WO2022105922A1 - Peptides courts dérivés d'un antigène ssx2 - Google Patents
Peptides courts dérivés d'un antigène ssx2 Download PDFInfo
- Publication number
- WO2022105922A1 WO2022105922A1 PCT/CN2021/132192 CN2021132192W WO2022105922A1 WO 2022105922 A1 WO2022105922 A1 WO 2022105922A1 CN 2021132192 W CN2021132192 W CN 2021132192W WO 2022105922 A1 WO2022105922 A1 WO 2022105922A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- peptide
- cell
- molecule
- pmhc complex
- present
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 151
- 239000000427 antigen Substances 0.000 title claims abstract description 28
- 108091007433 antigens Proteins 0.000 title claims abstract description 28
- 102000036639 antigens Human genes 0.000 title claims abstract description 28
- 102000004196 processed proteins & peptides Human genes 0.000 title abstract description 75
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 27
- 210000004027 cell Anatomy 0.000 claims description 79
- 108091008874 T cell receptors Proteins 0.000 claims description 52
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 claims description 44
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 35
- 230000027455 binding Effects 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 31
- 150000007523 nucleic acids Chemical class 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 18
- 239000013598 vector Substances 0.000 claims description 17
- 150000001413 amino acids Chemical class 0.000 claims description 16
- 108020004707 nucleic acids Proteins 0.000 claims description 16
- 102000039446 nucleic acids Human genes 0.000 claims description 16
- 239000012634 fragment Substances 0.000 claims description 14
- 201000011510 cancer Diseases 0.000 claims description 12
- 108010075704 HLA-A Antigens Proteins 0.000 claims description 10
- 229960005486 vaccine Drugs 0.000 claims description 9
- 239000003814 drug Substances 0.000 claims description 8
- 239000008194 pharmaceutical composition Substances 0.000 claims description 8
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 4
- 238000000338 in vitro Methods 0.000 claims description 4
- 239000003937 drug carrier Substances 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 2
- 230000000295 complement effect Effects 0.000 claims description 2
- 230000000638 stimulation Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 3
- 102000011786 HLA-A Antigens Human genes 0.000 claims 2
- 101000880770 Homo sapiens Protein SSX2 Proteins 0.000 abstract description 20
- 102100037686 Protein SSX2 Human genes 0.000 abstract description 20
- 229920001184 polypeptide Polymers 0.000 description 18
- 210000004881 tumor cell Anatomy 0.000 description 15
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical group N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 10
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 9
- 102100028972 HLA class I histocompatibility antigen, A alpha chain Human genes 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 239000012071 phase Substances 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 230000009258 tissue cross reactivity Effects 0.000 description 8
- 238000003114 enzyme-linked immunosorbent spot assay Methods 0.000 description 7
- 230000014509 gene expression Effects 0.000 description 7
- 210000003000 inclusion body Anatomy 0.000 description 7
- 241000588724 Escherichia coli Species 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 108060003951 Immunoglobulin Proteins 0.000 description 5
- 108010090804 Streptavidin Proteins 0.000 description 5
- 239000007983 Tris buffer Substances 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000006287 biotinylation Effects 0.000 description 5
- 238000007413 biotinylation Methods 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 102000018358 immunoglobulin Human genes 0.000 description 5
- 238000004949 mass spectrometry Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 229940124597 therapeutic agent Drugs 0.000 description 5
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 235000020958 biotin Nutrition 0.000 description 4
- 239000011616 biotin Substances 0.000 description 4
- 229960002685 biotin Drugs 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000004153 renaturation Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000011510 Elispot assay Methods 0.000 description 3
- 102000008949 Histocompatibility Antigens Class I Human genes 0.000 description 3
- 108010088652 Histocompatibility Antigens Class I Proteins 0.000 description 3
- 102000007079 Peptide Fragments Human genes 0.000 description 3
- 108010033276 Peptide Fragments Proteins 0.000 description 3
- 210000000612 antigen-presenting cell Anatomy 0.000 description 3
- 239000011324 bead Substances 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 210000004443 dendritic cell Anatomy 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 239000000032 diagnostic agent Substances 0.000 description 3
- 229940039227 diagnostic agent Drugs 0.000 description 3
- 230000013595 glycosylation Effects 0.000 description 3
- 238000006206 glycosylation reaction Methods 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000002998 immunogenetic effect Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000004481 post-translational protein modification Effects 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 208000023275 Autoimmune disease Diseases 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- 102000001253 Protein Kinase Human genes 0.000 description 2
- 206010039491 Sarcoma Diseases 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000021736 acetylation Effects 0.000 description 2
- 238000006640 acetylation reaction Methods 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 230000001086 cytosolic effect Effects 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 229940072221 immunoglobulins Drugs 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- -1 pMHC complexes Proteins 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 238000002823 phage display Methods 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108060006633 protein kinase Proteins 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000010532 solid phase synthesis reaction Methods 0.000 description 2
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-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
- PMUNIMVZCACZBB-UHFFFAOYSA-N 2-hydroxyethylazanium;chloride Chemical compound Cl.NCCO PMUNIMVZCACZBB-UHFFFAOYSA-N 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 150000008574 D-amino acids Chemical class 0.000 description 1
- 235000000638 D-biotin Nutrition 0.000 description 1
- 239000011665 D-biotin Substances 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 108700034637 EC 3.2.-.- Proteins 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 108010053070 Glutathione Disulfide Proteins 0.000 description 1
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 description 1
- 108010027412 Histocompatibility Antigens Class II Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- FSVCELGFZIQNCK-UHFFFAOYSA-N N,N-bis(2-hydroxyethyl)glycine Chemical compound OCCN(CCO)CC(O)=O FSVCELGFZIQNCK-UHFFFAOYSA-N 0.000 description 1
- BZQFBWGGLXLEPQ-UHFFFAOYSA-N O-phosphoryl-L-serine Natural products OC(=O)C(N)COP(O)(O)=O BZQFBWGGLXLEPQ-UHFFFAOYSA-N 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 102000042330 SSX family Human genes 0.000 description 1
