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 2
- 108010003723 Single-Domain Antibodies Proteins 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 229940124597 therapeutic agent Drugs 0.000 claims description 2
- 241000724205 Rice stripe tenuivirus Species 0.000 claims 4
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 28
- 239000000872 buffer Substances 0.000 description 10
- 239000013612 plasmid Substances 0.000 description 10
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 229940036185 synagis Drugs 0.000 description 7
- 238000002965 ELISA Methods 0.000 description 6
- 239000013604 expression vector Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- 108090000288 Glycoproteins Proteins 0.000 description 4
- 102000003886 Glycoproteins Human genes 0.000 description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 206010035664 Pneumonia Diseases 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000011535 reaction buffer Substances 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 241000725643 Respiratory syncytial virus Species 0.000 description 2
- 206010061603 Respiratory syncytial virus infection Diseases 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- KLOHDWPABZXLGI-YWUHCJSESA-M ampicillin sodium Chemical compound [Na+].C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C([O-])=O)(C)C)=CC=CC=C1 KLOHDWPABZXLGI-YWUHCJSESA-M 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 210000004700 fetal blood Anatomy 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 108010068327 4-hydroxyphenylpyruvate dioxygenase Proteins 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108091006027 G proteins Proteins 0.000 description 1
- 102000030782 GTP binding Human genes 0.000 description 1
- 108091000058 GTP-Binding Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- 101710180643 Leishmanolysin Proteins 0.000 description 1
- 101710085938 Matrix protein Proteins 0.000 description 1
- 201000005505 Measles Diseases 0.000 description 1
- 108010090054 Membrane Glycoproteins Proteins 0.000 description 1
- 102000012750 Membrane Glycoproteins Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000011931 Nucleoproteins Human genes 0.000 description 1
- 108010061100 Nucleoproteins Proteins 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 241000711504 Paramyxoviridae Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 206010036590 Premature baby Diseases 0.000 description 1
- 101710132653 Protein M2 Proteins 0.000 description 1
- 108010059722 Viral Fusion Proteins Proteins 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 229960001931 ampicillin sodium Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000002497 edematous effect Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 238000011577 humanized mouse model Methods 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000001739 intranuclear inclusion body Anatomy 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229960000402 palivizumab Drugs 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 210000004180 plasmocyte Anatomy 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 210000002536 stromal cell Anatomy 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000011830 transgenic mouse model Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Images