- 108091077753 SSX family Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 208000000260 Warts Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000007998 bicine buffer Substances 0.000 description 1
- 230000008512 biological response Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000021523 carboxylation Effects 0.000 description 1
- 238000006473 carboxylation reaction Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 210000004671 cell-free system Anatomy 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 238000005094 computer simulation Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000006240 deamidation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229950006137 dexfosfoserine Drugs 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000002825 functional assay Methods 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 238000011223 gene expression profiling Methods 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- YPZRWBKMTBYPTK-BJDJZHNGSA-N glutathione disulfide Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@H](C(=O)NCC(O)=O)CSSC[C@@H](C(=O)NCC(O)=O)NC(=O)CC[C@H](N)C(O)=O YPZRWBKMTBYPTK-BJDJZHNGSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 208000014829 head and neck neoplasm Diseases 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 239000012145 high-salt buffer Substances 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 230000001024 immunotherapeutic effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 239000002062 molecular scaffold Substances 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 238000010844 nanoflow liquid chromatography Methods 0.000 description 1
- 238000002170 nanoflow liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- YPZRWBKMTBYPTK-UHFFFAOYSA-N oxidized gamma-L-glutamyl-L-cysteinylglycine Natural products OC(=O)C(N)CCC(=O)NC(C(=O)NCC(O)=O)CSSCC(C(=O)NCC(O)=O)NC(=O)CCC(N)C(O)=O YPZRWBKMTBYPTK-UHFFFAOYSA-N 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- BZQFBWGGLXLEPQ-REOHCLBHSA-N phosphoserine Chemical compound OC(=O)[C@@H](N)COP(O)(O)=O BZQFBWGGLXLEPQ-REOHCLBHSA-N 0.000 description 1
- USRGIUJOYOXOQJ-GBXIJSLDSA-N phosphothreonine Chemical compound OP(=O)(O)O[C@H](C)[C@H](N)C(O)=O USRGIUJOYOXOQJ-GBXIJSLDSA-N 0.000 description 1
- DCWXELXMIBXGTH-UHFFFAOYSA-N phosphotyrosine Chemical compound OC(=O)C(N)CC1=CC=C(OP(O)(O)=O)C=C1 DCWXELXMIBXGTH-UHFFFAOYSA-N 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000002708 random mutagenesis Methods 0.000 description 1
- 238000010223 real-time analysis Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 201000010153 skin papilloma Diseases 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 206010041823 squamous cell carcinoma Diseases 0.000 description 1
- 208000017572 squamous cell neoplasm Diseases 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 1
- 206010042863 synovial sarcoma Diseases 0.000 description 1
- 108010042703 synovial sarcoma X breakpoint proteins Proteins 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 230000002381 testicular Effects 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4748—Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- 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
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001184—Cancer testis antigens, e.g. SSX, BAGE, GAGE or SAGE
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464484—Cancer testis antigens, e.g. SSX, BAGE, GAGE or SAGE
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70503—Immunoglobulin superfamily
- C07K14/7051—T-cell receptor (TcR)-CD3 complex
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70503—Immunoglobulin superfamily
- C07K14/70539—MHC-molecules, e.g. HLA-molecules
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
-
- 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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
-
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
-
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0636—T lymphocytes
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
-
- 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
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
- G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
- G01N2333/7051—T-cell receptor (TcR)-CD3 complex
Definitions
- the present invention relates to short peptides derived from SSX2 antigen, in particular, to newly discovered short peptides derived from tumor antigen SSX2, complexes formed by the short peptides with MHC molecules, and uses of the short peptides and complexes.
- the present invention also relates to molecules that bind to the above-mentioned short peptides or complexes, and uses of these molecules.
- markers of a pathological or abnormal state can be used not only as markers for disease diagnosis, but also for the production of diagnostic and/or therapeutic agents.
- markers of cancer are used to generate specific antibodies.
- these molecules can also effectively stimulate the specific immune response of cytotoxic T lymphocytes (CTL) and exert anti-tumor efficacy.
- CTL cytotoxic T lymphocytes
- TCR T cell receptors that can bind to the "marker” through activated CTL. (TCR) as a therapeutic agent. Therefore, these molecules play a very important role in the diagnosis and treatment of related diseases.
- tumor antigens are proteolytically processed into polypeptide fragments of 8-16 amino acids in length, that is, CTL epitopes, which in turn interact with the major histocompatibility complex (MHC, CTL) in the lumen of the endoplasmic reticulum.
- MHC major histocompatibility complex
- Human MHC is usually referred to as HLA gene or HLA complex
- HLA gene or HLA complex Human MHC molecules combine to form peptide-MHC complex (peptide-MHC complex, pMHC), and finally present pMHC to the cell surface for recognition by TCR on the surface of CD8 + T cells.
- peptide-MHC complex peptide-MHC complex, pMHC
- pMHC peptide-MHC complex
- the discovery and determination of the presented polypeptide fragments is a complex process, because the presentation of polypeptides by HLA is the result of the enzymatic hydrolysis of the antigenic protein and the interaction of the polypeptide fragments with HLA. This shows that the complete tumor antigen molecule cannot provide any information for the discovery and identification of polypeptide fragments.
- Many literatures have published methods using computer simulation, such as the public databases SYFPEITHI (Rammensee, et al., Immunogenetics. 1999(50): 213-219) and BIMAS (Parker, et al., J. Immunol. 1994. 152: 163), Predictive algorithms are provided to identify which polypeptide fragments are likely to be presented.
- SSX2 is a synovial sarcoma X breakpoint, also known as HOM-MEL-40.
- SSX2 is one of ten highly homologous nucleic acid proteins of the SSX family.
- SSX protein is a tumor testis antigen and is only expressed in tumor cells and testicular blasts without MHC expression.
- SSX2 is expressed in a variety of human cancer cells including, but not limited to, melanoma, head and neck cancer, lymphoma, various myelomas, pancreatic cancer, prostate cancer, sarcoma, hepatocellular carcinoma, and colon cancer.
- peptides derived from SSX2 as targets for the above-mentioned cancers, can not only be used as markers for the diagnosis of the above-mentioned diseases, but also can be used to generate prophylactic and/or therapeutic agents for the above-mentioned diseases, such as antibodies or T cell receptors.
- the present invention utilizes mass spectrometer analysis and identification to discover for the first time a polypeptide fragment derived from tumor antigen SSX2 presented on the surface of tumor cells.
- the purpose of the present invention is to provide a newly discovered short peptide derived from tumor antigen SSX2, the complex formed by the short peptide and MHC molecules, and the use of the short peptide and the complex.
- a first aspect of the present invention provides a short peptide derived from the SSX2 antigen, the peptide comprising the amino acid sequence: AQIPEKIQK (SEQ ID NO: 1);
- the peptide can form a complex with MHC molecules.
- the peptide consists of 9 or 10 amino acids.
- the peptide consists of 9 amino acids.
- amino acid sequence of the peptide is SEQ ID NO: 1.
- the second aspect of the present invention provides a pMHC complex comprising the peptide of the first aspect of the present invention.
- amino acid sequence of the peptide in the pMHC complex is SEQ ID NO: 1.
- the type of MHC molecule is HLA-A*11.
- the type of MHC molecule is HLA-A*1101.
- the pMHC complex is a multimer.
- the pMHC complex is soluble.
- the pMHC complex is biotinylated.
- the third aspect of the present invention provides an isolated cell, the cell surface presents the pMHC complex of the second aspect of the present invention.
- the fourth aspect of the present invention provides a nucleic acid molecule comprising a nucleic acid sequence encoding the peptide of the first aspect of the present invention or a complementary sequence thereof.
- the fifth aspect of the present invention provides a vector, which contains the nucleic acid molecule described in the fourth aspect of the present invention.
- the sixth aspect of the present invention provides a host cell containing the vector of the fifth aspect of the present invention.
- a seventh aspect of the present invention provides a molecule capable of binding the peptide of the first aspect of the present invention and/or the pMHC complex of the second aspect of the present invention.
- the molecule can specifically bind to the peptide described in the first aspect of the present invention and/or the pMHC complex described in the second aspect of the present invention.
- the molecule is a T cell receptor.
- the T cell receptor is soluble.
- the molecule is an antibody or a binding fragment thereof.
- the antibody is a monoclonal antibody.
- an isolated monoclonal T cell obtained by utilizing the peptide of the first aspect of the present invention and/or the pMHC complex of the second aspect of the present invention and/or the present invention
- the cells are obtained by separation.
- the monoclonal T cells specifically bind to the pMHC complex of the second aspect of the present invention.
- the ninth aspect of the present invention provides the use of the peptide of the first aspect of the present invention, the pMHC complex of the second aspect of the present invention or the cell of the third aspect of the present invention for activating and/or isolating T cells.
- the tenth aspect of the present invention provides the use of the peptide of the first aspect of the present invention and the pMHC complex of the second aspect of the present invention for screening T cell receptors or antibody libraries.
- the eleventh aspect of the present invention provides the peptide of the first aspect of the present invention, the pMHC complex of the second aspect of the present invention, the cell of the third aspect of the present invention, the nucleic acid molecule of the fourth aspect of the present invention,
- the use of the molecule described in the seventh aspect of the present invention or the T cell described in the eighth aspect of the present invention is for preparing a medicament for preventing or treating cancer.
- the twelfth aspect of the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier, the peptide of the first aspect of the present invention, the pMHC complex of the second aspect of the present invention, the The cell of the third aspect, the molecule of the seventh aspect of the present invention, or the T cell of the eighth aspect of the present invention.
- the pharmaceutical composition is a vaccine.
- the thirteenth aspect of the present invention provides a method for preventing or treating diseases, comprising administering an appropriate amount of the peptide of the first aspect of the present invention, the pMHC complex of the second aspect of the present invention, the first aspect of the present invention to a subject in need.
- the fourteenth aspect of the present invention provides a method for obtaining a molecule that binds to the pMHC complex described in the second aspect of the present invention, comprising:
- Figure 1 is a representative mass spectrum for the identification of the short peptides of the present invention.
- Figure 2 is a native gel map of the soluble pMHC complex of the present invention. Left band is BSA control, right band is pMHC complex.
- Figure 3 shows the double positive staining results of CD8+ and tetramer-PE of T cell clones.
- Fig. 4 is a graph showing the results of the Elispot function experiment of the monoclonal cells obtained by using the short peptide of the present invention on tumor cell lines and T2 cells.
- Figure 5 is a kinetic map of the binding of soluble TCR molecules obtained by using the short peptides of the present invention to the pMHC complexes of the present invention.
- the present invention obtains a peptide derived from the antigen SSX2, which is presented on the surface of tumor cells by MHC molecules as a tumor marker. Accordingly, the present invention provides peptides derived from the antigen SSX2, complexes formed by such peptides with MHC molecules and uses of said peptides and complexes. At the same time, the present invention also relates to molecules that bind to the above-mentioned peptides or complexes.
- the peptide of the present invention and the polypeptide of the present invention or the short peptide of the present invention can be used interchangeably, and both refer to the peptide derived from the antigen SSX2 provided by the present invention.
- the first aspect of the present invention provides a peptide, the amino acid sequence of the peptide is: AQIPEKIQK (SEQ ID NO: 1).
- the peptides of the present invention may be post-translationally modified at one or more positions between the amino acid sequences.
- post-translational modifications can be found in Engelhard et al. Curr Opin Immunol. 2006 Feb;18(1):92-7, and include phosphorylation, acetylation, and deamidation.
- the peptide of the present invention binds to MHC at the peptide binding site of the MHC molecule.
- the modified amino acids described above do not disrupt the ability of the peptide to bind to MHC.
- the amino acid modification increases the ability of the peptide to bind to MHC.
- mutations may occur at the binding site of the peptide to the MHC.
- the peptides of the present invention may be composed of AQIPEKIQK (SEQ ID NO: 1), or mainly composed of AQIPEKIQK (SEQ ID NO: 1), which correspond to the positions of amino acid residues 15-23 of the full length of the SSX2 protein.