Classifications
-
- 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
- C07K16/1027—Paramyxoviridae, e.g. respiratory syncytial virus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/295—Polyvalent viral antigens; Mixtures of viral and bacterial antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6075—Viral proteins
-
- 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
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/115—Paramyxoviridae, e.g. parainfluenza virus
- G01N2333/135—Respiratory syncytial virus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Physics & Mathematics (AREA)
- Pulmonology (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Plant Pathology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Cell Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Food Science & Technology (AREA)
Description
配列番号:6、配列番号:8、及び配列番号:10に示されるアミノ酸配列から選択されるアミノ酸配列を有する少なくとも1本の重鎖CDR、及び/又は
配列番号:12、配列番号:14、及び配列番号:16に示されるアミノ酸配列から選択されるアミノ酸配列を有する少なくとも1本の軽鎖CDRを含有する。
配列番号:6、配列番号:8、及び配列番号:10に示されるアミノ酸配列から選択されるアミノ酸配列を有する3本の重鎖CDR、及び/又は
配列番号:12、配列番号:14、及び配列番号:16に示されるアミノ酸配列から選択されるアミノ酸配列を有する3本の軽鎖CDRを含有する。
配列番号:3に示されるアミノ酸配列を有する重鎖可変領域及び/又は配列番号:4に示されるアミノ酸配列を有する軽鎖可変領域を含み、上記抗体は、抗RSV完全ヒト化モノクローナル抗体R66である。
重鎖CDRをコードする、配列番号:5、配列番号:7、及び配列番号:9から選択される少なくとも1つのヌクレオチド配列、及び/又は
軽鎖CDRをコードする、配列番号:11、配列番号:13、及び配列番号:15から選択される少なくとも1つのヌクレオチド配列を含有する。
配列番号:1に示される、重鎖可変領域をコードする核酸、及び/又は配列番号:2に示される、軽鎖可変領域をコードする核酸を含有する。
ステップ(1)において、発現ベクターを提供し、上記発現ベクターは、本発明の抗RSV抗体(好ましくは、抗RSV完全ヒト化モノクローナル抗体)をコードするDNA分子、及び上記DNA分子と作動可能に連結される発現調節配列を含有する。
ステップ(2)において、上記発現ベクターで宿主細胞を形質転換する。
ステップ(3)において、上記抗RSV抗体(好ましくは、抗RSV完全ヒト化モノクローナル抗体)の発現に適した条件下で上記宿主細胞を培養する。
ステップ(4)において、分離精製して上記抗RSV抗体(好ましくは、抗RSV完全ヒト化モノクローナル抗体)を得る。
1、R66の重鎖可変領域及び軽鎖可変領域の人工合成
配列が配列番号:1に示すようなモノクローナル抗体R66の重鎖可変領域、配列が配列番号:2に示すようなモノクローナル抗体R66の軽鎖(kappa)可変領域の核酸配列をGENEWIZ社に人工合成してもらった。
高速DNA産物精製キット(康為世紀から購入)によりPCR産物を回収して40μlのPCR産物を得た。
実施例1で得られたR66の重鎖(RH66)を含有するプラスミド及びR66の軽鎖(RK66)を含有するプラスミドにより、それぞれ293T細胞をトランスフェクションした。まず、Opti−MEM(1X)bufferでそれぞれプラスミド及びPEIを希釈し、そしてPEI−Opti−MEM混合物をプラスミド−Opti−MEM混合物管にゆっくりと加え、室温で20分間静置した後、PEIとプラスミドの混合物を細胞懸濁液に加えた。トランスフェクション時の細胞濃度は0.25〜0.5×106細胞/mlであり、細胞のトランスフェクションには、1ウェルあたり、R66重鎖を含有する2.5μgのプラスミド+R66軽鎖を含有する2.5μgのプラスミド+10μgのPEIを用いた。トランスフェクション終了後、37℃で48h培養した後、上澄みを収集し、ELISAで上澄みを検出した。
使用した試薬
PBS(1X):PBS(10X)及び脱イオン水で調製される。
PBST:PBS(1X)に最終濃度が0.05%となるまでTween−20を加える。
ブロッキング液:PBS(1X)+2%BSA+2%新生仔ウシ血清;使用直前に調製。
希釈液:PBST+1%BSA;抗体希釈用。
停止液:6ml 95%−98%の濃硫酸を180ml水にゆっくりと加え、冷却しておく。
一次抗体:本発明により精製されたR66抗体を作業濃度が10μg/mlになるように希釈した。