- the invention also provides analogs of the protein or peptide shown in SEQ ID NO: 1. Differences between these analogs and natural peptides may be differences in amino acid sequence, differences in modified forms that do not affect the sequence, or both. These peptides include natural or induced genetic variants. Induced variants can be obtained by a variety of techniques, such as random mutagenesis by radiation or exposure to mutagens, but also by site-directed mutagenesis or other known molecular biology techniques. Analogs also include analogs with residues other than natural L-amino acids (eg, D-amino acids), as well as analogs with non-naturally occurring or synthetic amino acids (eg, beta, gamma-amino acids). It should be understood that the peptides of the present invention are not limited to the representative peptides exemplified above.
- Modified (usually without altering the primary structure) forms include chemically derivatized forms of the peptide, such as acetylation or carboxylation, in vivo or in vitro. Modifications also include glycosylation, such as those resulting from glycosylation modifications in peptide synthesis and processing or in further processing steps. This modification can be accomplished by exposing the peptide to enzymes that perform glycosylation, such as mammalian glycosylases or deglycosylases. Modified forms also include sequences with phosphorylated amino acid residues (eg, phosphotyrosine, phosphoserine, phosphothreonine). Also included are peptides modified to increase their resistance to proteolysis or to optimize solubility.
- the peptides of the present invention can be synthesized simply by the Merrifield synthesis method (also known as polypeptide solid-phase synthesis). GMP grade peptides can be synthesized using solid phase synthesis techniques at Multiple Peptide Systems (San Diego, CA). Alternatively, the peptides can be synthesized recombinantly, if desired, by methods known in the art. Typical of such methods involve the use of vectors comprising nucleic acid sequences encoding the polypeptides to express the polypeptides in vivo; eg, in bacterial, yeast, insect or mammalian cells. Alternatively, in vitro cell-free systems can also be used for expression. Such systems are known in the art and are commercially available.
- the peptides may be isolated and/or provided in substantially pure form. For example, they may be provided in a form substantially free of other peptides or proteins.
- a second aspect of the present invention provides a pMHC complex comprising the peptide of the first aspect of the present invention.
- the polypeptide is bound to the peptide-binding groove of the MHC molecule.
- the MHC molecule may be an MHC class I molecule or an MHC class II molecule, preferably, the MHC molecule is an MHC class I molecule.
- the MHC molecule is HLA-A*11, more preferably, the MHC molecule is HLA-A*1101.
- the pMHC complexes of the present invention may exist in multimeric form, eg, dimers, or tetramers, or pentamers, or hexamers, or octamers, or larger. Appropriate methods for generating pMHC multimers can be found in the relevant literature, eg (Greten et al., Clin. Diagnostic Lab. Immunol. 2002:216-220).
- pMHC multimers can be produced by complexing pMHC complexes with biotin residues in combination with fluorescently labeled streptavidin.
- the pMHC multimers can also be formed by immunoglobulins as molecular scaffolds. In this system, the extracellular region of the MHC molecule is joined to the constant region of the immunoglobulin heavy chain by a short linker.
- the formation of pMHC multimers may utilize carrier molecules such as dextran (WO02072631). pMHC multimers help improve detection of moieties bound to them, such as T cell receptors. Alternatively, enhance the effect of pMHC complexes in related applications, such as activation of T cells.
- the pMHC complexes of the present invention may be provided in soluble form.
- the MHC molecules in the pMHC complexes do not contain a transmembrane region.
- an MHC class I molecule may consist of the extracellular domain of its light chain and all or part of its heavy chain.
- the MHC molecule is a fragment comprising only its functional domain.
- MHC molecules in the soluble pMHC complexes of the invention can also be produced synthetically and then refolded with the peptides of the invention. By determining whether the peptides and MHC molecules are capable of refolding, it is possible to determine which class of MHC molecules the peptides of the invention are capable of forming complexes with.
- the soluble pMHC complexes of the present invention can be used to screen or detect molecules, such as TCRs or antibodies, to which they bind.
- the method includes contacting the pMHC complex with a binding moiety to be tested, and determining whether the binding moiety to be tested is bound to the complex.
- Methods for assaying binding of pMHC complexes are well known in the art. Preferred methods include, but are not limited to, surface plasmon resonance, or any other biosensing technique, ELISA, flow cytometry, chromatography, microscopy.
- the binding can be detected by functional assays of the biological response to binding, such as cytokine release or apoptosis.
- the soluble pMHC complexes of the invention can also be used to screen TCR or antibody libraries.
- the construction of antibody libraries using phage display technology is well known in the art, as described in reference Aitken, Antibody phage display: Methods and Protocols (2009, Humana, New York).
- the pMHC complexes of the invention are used to screen diverse TCR libraries displayed on the surface of phage particles.
- the TCRs displayed by the library may contain non-native mutations.
- the soluble pMHC complexes of the present invention can be immobilized on a suitable solid support via a linker.
- suitable solid supports include, but are not limited to, beads, membranes, agarose gels, magnetic beads, substrates, tubes, columns.
- the pMHC complexes can be immobilized on ELISA reaction plates, magnetic beads, or surface plasmon resonance biosensor chips.
- Methods of immobilizing pMHC complexes to solid supports are known to those skilled in the art and include, for example, the use of affinity binding pairs such as biotin and streptavidin, or antibodies and antigens.
- the pMHC complex is labeled with biotin and immobilized on a streptavidin-coated surface.
- the peptides of the present invention can be presented to the cell surface together with MHC complexes. Accordingly, the present invention also provides a cell capable of presenting the pMHC complexes of the present invention to its surface.
- Such cells may be mammalian cells, preferably immune system cells, and preferably specialized antigen presenting cells, such as dendritic cells or B cells.
- Other preferred cells include T2 cells (Hosken, et al., Science. 1990. 248:367-70).
- Cells presenting the peptides or pMHC complexes of the present invention may be isolated, preferably, in the form of a population of cells, or provided in substantially pure form.
- the cells may not naturally present the complexes of the invention, or the cells may present higher levels of the complexes than in the native state.
- Such cells can be obtained by pulsing with the peptides of the present invention. Pulse treatment involves incubating cells with the peptide for several hours, preferably at a concentration of 10-5-10-12M .
- the cells can also be transduced with HLA-A*11 molecules to further induce peptide presentation.
- Cells presenting the pMHC complexes of the present invention can be used to isolate T cells and T cell receptors, which are activated by said cells and further sorted, and can also obtain expression in said T cells. surface T cell receptors.
- the method of obtaining the above-mentioned T cells comprises stimulating fresh blood obtained from healthy volunteers with the above-mentioned cells presenting the pMHC complexes of the present invention. You can go through several rounds of stimulation, such as 3-4 rounds. Identification of activated T cells can be determined by measuring cytokine release in the presence of peptide-pulsed T2 cells of the invention (eg, IFN- ⁇ ELISpot assay). Using labeled antibodies, activated cells can be sorted by flow cytometry (FACS), and sorted cells can be expanded in culture and further validated, for example, by ELISpot detection and/or cytotoxicity against target cells and/or pMHC multimerization Body staining was verified. TCR chains from validated T cell clones can be amplified by rapid amplification of cDNA ends (RACE) and sequenced.
- RACE rapid amplification of cDNA ends
- the present invention also provides a nucleic acid molecule comprising a nucleic acid sequence encoding the peptide of the present invention.
- the nucleic acid may be cDNA.
- the nucleic acid molecule may consist essentially of nucleic acid sequences encoding the peptides of the invention, or may only encode the peptides of the invention.
- Such nucleic acid molecules can be synthesized using methods known in the art. Due to the degeneracy of the genetic code, those skilled in the art will understand that nucleic acid molecules of different nucleic acid sequences may encode the same amino acid sequence.
- the present invention also provides a vector, which includes the nucleic acid sequence of the present invention.
- Suitable vectors are known in the art of vector construction, including promoter selection and other regulatory elements, such as enhancer elements.
- the vectors of the present invention include sequences suitable for introduction into cells.
- the vector can be an expression vector, in which the coding sequence of the polypeptide is controlled by its own cis-acting regulatory elements, and the design of the vector is convenient for gene integration or gene replacement in host cells.
- vector includes DNA molecules, such as plasmids, phages, viruses or other vectors, which contain one or more heterologous or recombinant nucleic acid sequences.
- Suitable phage and viral vectors include, but are not limited to: lambda-phage, EMBL phage, simian virus, bovine wart virus, Epstein-Barr virus, adenovirus, herpes virus, mouse sarcoma virus, murine breast cancer virus, lentivirus, etc. .
- the present invention also provides a binding molecule that can be used as an immunotherapeutic or diagnostic agent.
- the binding molecule can bind only to the peptide, or to a complex formed by the peptide and the MHC molecule. In the latter case, the binding molecule may be partially bound to the MHC molecule, while at the same time it is also bound to the peptide of the invention.
- the binding moieties of the present invention may be isolated and/or soluble, and/or non-naturally occurring, ie, with no equivalents found in nature, and/or pure, and/or artificially synthesized.
- the binding molecule is a T cell receptor (TCR).
- TCRs can be described using the International Information System for Immunogenetics (IMGT).
- IMGT International Information System for Immunogenetics
- Native ⁇ heterodimeric TCRs have ⁇ and ⁇ chains. Broadly speaking, each chain contains a variable region, a linker region and a constant region, and the beta chain typically also contains a short variable region between the variable region and the linker region, but the variable region is often considered part of the linker region.
- the TCR of the present invention may be in any form known in the art.
- the TCR may be a heterodimer, or exist as a single chain.
- the TCR may be in a soluble form (ie no transmembrane or cytoplasmic domain), in particular, the TCR may comprise all or part of the TCR extracellular domain.
- the TCR may also be a full-length chain comprising its transmembrane region.
- the TCR can be presented to the surface of cells, such as T cells.
- Soluble TCRs can be obtained by combining existing techniques in the art, for example, by introducing artificial disulfide bonds between the constant domains of the ⁇ and ⁇ chains of ⁇ TCR, or between the ⁇ chain variable region and the ⁇ chain constant region of ⁇ TCR. artificial disulfide bonds were introduced.
- the TCRs of the invention can be used to deliver cytotoxic or immunostimulatory agents to target cells, or be transformed into T cells, enabling T cells expressing the TCR to destroy tumor cells for use in a treatment process known as adoptive immunotherapy given to the patient.
- the TCR of the present invention may also contain mutations, and preferably, the affinity of the mutated TCR to the pMHC complex of the present invention is improved.
- the TCR of the present invention can be used alone, or can be combined with the conjugate in a covalent or other manner, preferably in a covalent manner.
- the conjugate includes a detectable label (for diagnostic purposes, wherein the TCR is used to detect the presence of cells presenting the pMHC complexes of the invention), a therapeutic agent, a PK (protein kinase) modification moiety, or a combination of any of the above. Combination binding or conjugation.
- the TCRs of the present invention can also bind to anti-CD3 antibodies, preferably covalently, to redirect T cells to kill target cells.
- the binding molecule of the present invention is an antibody.
- antibody refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, ie, molecules containing specific binding sites, which may be all-natural, partially synthetic, or fully synthetic.
- antibody includes antibody fragments, derivatives thereof, functional equivalents thereof, and homologous antibodies, humanized antibodies, which antibody fragments include an immunoglobulin binding region that is or binds to an antibody Region homology. It can be all natural, or partially synthetic, or totally synthetic.
- a humanized antibody can be a modified antibody that contains the variable regions of a non-human antibody (eg, mouse) and the constant regions of a human antibody.
- antibodies may be isotype immunoglobulins (eg, IgG, IgE, IgM, IgD, and IgA) and subclasses of their isotypes; fragments include antigen binding regions, such as Fab, scFv, Fv, dAb, Fd; and diabodies.
- Antibodies can be polyclonal or monoclonal, preferably monoclonal.
- TCR and antibody preparation methods of the above-mentioned TCR and antibody are known to those skilled in the art, including but not limited to, expression from E. coli cells or insect cells, and purification.
- the present invention further provides the use of the peptides, pMHC complexes, nucleic acid molecules, vectors, cells and binding molecules of the present invention in pharmaceuticals.