二次抗体:Peroxidase−conjugated AffiniPure Goat Anti−Human IgG(H+L)(Jackson Immuno Researchから購入);1.5mlのRNaseフリーの水で完全に溶解しておく。
抗体可変領域の配列情報をIgBLASTプログラム(version 1.6.1)に導入し、ヒト可変領域のオリジナルの配列ライブラリーと比較分析し、抗体の可変領域から3つのフレームワーク領域(FR)及び2つの相補性決定領域(CDR)を画定した。その後、抗体配列をIMGT High V−Questシステムに導入し、上記と同じライブラリーを用いて比較してCDR3及びFR4を確定した。なお、すべての抗体配列の番号付けはKABATシステムに基づく。結果を図4に示す。
Claims (12)
- 抗RSV抗体又はその抗原結合断片であって、前記抗体は、
配列番号:3に示されるアミノ酸配列を有する重鎖可変領域、及び
配列番号:4に示されるアミノ酸配列を有する軽鎖可変領域を含有する、完全ヒト化抗体又はその抗原結合断片。 - 前記抗原結合断片は、Fab、Fab’、F(ab’) 2、scFv、Fv、dsFv、二重抗体、Fd及びFd’断片から選択される請求項1に記載の完全ヒト化抗体又はその抗原結合断片。
- 請求項1または2に記載の完全ヒト化抗体の単一重鎖、単一軽鎖又はその抗原結合断片を有するシングルドメイン抗体、抗体融合タンパク質抗体、抗体/抗体断片−因子融合タンパク質、又は抗体/抗体断片−化学複合体。
- 請求項1または2に記載の完全ヒト化抗体又はその抗原結合断片をコードする核酸。
- 配列番号:1に示される、重鎖可変領域をコードする核酸、及び
配列番号:2に示される、軽鎖可変領域をコードする核酸を有する、請求項4に記載の核酸。 - 請求項4または5に記載の核酸を有するベクター。
- 請求項4または5に記載の核酸、又は請求項6に記載のベクターを有する宿主細胞。
- 請求項6に記載のベクターで形質転換されたものである請求項7に記載の宿主細胞。
- 請求項1または2に記載の完全ヒト化抗体又はその抗原結合断片、請求項3に記載の抗体、融合タンパク質又は複合体、請求項4または5に記載の核酸、請求項6に記載のベクター、或いは請求項7若しくは請求項8に記載の宿主細胞の、RSV関連疾患を予防又は治療するための治療剤の調製における使用。
- 前記RSV関連疾患は、ウイルス性肺炎、間質性肺炎、細気管支炎からなる群より選択される請求項9に記載の使用。
- 請求項1または2に記載の完全ヒト化抗体又はその抗原結合断片、請求項4または5に記載の核酸、請求項6に記載のベクター、又は請求項7若しくは請求項8に記載の宿主細胞の、RSVを検出するための検出試薬の調製における使用。
- 治療有効量に、請求項1または2に記載の完全ヒト化抗体又はその抗原結合断片、請求項4または5に記載の核酸、請求項6に記載のベクター、又は請求項7若しくは請求項8に記載の宿主細胞、及び1種又は複数種の薬学的に許容されるベクター又は賦形剤を含有する、RSV関連疾患を治療するための医薬組成物。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510867051.9 | 2015-11-30 | ||
CN201510867051 | 2015-11-30 | ||
CN201611036424.9A CN106496324B (zh) | 2015-11-30 | 2016-11-23 | 一种抗呼吸道合胞病毒的全人源抗体 |
CN201611036424.9 | 2016-11-23 | ||
PCT/CN2016/107658 WO2017092645A1 (zh) | 2015-11-30 | 2016-11-29 | 一种抗呼吸道合胞病毒的全人源抗体 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2019507103A JP2019507103A (ja) | 2019-03-14 |
JP6925336B2 true JP6925336B2 (ja) | 2021-08-25 |
Family
ID=58327667
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2018527811A Active JP6925336B2 (ja) | 2015-11-30 | 2016-11-29 | 抗呼吸器合胞体ウイルスの完全ヒト抗体 |
Country Status (7)
Country | Link |
---|---|
US (1) | US10149903B2 (ja) |
EP (1) | EP3385278A1 (ja) |
JP (1) | JP6925336B2 (ja) |
KR (1) | KR102184151B1 (ja) |
CN (1) | CN106496324B (ja) |
AU (1) | AU2016363455B2 (ja) |
WO (1) | WO2017092645A1 (ja) |
Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106496324B (zh) * | 2015-11-30 | 2020-01-14 | 天津昊免生物技术有限公司 | 一种抗呼吸道合胞病毒的全人源抗体 |
CN111606992B (zh) * | 2019-02-25 | 2022-08-19 | 天津昊免生物技术有限公司 | 一种抗呼吸道合胞病毒的全人源抗体 |
CN111606993B (zh) * | 2019-02-26 | 2022-06-28 | 中国科学院上海生命科学研究院 | 抗呼吸道合胞病毒的全人广谱中和抗体4f1及其应用 |
CN112239498B (zh) * | 2019-07-16 | 2022-08-19 | 天津昊免生物技术有限公司 | 一种抗丙型肝炎病毒抗体 |
CN112239497B (zh) * | 2019-07-16 | 2022-09-13 | 天津昊免生物技术有限公司 | 一种抗丙型肝炎病毒抗体 |
CN114805561B (zh) * | 2019-08-02 | 2023-07-18 | 苏州高泓利康生物科技有限公司 | 针对呼吸道合胞病毒的蛋白结合分子 |
CN112225803B (zh) * | 2020-10-20 | 2022-08-05 | 长春百克生物科技股份公司 | 纳米抗体及其应用 |
US20240059763A1 (en) * | 2020-12-18 | 2024-02-22 | Zhuhai Trinomab Pharmaceutical Co., Ltd. | Respiratory syncytial virus-specific binding molecule |