- the peptides, pMHC complexes, nucleic acids, vectors, cells or binding molecules can be used to treat or prevent cancer, preferably melanoma, bladder cancer, liver cancer, epidermoid cancer, non-small cell lung cancer, squamous cell cancer, and the like.
- the present invention also provides a pharmaceutical composition
- a pharmaceutical composition comprising the peptide of the present invention, the pMHC complex, the nucleic acid molecule of the present invention, the cell of the present invention, or the binding molecule of the present invention, and a pharmaceutically acceptable carrier.
- the pharmaceutical composition may be in any suitable form, (depending on the method of administration desired by the patient). It can be presented in unit dosage form, usually in a sealed container, and can be presented as part of a kit. Such kits usually, but not necessarily, contain instructions for use. It may contain a plurality of said unit dosage forms.
- compositions are suitable for any suitable route of administration, such as injection (including subcutaneous, intramuscular, intraperitoneal or intravenous), inhalation or oral, or nasal, or anal routes.
- routes of administration such as injection (including subcutaneous, intramuscular, intraperitoneal or intravenous), inhalation or oral, or nasal, or anal routes.
- the compositions may be prepared by any method known in the art of pharmacy, eg, by admixing the active ingredient with a carrier or excipient under sterile conditions.
- the dosage administered of the formulations of the present invention may vary widely.
- the appropriate dose to be administered will be ultimately determined by the physician.
- peptides, pMHC complexes, or cells presenting pMHC complexes, which are presented to the cell surface along with MHC molecules can activate T cells or B cells to function.
- the peptides, pMHC complexes or cells presenting the pMHC complexes of the invention may be provided in the form of vaccine compositions.
- the vaccine composition can be used to treat or prevent cancer. All such compositions are included in the present invention. It will be appreciated that the vaccine may be in a variety of forms (Schlom J.J Natl Cancer Inst. 2012 104(8):599-613).
- the peptides of the invention can be used directly to immunize patients (Salgaller ML. Cancer Res. 1996.56(20):4749-57 and Marchand M.Int J Cancer. 1999.80(2):219-230).
- the vaccine composition may contain additional peptides such that the peptide of the invention is one of a mixture of peptides.
- the vaccine composition may be adjuvanted to enhance the immune response.
- the vaccine composition may be in the form of an antigen presenting cell presenting the peptide of the invention and the MHC complex.
- the antigen presenting cells are immune cells, more preferably dendritic cells.
- the peptides can also be pulsed onto the surface of cells (Thurner BI. et al., J. Exp. Med. 1999. 190:1669), or nucleic acids encoding the peptides of the invention can be introduced into dendritic cells, eg, by electroporation Law (Van Tendeloo, VF. et al., Blood 2001.98:49).
- the present invention uses a digital single-molecule multiplex gene expression profiling system for detection (nanostring), which further verifies the large expression of SSX2 antigen in liver cancer cells.
- the HLA-short peptide complexes were purified using the commercial antibody A11.1M. Specifically, tumor cells were lysed with a buffer containing a non-ionic surfactant Triton X-100 (1% v/v), 1 ml of lysis buffer was added to 2*10 ⁇ 7 cells, and the cells were incubated at 4°C with rolling for 1 h. Cell debris was removed by centrifugation, the supernatant was incubated with the antibody, and then rProtein A-Sepharose was added to capture the "antibody-HLA-short peptide complex". Pass the column to collect "rProtein A-Sepharose-antibody-HLA-short peptide complex".
- the column was washed with low-salt and high-salt buffers, and finally the HLA-short peptide complexes on the immunoaffinity column were eluted with 10% acetic acid, heated at 95°C, and subjected to 10KDa (AmiconR Ultr Centrifugal Filters, MILLIPORE) supernatant.
- 10KDa AmiconR Ultr Centrifugal Filters, MILLIPORE
- the peptide mixture was fractionated by Agilent 1260 high performance liquid chromatography: ZORBAX 300SB-C18; 1.0*150mm, 3.5um; mobile phase A was 98% water, 2% acetonitrile, 0.1% trifluoroacetic acid, mobile phase B was 98% acetonitrile, 2 % water, 0.1% trifluoroacetic acid, mobile phase gradient from 5% to 70% mobile phase B over 10 minutes. One fraction was collected every minute. The total run time is 30 minutes.
- HPLC fractions of the peptides were concentrated and injected into the nanoLC-MSMS system for analysis:
- Eksigent nanoLC-AB Sciex Triple TOF 5600 system Mass spectrometry using IDA analysis method. Liquid chromatography adopts: pre-column: (Eksigent) NanoLC Trap column.5 ⁇ m C18.100 ⁇ m*2.5cm, 910-00050, analytical column: (Eksigent) C18-CL-120, 3 ⁇ m, 0.075 ⁇ 150mm, 805-00120.
- Dionex Ultimate3000-Thermo QE Plus system Mass spectrometry adopts ddms2 analysis method.
- Liquid chromatography adopts: Pre-column: (Thermo) Acclaim 100um ⁇ 2cm, nanoViper, C18, 5um, 100A, 164564, analytical column: (Thermo) Acclaim 75um ⁇ 15cm, nanoViper, C18, 3um, 100A, 164568.
- the mobile phase A of the nanoflow chromatography of the above two systems was 98% water, 2% acetonitrile, 0.1% formic acid, and the mobile phase B was 98% acetonitrile, 2% water, 0.1% formic acid, and the mobile phase gradient was mobile phase within 74 minutes. B increased from 5% to 50%. The total run time is 90 minutes.
- the heavy chain and light chain ( ⁇ 2m) of type I HLA-A*1101 molecules were expressed in E. coli in the form of inclusion bodies, respectively. It should be noted that in order to obtain soluble pMHC complexes, the heavy chain of the HLA-A*1101 molecule used in this example does not contain its transmembrane and cytoplasmic regions. In addition, to facilitate subsequent biotinylation of the soluble pMHC complex, a biotinylation tag can be added to the C-terminus of the heavy chain.
- the specific process of preparing the soluble pMHC complex of the present invention is as follows:
- Collect 100ml of E.coli bacteria that induces the expression of heavy or light chains centrifuge at 8000g at 4°C for 10min, wash the cells once with 10ml PBS, and then use 5ml BugBuster Master Mix Extraction Reagents (Merck) to vigorously shake the cells to resuspend the cells. Incubate with rotation at room temperature for 20 min, then centrifuge at 6000g for 15 min at 4°C, discard the supernatant, and collect the inclusion bodies.
- the peptide AQIPEKIQK of the present invention was dissolved in DMSO to a concentration of 20 mg/ml.
- the inclusion bodies of light chain and heavy chain were dissolved with 8M urea, 20mM Tris pH 8.0, 10mM DTT, and further denatured by adding 3M guanidine hydrochloride, 10mM sodium acetate, 10mM EDTA before renaturation.
- AQIPEKIQK peptide was added to renaturation buffer (0.4M L-arginine, 100mM Tris pH 8.3, 2mM EDTA, 0.5mM oxidized glutathione, 5mM reduced glutathione, 0.2mM PMSF, cooled to 4°C), then added 20mg/L light chain and 90mg/L heavy chain in sequence (final concentration, heavy chain was added in three times, 8h/time), and renatured at 4°C for at least 3 days to completion.
- renaturation buffer 0.4M L-arginine, 100mM Tris pH 8.3, 2mM EDTA, 0.5mM oxidized glutathione, 5mM reduced glutathione, 0.2mM PMSF, cooled to 4°C
- renaturation buffer by dialyzing against 10 volumes of 20 mM Tris pH 8.0, at least twice, to sufficiently reduce the ionic strength of the solution.
- the protein solution was filtered through a 0.45 ⁇ m cellulose acetate filter and loaded onto a HiTrap Q HP (GE) anion exchange column (5 ml bed volume).
- the protein was eluted using an Akta purifier (GE), a linear gradient of 0-400 mM NaCl prepared in 20 mM Tris pH 8.0, and pMHC was eluted at approximately 250 mM NaCl, and peak fractions were collected.
- the native gel map of the obtained soluble pMHC complex of the present invention is shown in Figure 2, and the bands are very uniform.
- Purified pMHC molecules were concentrated with Millipore ultrafiltration tubes while buffer exchanged to 20mM Tris pH 8.0, followed by addition of biotinylation reagents 0.05M Bicine pH 8.3, 10mM ATP, 10mM MgOAc, 50 ⁇ M D-Biotin, 100 ⁇ g/ml BirA Enzyme (GST-BirA), the mixture was incubated overnight at room temperature, and the complete biotinylation was checked by SDS-PAGE.
- the biotinylated pMHC molecules were concentrated to 1 ml with a Millipore ultrafiltration tube, and the biotinylated pMHC was purified by gel filtration chromatography. HiPrepTM was pre-equilibrated with filtered PBS using an Akta purifier (GE). A 16/60S200HR column (GE), loaded with 1 ml of concentrated biotinylated pMHC molecules, was then eluted with PBS at a flow rate of 1 ml/min.
- Akta purifier Akta purifier
- This example provides an illustration of the use of the pMHC complexes of the invention to obtain monoclonal T cells.
- TCR sequences can be cloned into a suitable vector and then expressed in E. coli, such as E. coli, or on the surface of phage.
- peripheral blood lymphocytes of healthy volunteers are stimulated with the short peptide of the present invention, sorted, and then monoclonally cultured by the limiting dilution method to obtain T cell clones.
- the CD8+ and tetramer-PE double positive staining results are shown in Figure 3 Show.
- the function and specificity of the T cell clone were further tested by ELISPOT assay.
- the specific response of the T cell clones obtained by using the short peptide of the present invention to the tumor cell line can also indicate that the short peptide of the present invention is indeed presented to the surface of tumor cells by MHC.
- the effector cells used in the IFN- ⁇ ELISPOT experiment in this example are the T cell clones obtained in the present invention
- the target cells are T2 cells loaded with the short peptides of the present invention (transformed into HLA A1101 into T2), and T2 cells loaded with other short peptides Cells (transfected with HLA A1101 into T2), negative tumor cell line K562 and positive tumor cell line K562 (A11, SSX2-P2A-GFP) (transfected with HLA A1101 and SSX2 antigen and GFP), which are loaded with other short peptides T2 cells and tumor cell line K562 served as controls.
- the ELISPOT experiment steps are as follows: Add the components of the test to the ELISPOT plate in the following order: 20,000 target cells/well, 2000 effector cells/well, the amount of short peptide added in the T2 experimental group is 20 ⁇ l, tumor cells 20 ⁇ l culture medium (test medium) was added to the line group, and 2 duplicate wells were set up. It was then incubated overnight (37°C, 5% CO2 ). The plates were then washed and subjected to secondary detection and color development, the plates were dried for 1 hour, and the spots formed on the membrane were counted using an immunospot plate reader (ELISPOT READER system; AID Corporation).
- ELISPOT READER system AID Corporation
- the experimental results are shown in Fig. 4.
- the obtained specific antigen-specific T cell clones have specific responses to T2 cells loaded with the short peptide of the present invention, but basically no response to T2 cells loaded with other irrelevant peptides.
- T cell clones obtained using the short peptides of the present invention have specific responses to positive tumor cell lines, but no response to negative tumor cell lines. It shows that functional T cell clones are obtained by the short peptides of the present invention.
- RNA of the above T cell clones was extracted with Quick-RNA TM MiniPrep (ZYMO research), and the TCR sequence was obtained.
- the soluble TCR protein was expressed in E. coli in this example, and its binding to the pMHC complex was detected by BIAcore. It should be noted that soluble TCRs can be obtained according to the prior art, including but not limited to, those described in patent document PCT/CN2015/093806.
- the BIAcore T200 real-time analysis system was used to detect the binding activity of soluble TCR protein to pMHC complexes.
- Anti-streptavidin antibody (GenScript) was added to coupling buffer (10 mM sodium acetate buffer, pH 4.77), and the antibody was then flowed through a CM5 chip preactivated with EDC and NHS to immobilize the antibody on the chip. surface, and finally blocked the unreacted activated surface with ethanolamine hydrochloric acid solution to complete the coupling process with a coupling level of about 15,000 RU.
- a low concentration of streptavidin was flowed over the antibody-coated chip surface, then pMHC prepared in the manner described in Example 2 was flowed through the detection channel, the other channel was used as a reference channel, and 0.05 mM The biotin was flowed through the chip at a flow rate of 10 ⁇ L/min for 2 min to block the remaining binding sites of streptavidin.
- Figure 5 shows the kinetic map of the combination of soluble TCR molecules and pMHC complexes obtained by using the short peptide of the present invention, and the map shows the combination of the two.
- the above method was used to detect the binding activity of the soluble TCR molecule to several other irrelevant antigen short peptides and HLA complexes, and the results showed that the TCR molecule of the present invention did not bind to other irrelevant antigens.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Wood Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Urology & Nephrology (AREA)
- General Engineering & Computer Science (AREA)
- Oncology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- General Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
Abstract
La présente invention concerne des peptides courts dérivés de l'antigène SSX2, et en particulier des peptides courts d'antigène tumoral dérivés de SSX2, des complexes formés par les peptides courts et les molécules MHC, et des utilisations des peptides courts et des complexes. De plus, la présente invention concerne également des molécules liées aux peptides ou complexes courts décrits ci-dessus, et des utilisations des molécules.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011324186.8A CN114524870A (zh) | 2020-11-23 | 2020-11-23 | 源自于ssx2抗原的短肽 |
CN202011324186.8 | 2020-11-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2022105922A1 true WO2022105922A1 (fr) | 2022-05-27 |
Family
ID=81618791
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2021/132192 WO2022105922A1 (fr) | 2020-11-23 | 2021-11-22 | Peptides courts dérivés d'un antigène ssx2 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN114524870A (fr) |
WO (1) | WO2022105922A1 (fr) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117304301A (zh) * | 2022-06-17 | 2023-12-29 | 香雪生命科学技术(广东)有限公司 | 一种结合ssx2抗原的tcr分子及其应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1691964A (zh) * | 2002-09-06 | 2005-11-02 | 曼康公司 | 表位序列 |
CN101287755A (zh) * | 2005-09-05 | 2008-10-15 | 伊玛提克斯生物技术有限公司 | 结合于人类白细胞抗原(hla)ⅰ类或ⅱ类分子的肿瘤相关肽及相关的抗癌疫苗 |
CN110343167A (zh) * | 2018-04-03 | 2019-10-18 | 广东香雪精准医疗技术有限公司 | 识别ssx2抗原短肽的t细胞受体 |
CA3100775A1 (fr) * | 2018-05-18 | 2019-11-21 | Children's National Medical Center | Therapie ciblee amelioree par lymphocytes t |
CN110950949A (zh) * | 2018-09-26 | 2020-04-03 | 广东香雪精准医疗技术有限公司 | 一种识别ssx2抗原的t细胞受体 |
-
2020
- 2020-11-23 CN CN202011324186.8A patent/CN114524870A/zh active Pending
-
2021
- 2021-11-22 WO PCT/CN2021/132192 patent/WO2022105922A1/fr unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1691964A (zh) * | 2002-09-06 | 2005-11-02 | 曼康公司 | 表位序列 |
CN101287755A (zh) * | 2005-09-05 | 2008-10-15 | 伊玛提克斯生物技术有限公司 | 结合于人类白细胞抗原(hla)ⅰ类或ⅱ类分子的肿瘤相关肽及相关的抗癌疫苗 |
CN110343167A (zh) * | 2018-04-03 | 2019-10-18 | 广东香雪精准医疗技术有限公司 | 识别ssx2抗原短肽的t细胞受体 |
CA3100775A1 (fr) * | 2018-05-18 | 2019-11-21 | Children's National Medical Center | Therapie ciblee amelioree par lymphocytes t |
CN110950949A (zh) * | 2018-09-26 | 2020-04-03 | 广东香雪精准医疗技术有限公司 | 一种识别ssx2抗原的t细胞受体 |
Also Published As
Publication number | Publication date |
---|---|
CN114524870A (zh) | 2022-05-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20210061874A1 (en) | Claudin-6 peptides | |
US20210121547A1 (en) | Progesterone-associated endometrial protein (paep) and uses thereof | |
CN111138521B (zh) | 源自于afp抗原的短肽 | |
US20230190901A1 (en) | Peptides derived from melanoma-associated antigen c2 (magec2) and uses thereof | |
US20210322528A1 (en) | Peptides from npsr1 | |
US20210122798A1 (en) | Peptides derived from achaete-scute homolog 2 (ascl2), complexes comprising such peptides bound to mhc molecules | |
CN107936109B (zh) | 衍生自sage1的肿瘤抗原短肽 | |
US20180334476A1 (en) | Peptides | |
WO2022105922A1 (fr) | Peptides courts dérivés d'un antigène ssx2 | |
US20210139558A1 (en) | Peptides | |
US20210170003A1 (en) | Peptides derived from homeobox protein b13 (hox-b13) and complexes comprising such peptides bound to mhc molecules | |
US20210145936A1 (en) | Peptides derived from actin-like protein 8 (actl8) | |
CN108218977B (zh) | 源自于肿瘤抗原sage1的短肽 | |
CN108250289B (zh) | 源自于mage b6的肿瘤抗原短肽 | |
CN112300261B (zh) | 一种衍生自afp的肿瘤抗原短肽 | |
CN111138522B (zh) | 衍生自afp的肿瘤抗原短肽 | |
CN110862449B (zh) | 衍生自ssx的肿瘤抗原短肽 | |
CN108250286B (zh) | 源自于pasd1的肿瘤抗原短肽 | |
CN108203460B (zh) | 衍生自肿瘤抗原prame的短肽 | |
CN107266552B (zh) | 源自于prame的肿瘤抗原短肽 | |
CN108997481B (zh) | 源自于lmp1的抗原短肽 | |
CN108218976B (zh) | 衍生自lmp1的肿瘤抗原短肽 | |
CN112898399A (zh) | 源自于afp抗原的短肽 | |
CN116948001A (zh) | 一种抗原短肽 | |
CN116948002A (zh) | 一种肽及其复合物 |