CN115466326B (zh) * | 2021-06-11 | 2024-06-04 | 中国科学院微生物研究所 | 一种呼吸道合胞病毒的人源单克隆抗体及其应用 |
CN114456264B (zh) * | 2022-03-24 | 2023-02-03 | 中国科学院微生物研究所 | 一种新型冠状病毒稀有广谱表位的人源抗体及其应用 |
CN116987183A (zh) * | 2022-07-22 | 2023-11-03 | 北京智仁美博生物科技有限公司 | 抗呼吸道合胞病毒中和性抗体及其用途 |
CN116063468A (zh) * | 2022-08-30 | 2023-05-05 | 武汉班科生物技术有限公司 | 中和呼吸道合胞病毒的c-型单域抗体及应用 |
WO2024120517A1 (zh) * | 2022-12-08 | 2024-06-13 | 南京诺唯赞生物科技股份有限公司 | 特异性结合rsv的抗体 |
CN117310163B (zh) * | 2023-11-23 | 2024-02-13 | 北京贝思泰生物科技有限公司 | 一种基于重组抗体的呼吸道合胞病毒检测方法 |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050175986A1 (en) * | 2000-05-09 | 2005-08-11 | Smit Kline Beecham Corporation | Human monoclonal antibody |
CN1986569A (zh) * | 2005-12-23 | 2007-06-27 | 首都儿科研究所 | 人源抗呼吸道合胞病毒中和性基因工程Fab抗体 |
MX2010004482A (es) * | 2007-10-25 | 2010-07-06 | Trellis Bioscience Inc | Anticuerpos anti-proteina g vsr. |
ES2553440T3 (es) * | 2009-08-13 | 2015-12-09 | Crucell Holland B.V. | Anticuerpos contra el virus sincitial respiratorio humano (VSR) y método de uso |
AU2009353693B2 (en) * | 2009-10-06 | 2016-07-21 | Medimmune Ltd | RSV-specific binding molecule |
CA3062786C (en) * | 2010-07-09 | 2022-04-19 | Janssen Vaccines & Prevention B.V. | Anti-human respiratory syncytial virus (rsv) antibodies and methods of use |
US9718876B2 (en) * | 2011-11-17 | 2017-08-01 | Aimm Therapeutics B.V. | RSV G protein specific antibodies |
TWI659968B (zh) * | 2013-03-14 | 2019-05-21 | 再生元醫藥公司 | 針對呼吸道融合病毒f蛋白質的人類抗體及其使用方法 |
CA2906960C (en) * | 2013-03-15 | 2021-07-20 | Xiamen University | Epitope of rsv fusion protein and antibody recognizing the epitope |
US10035842B2 (en) * | 2013-04-15 | 2018-07-31 | Janssen Vaccines & Prevention B.V. | Human antibodies binding to RSV G proteins |
ES2706759T3 (es) | 2013-07-10 | 2019-04-01 | Tepha Inc | Anclajes de sutura blandos |
CN106496324B (zh) * | 2015-11-30 | 2020-01-14 | 天津昊免生物技术有限公司 | 一种抗呼吸道合胞病毒的全人源抗体 |
CN109232737A (zh) * | 2016-03-01 | 2019-01-18 | 苏州博泰神州生物技术有限公司 | 一种针对rsv粘附g蛋白表面抗原的全人源单克隆抗体 |
-
2016
- 2016-11-23 CN CN201611036424.9A patent/CN106496324B/zh active Active
- 2016-11-29 AU AU2016363455A patent/AU2016363455B2/en active Active
- 2016-11-29 WO PCT/CN2016/107658 patent/WO2017092645A1/zh active Application Filing
- 2016-11-29 EP EP16869955.1A patent/EP3385278A1/en active Pending
- 2016-11-29 KR KR1020187017477A patent/KR102184151B1/ko active IP Right Grant
- 2016-11-29 JP JP2018527811A patent/JP6925336B2/ja active Active
-
2018
- 2018-05-29 US US15/990,999 patent/US10149903B2/en active Active
Also Published As
Publication number | Publication date |
---|---|
AU2016363455B2 (en) | 2019-07-11 |
US20180264103A1 (en) | 2018-09-20 |
JP2019507103A (ja) | 2019-03-14 |
KR102184151B1 (ko) | 2020-11-30 |
EP3385278A4 (en) | 2018-10-10 |
KR20180089438A (ko) | 2018-08-08 |
WO2017092645A1 (zh) | 2017-06-08 |
EP3385278A1 (en) | 2018-10-10 |
CN106496324A (zh) | 2017-03-15 |
US10149903B2 (en) | 2018-12-11 |
CN106496324B (zh) | 2020-01-14 |
AU2016363455A1 (en) | 2018-07-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6925336B2 (ja) | 抗呼吸器合胞体ウイルスの完全ヒト抗体 | |
CN112250763B (zh) | 靶向SARS-CoV-2冠状病毒的抗体及其诊断和检测用途 | |
CN111690058A (zh) | 针对冠状病毒的具有中和活性的抗体及其用途 | |
US10196438B2 (en) | Human antibodies binding to RSV G protein | |
CA2908921C (en) | Human antibodies binding to rsv g protein | |
CN110642944B (zh) | 一种中和人感染h7n9甲型流感病毒的抗体及其用途 | |
CN111333723B (zh) | 针对狂犬病病毒g蛋白的单克隆抗体及其用途 | |
CN116514960A (zh) | 一种呼吸道合胞病毒的全人源单克隆抗体及其应用 | |
CN110437333B (zh) | Sftsv抑制剂及其应用 | |
AU2016277887B2 (en) | Immunoglobulin single variable domain antibody against RSV prefusion F protein | |
WO2022258068A1 (zh) | 一种呼吸道合胞病毒的抗体及其应用 | |
US20160200799A1 (en) | Anti-influenza virus neutralizing antibody | |
CN111606992B (zh) | 一种抗呼吸道合胞病毒的全人源抗体 | |
BR102012011493A2 (pt) | Método de produção e obtenção de fragmento fab do anticorpo anti-digoxina monoclonal a partir da técnica de clonagem em biologia molecular | |
WO2022143815A1 (zh) | 抗新冠病毒受体结合区域表位中和抗体及其应用 | |
CN116606373A (zh) | 新型冠状病毒中和抗体及其用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20180523 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20180523 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20190508 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20190703 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20191002 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200120 |
|
C60 | Trial request (containing other claim documents, opposition documents) |
Free format text: JAPANESE INTERMEDIATE CODE: C60 Effective date: 20200120 |
|
A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20200127 |
|
C21 | Notice of transfer of a case for reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C21 Effective date: 20200128 |
|
A912 | Re-examination (zenchi) completed and case transferred to appeal board |
Free format text: JAPANESE INTERMEDIATE CODE: A912 Effective date: 20200327 |
|
C211 | Notice of termination of reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C211 Effective date: 20200331 |
|
C22 | Notice of designation (change) of administrative judge |
Free format text: JAPANESE INTERMEDIATE CODE: C22 Effective date: 20201020 |
|
C13 | Notice of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: C13 Effective date: 20210302 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210510 |
|
C23 | Notice of termination of proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C23 Effective date: 20210601 |
|
C03 | Trial/appeal decision taken |
Free format text: JAPANESE INTERMEDIATE CODE: C03 Effective date: 20210706 |
|
C30A | Notification sent |
Free format text: JAPANESE INTERMEDIATE CODE: C3012 Effective date: 20210706 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20210803 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6925336 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |