CN114276448A - 抗dm1单克隆抗体及其制备方法与应用 - Google Patents
抗dm1单克隆抗体及其制备方法与应用 Download PDFInfo
- Publication number
- CN114276448A CN114276448A CN202111577267.3A CN202111577267A CN114276448A CN 114276448 A CN114276448 A CN 114276448A CN 202111577267 A CN202111577267 A CN 202111577267A CN 114276448 A CN114276448 A CN 114276448A
- Authority
- CN
- China
- Prior art keywords
- monoclonal antibody
- seq
- antibody
- variable region
- gly
- 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.)
- Withdrawn
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 239000000427 antigen Substances 0.000 claims abstract description 14
- 102000036639 antigens Human genes 0.000 claims abstract description 14
- 108091007433 antigens Proteins 0.000 claims abstract description 14
- 229940079593 drug Drugs 0.000 claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 10
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims abstract description 8
- 239000003446 ligand Substances 0.000 claims abstract description 8
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 5
- 150000007523 nucleic acids Chemical class 0.000 claims description 8
- 102000039446 nucleic acids Human genes 0.000 claims description 7
- 108020004707 nucleic acids Proteins 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 230000008878 coupling Effects 0.000 abstract description 6
- 238000010168 coupling process Methods 0.000 abstract description 6
- 238000005859 coupling reaction Methods 0.000 abstract description 6
- 210000004027 cell Anatomy 0.000 description 14
- 239000000243 solution Substances 0.000 description 11
- 150000001413 amino acids Chemical group 0.000 description 10
- 241001529936 Murinae Species 0.000 description 9
- 206010003445 Ascites Diseases 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 238000002649 immunization Methods 0.000 description 7
- 230000003053 immunization Effects 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 5
- ANZJBCHSOXCCRQ-FKUXLPTCSA-N mertansine Chemical compound CO[C@@H]([C@@]1(O)C[C@H](OC(=O)N1)[C@@H](C)[C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(=O)CCS)CC(=O)N1C)\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 ANZJBCHSOXCCRQ-FKUXLPTCSA-N 0.000 description 5
- 229960005558 mertansine Drugs 0.000 description 5
- 102000014914 Carrier Proteins Human genes 0.000 description 4
- 108010078791 Carrier Proteins Proteins 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- 239000000611 antibody drug conjugate Substances 0.000 description 3
- 229940049595 antibody-drug conjugate Drugs 0.000 description 3
- 239000000562 conjugate Substances 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- -1 small molecule compounds Chemical class 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- QDRGPQWIVZNJQD-CIUDSAMLSA-N Ala-Arg-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O QDRGPQWIVZNJQD-CIUDSAMLSA-N 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- WNZOCXUOGVYYBJ-CDMKHQONSA-N Gly-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)CN)O WNZOCXUOGVYYBJ-CDMKHQONSA-N 0.000 description 2
- HQSKKSLNLSTONK-JTQLQIEISA-N Gly-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 HQSKKSLNLSTONK-JTQLQIEISA-N 0.000 description 2
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 2
- ZNTSGDNUITWTRA-WDSOQIARSA-N His-Trp-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C(C)C)C(O)=O ZNTSGDNUITWTRA-WDSOQIARSA-N 0.000 description 2
- NZOCIWKZUVUNDW-ZKWXMUAHSA-N Ile-Gly-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O NZOCIWKZUVUNDW-ZKWXMUAHSA-N 0.000 description 2
- QJXHMYMRGDOHRU-NHCYSSNCSA-N Leu-Ile-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O QJXHMYMRGDOHRU-NHCYSSNCSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- DJPXNKUDJKGQEE-BZSNNMDCSA-N Phe-Asp-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DJPXNKUDJKGQEE-BZSNNMDCSA-N 0.000 description 2
- MCIXMYKSPQUMJG-SRVKXCTJSA-N Phe-Ser-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MCIXMYKSPQUMJG-SRVKXCTJSA-N 0.000 description 2
- YMTLKLXDFCSCNX-BYPYZUCNSA-N Ser-Gly-Gly Chemical compound OC[C@H](N)C(=O)NCC(=O)NCC(O)=O YMTLKLXDFCSCNX-BYPYZUCNSA-N 0.000 description 2
- XKFJENWJGHMDLI-QWRGUYRKSA-N Ser-Phe-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O XKFJENWJGHMDLI-QWRGUYRKSA-N 0.000 description 2
- JWHOIHCOHMZSAR-QWRGUYRKSA-N Tyr-Asp-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 JWHOIHCOHMZSAR-QWRGUYRKSA-N 0.000 description 2
- JAIZPWVHPQRYOU-ZJDVBMNYSA-N Val-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C(C)C)N)O JAIZPWVHPQRYOU-ZJDVBMNYSA-N 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 108020001507 fusion proteins Proteins 0.000 description 2
- 102000037865 fusion proteins Human genes 0.000 description 2
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 2
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 2
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 2
- 108010089804 glycyl-threonine Proteins 0.000 description 2
- 108010045126 glycyl-tyrosyl-glycine Proteins 0.000 description 2
- 108010081551 glycylphenylalanine Proteins 0.000 description 2
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- UZOVYGYOLBIAJR-UHFFFAOYSA-N 4-isocyanato-4'-methyldiphenylmethane Chemical compound C1=CC(C)=CC=C1CC1=CC=C(N=C=O)C=C1 UZOVYGYOLBIAJR-UHFFFAOYSA-N 0.000 description 1
- 206010000060 Abdominal distension Diseases 0.000 description 1
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 1
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- UWIQWPWWZUHBAO-ZLIFDBKOSA-N Ala-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)CC(C)C)C(O)=O)=CNC2=C1 UWIQWPWWZUHBAO-ZLIFDBKOSA-N 0.000 description 1
- AWNAEZICPNGAJK-FXQIFTODSA-N Ala-Met-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(O)=O AWNAEZICPNGAJK-FXQIFTODSA-N 0.000 description 1
- DYXOFPBJBAHWFY-JBDRJPRFSA-N Ala-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N DYXOFPBJBAHWFY-JBDRJPRFSA-N 0.000 description 1
- GXXWTNKNFFKTJB-NAKRPEOUSA-N Arg-Ile-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O GXXWTNKNFFKTJB-NAKRPEOUSA-N 0.000 description 1
- ISJWBVIYRBAXEB-CIUDSAMLSA-N Arg-Ser-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O ISJWBVIYRBAXEB-CIUDSAMLSA-N 0.000 description 1
- FRBAHXABMQXSJQ-FXQIFTODSA-N Arg-Ser-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O FRBAHXABMQXSJQ-FXQIFTODSA-N 0.000 description 1
- SLKLLQWZQHXYSV-CIUDSAMLSA-N Asn-Ala-Lys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O SLKLLQWZQHXYSV-CIUDSAMLSA-N 0.000 description 1
- JBDLMLZNDRLDIX-HJGDQZAQSA-N Asn-Thr-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O JBDLMLZNDRLDIX-HJGDQZAQSA-N 0.000 description 1
- YQPSDMUGFKJZHR-QRTARXTBSA-N Asn-Trp-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC(=O)N)N YQPSDMUGFKJZHR-QRTARXTBSA-N 0.000 description 1
- JPPLRQVZMZFOSX-UWJYBYFXSA-N Asn-Tyr-Ala Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 JPPLRQVZMZFOSX-UWJYBYFXSA-N 0.000 description 1
- VAWNQIGQPUOPQW-ACZMJKKPSA-N Asp-Glu-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O VAWNQIGQPUOPQW-ACZMJKKPSA-N 0.000 description 1
- LIVXPXUVXFRWNY-CIUDSAMLSA-N Asp-Lys-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O LIVXPXUVXFRWNY-CIUDSAMLSA-N 0.000 description 1
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- NOCCABSVTRONIN-CIUDSAMLSA-N Cys-Ala-Leu Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CS)N NOCCABSVTRONIN-CIUDSAMLSA-N 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- JSYULGSPLTZDHM-NRPADANISA-N Gln-Ala-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O JSYULGSPLTZDHM-NRPADANISA-N 0.000 description 1
- MAGNEQBFSBREJL-DCAQKATOSA-N Gln-Glu-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)N MAGNEQBFSBREJL-DCAQKATOSA-N 0.000 description 1
- FQCILXROGNOZON-YUMQZZPRSA-N Gln-Pro-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O FQCILXROGNOZON-YUMQZZPRSA-N 0.000 description 1
- DUGYCMAIAKAQPB-GLLZPBPUSA-N Gln-Thr-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DUGYCMAIAKAQPB-GLLZPBPUSA-N 0.000 description 1
- VLOLPWWCNKWRNB-LOKLDPHHSA-N Gln-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O VLOLPWWCNKWRNB-LOKLDPHHSA-N 0.000 description 1
- SJJHXJDSNQJMMW-SRVKXCTJSA-N Glu-Lys-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O SJJHXJDSNQJMMW-SRVKXCTJSA-N 0.000 description 1
- BPLNJYHNAJVLRT-ACZMJKKPSA-N Glu-Ser-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O BPLNJYHNAJVLRT-ACZMJKKPSA-N 0.000 description 1
- YQPFCZVKMUVZIN-AUTRQRHGSA-N Glu-Val-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O YQPFCZVKMUVZIN-AUTRQRHGSA-N 0.000 description 1
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- MIIVFRCYJABHTQ-ONGXEEELSA-N Gly-Leu-Val Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O MIIVFRCYJABHTQ-ONGXEEELSA-N 0.000 description 1
- FKYQEVBRZSFAMJ-QWRGUYRKSA-N Gly-Ser-Tyr Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FKYQEVBRZSFAMJ-QWRGUYRKSA-N 0.000 description 1
- XHVONGZZVUUORG-WEDXCCLWSA-N Gly-Thr-Lys Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCCN XHVONGZZVUUORG-WEDXCCLWSA-N 0.000 description 1
- FOKISINOENBSDM-WLTAIBSBSA-N Gly-Thr-Tyr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O FOKISINOENBSDM-WLTAIBSBSA-N 0.000 description 1
- YGHSQRJSHKYUJY-SCZZXKLOSA-N Gly-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN YGHSQRJSHKYUJY-SCZZXKLOSA-N 0.000 description 1
- 101000619708 Homo sapiens Peroxiredoxin-6 Proteins 0.000 description 1
- ZLFNNVATRMCAKN-ZKWXMUAHSA-N Ile-Ser-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZLFNNVATRMCAKN-ZKWXMUAHSA-N 0.000 description 1
- YBKKLDBBPFIXBQ-MBLNEYKQSA-N Ile-Thr-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)O)N YBKKLDBBPFIXBQ-MBLNEYKQSA-N 0.000 description 1
- DZMWFIRHFFVBHS-ZEWNOJEFSA-N Ile-Tyr-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)O)N DZMWFIRHFFVBHS-ZEWNOJEFSA-N 0.000 description 1
- NXRNRBOKDBIVKQ-CXTHYWKRSA-N Ile-Tyr-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N NXRNRBOKDBIVKQ-CXTHYWKRSA-N 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- AXZGZMGRBDQTEY-SRVKXCTJSA-N Leu-Gln-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O AXZGZMGRBDQTEY-SRVKXCTJSA-N 0.000 description 1
- FEHQLKKBVJHSEC-SZMVWBNQSA-N Leu-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FEHQLKKBVJHSEC-SZMVWBNQSA-N 0.000 description 1
- YWKNKRAKOCLOLH-OEAJRASXSA-N Leu-Phe-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)O)C(O)=O)CC1=CC=CC=C1 YWKNKRAKOCLOLH-OEAJRASXSA-N 0.000 description 1
- KIZIOFNVSOSKJI-CIUDSAMLSA-N Leu-Ser-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N KIZIOFNVSOSKJI-CIUDSAMLSA-N 0.000 description 1
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 1
- AIQWYVFNBNNOLU-RHYQMDGZSA-N Leu-Thr-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O AIQWYVFNBNNOLU-RHYQMDGZSA-N 0.000 description 1
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 1
- GQZMPWBZQALKJO-UWVGGRQHSA-N Lys-Gly-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O GQZMPWBZQALKJO-UWVGGRQHSA-N 0.000 description 1
- 229930126263 Maytansine Natural products 0.000 description 1
- GHQFLTYXGUETFD-UFYCRDLUSA-N Met-Tyr-Tyr Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N GHQFLTYXGUETFD-UFYCRDLUSA-N 0.000 description 1
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 102100022239 Peroxiredoxin-6 Human genes 0.000 description 1
- NAXPHWZXEXNDIW-JTQLQIEISA-N Phe-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 NAXPHWZXEXNDIW-JTQLQIEISA-N 0.000 description 1
- YDUGVDGFKNXFPL-IXOXFDKPSA-N Phe-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N)O YDUGVDGFKNXFPL-IXOXFDKPSA-N 0.000 description 1
- FGWUALWGCZJQDJ-URLPEUOOSA-N Phe-Thr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGWUALWGCZJQDJ-URLPEUOOSA-N 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 229920002535 Polyethylene Glycol 1500 Polymers 0.000 description 1
- KIGGUSRFHJCIEJ-DCAQKATOSA-N Pro-Asp-His Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O KIGGUSRFHJCIEJ-DCAQKATOSA-N 0.000 description 1
- ZCXQTRXYZOSGJR-FXQIFTODSA-N Pro-Asp-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O ZCXQTRXYZOSGJR-FXQIFTODSA-N 0.000 description 1
- QEDMOZUJTGEIBF-FXQIFTODSA-N Ser-Arg-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O QEDMOZUJTGEIBF-FXQIFTODSA-N 0.000 description 1
- ICHZYBVODUVUKN-SRVKXCTJSA-N Ser-Asn-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ICHZYBVODUVUKN-SRVKXCTJSA-N 0.000 description 1
- UIGMAMGZOJVTDN-WHFBIAKZSA-N Ser-Gly-Ser Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O UIGMAMGZOJVTDN-WHFBIAKZSA-N 0.000 description 1
- UBRMZSHOOIVJPW-SRVKXCTJSA-N Ser-Leu-Lys Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O UBRMZSHOOIVJPW-SRVKXCTJSA-N 0.000 description 1
- RHAPJNVNWDBFQI-BQBZGAKWSA-N Ser-Pro-Gly Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O RHAPJNVNWDBFQI-BQBZGAKWSA-N 0.000 description 1
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 1
- CJEHCEOXPLASCK-MEYUZBJRSA-N Thr-Tyr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@H](O)C)CC1=CC=C(O)C=C1 CJEHCEOXPLASCK-MEYUZBJRSA-N 0.000 description 1
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 1
- KZTLZZQTJMCGIP-ZJDVBMNYSA-N Thr-Val-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KZTLZZQTJMCGIP-ZJDVBMNYSA-N 0.000 description 1
- 101710120037 Toxin CcdB Proteins 0.000 description 1
- ZJPSMXCFEKMZFE-IHPCNDPISA-N Trp-Tyr-Ser Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O ZJPSMXCFEKMZFE-IHPCNDPISA-N 0.000 description 1
- MBLJBGZWLHTJBH-SZMVWBNQSA-N Trp-Val-Arg Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)=CNC2=C1 MBLJBGZWLHTJBH-SZMVWBNQSA-N 0.000 description 1
- 229940122429 Tubulin inhibitor Drugs 0.000 description 1
- SOAUMCDLIUGXJJ-SRVKXCTJSA-N Tyr-Ser-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O SOAUMCDLIUGXJJ-SRVKXCTJSA-N 0.000 description 1
- UQMPYVLTQCGRSK-IFFSRLJSSA-N Val-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N)O UQMPYVLTQCGRSK-IFFSRLJSSA-N 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 238000011091 antibody purification Methods 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- WKPWGQKGSOKKOO-RSFHAFMBSA-N maytansine Chemical compound CO[C@@H]([C@@]1(O)C[C@](OC(=O)N1)([C@H]([C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(C)=O)CC(=O)N1C)C)[H])\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 WKPWGQKGSOKKOO-RSFHAFMBSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 1
- 108010070643 prolylglutamic acid Proteins 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 231100000057 systemic toxicity Toxicity 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 108010044292 tryptophyltyrosine Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Landscapes
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明提供了一种抗DM1单克隆抗体及其制备方法与应用，所述抗体包含有重链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，轻链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3。本发明的所述抗DM1单克隆抗体可以应用于相关抗肿瘤配体偶联药物中的研究。
Description
技术领域
本发明属于生物医药技术领域，具体涉及一种抗DM1单克隆抗体及其制备方法与应用。
背景技术
Mertansine (Synonyms: DM1; Maytansinoid DM1)是一种微管蛋白抑制剂，也是一种抗体可缀合的美登木素生物碱，用于克服与美登素相关的全身毒性并增强肿瘤特异性递送。Mertansine可以通过连接体连接到单克隆抗体上，形成抗体偶联药物(ADC)。在肿瘤的治疗中，具有抗癌活性的小分子化合物，由于其对正常组织的毒性而限制了其使用，而将其与靶向配体（单克隆抗体、多肽、融合蛋白等）结合，将活性分子带到靶区域，可以有效地控制小分子化合物对于正常细胞产生毒性的问题。因此，配体药物偶联物，如ADC(AntibodyDrug Conjugate，ADC)，PDC(peptide drug conjugate)等XDC应运而生。如何合成一种抗DM1单克隆抗体，以更好的针对相关抗体偶联药物，成为现在需要解决的问题。
发明内容
为了解决现有技术的不足，本发明提供了一种抗DM1单克隆抗体及其制备与应用。
本发明的目的通过以下技术方案来实现：
抗DM1单克隆抗体，包含有重链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，其氨基酸序列分别为SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3；
轻链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，其氨基酸序列分别为SEQID NO:4、SEQ ID NO:5、SEQ ID NO:6。
优选地，所述抗体具有SEQ ID NO：7所示的重链区可变区氨基酸序列和SEQ IDNO:8所示的轻链区可变区氨基酸序列。
一种核酸分子，所述核酸分子包括编码如权利要求1-2任一所述的抗DM1单克隆抗体。
一种细胞系，能够产生以上任一所述的抗DM1单克隆抗体。
抗DM1单克隆抗体的制备方法，所述方法包括，培养以上所述的细胞系，并产生所述的抗DM1单克隆抗体。
抗DM1单克隆抗体的应用，所述抗DM1单克隆抗体在制备治疗和预防相关抗肿瘤配体偶联药物中的应用，所述配体可以是抗体，多肽，融合蛋白等。
本发明的有益效果体现在：提供了一种抗DM1单克隆抗体，并提供了相应的制备方法，本发明的所述抗DM1单克隆抗体可以应用于在相关抗肿瘤配体偶联药物中的研究。
附图说明
图1：本发明实施例5中抗体与抗原结合在450 nm波长下的OD值。
具体实施方式
本发明揭示了一种抗DM1单克隆抗体，包含有重链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，其氨基酸序列分别为SEQ ID NO:1 GFTFSSYA；SEQ ID NO:2ISGGGSYI；SEQ ID NO:3 ARQGYGYDGSFGFDF。轻链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，其氨基酸序列分别为SEQ ID NO:4 IGAVTTSNY；SEQ ID NO:5 GTY；SEQ ID NO:6ALWYSNHWV。
所述抗体具有SEQ ID NO:7所示的重链区可变区氨基酸序列和SEQ ID NO:8所示的轻链区可变区氨基酸序列。
其中，重链可变区,SEQ ID NO:7
EVQLVESGGGLVQPGGSLKLSCAASGFTFSSYAMSWVRQTPEKRLEWVASISGGGSYIYYPDSVKGRFTISRDNAKNTLYLQMSSLRSEDTAMYYCARQGYGYDGSFGFDFWGQGTLVTVSA轻链可变区，SEQ ID NO:8QAVVTQESALTTSPGETVTFTCRSSIGAVTTSNYANWVQEKPDHLFTGLIGGTYKRISGVPARFSGSLIGDKAALTITGAQTEDEAIYFCALWYSNHWVFGGGTKLTVL
本发明还揭示了一种核酸分子及细胞系，以及以上所述抗DM1单克隆抗体的制备方法。
所述核酸分子由编码以上任一所述的抗DM1单克隆抗体获得。所述细胞系能够产生如以上任一所述的抗DM1单克隆抗体。所述方法包括培养上述的细胞系，并产生所述的抗DM1单克隆抗体。本发明的所述抗DM1单克隆抗体可以应用于相关抗肿瘤配体偶联药物中的研究。
为更好的理解本发明，以下进行具体的实施举例说明。
实施例1：抗原制备
公司内部合成DM1小分子，并标记一个半胱氨酸（DM1-CYS）以便于偶联载体蛋白，进行免疫或检测用。载体蛋白偶联，免疫抗原制备，由于DM1分子比较小，毒性大，不适合直接免疫，将DM1-CYS与载体蛋白进行偶联，选择Thermo Scientific的马来酰胺活化的钥孔血蓝蛋白（KLH）试剂盒，按照试剂盒提供的流程操作，进行与载体蛋白偶联。
实施例2：动物免疫
将实施例1中制备的免疫用抗原（DM1-CYS-KLH）用弗氏完全佐剂乳化，免疫4~6周龄雌性Balb/c小鼠（购自北京维通利华实验动物技术有限公司），皮下注射，每只小鼠5~6点，剂量为100µg/只。每14天加强免疫一次，抗原使用弗氏非完全佐剂乳化，剂量为50µg/只。第3次加强免疫后7天，以间接ELISA（波长450nm）检测小鼠血清中抗免疫原的多抗效价，效价最高的小鼠以尾静脉注射冲击免疫，抗原用生理盐水混匀，剂量为50µg/只。
实施例3：杂交瘤细胞制备
1、融合：将实施例2中免疫达标的小鼠采血，取脾脏制备细胞悬液，与小鼠骨髓瘤细胞F0以10：1比例混合，离心1500rpm，5分钟。弃上清后，离心管放入37℃水浴中，在1分钟内缓慢加入1 mL的PEG1500（Roche公司），并搅动细胞。在温水中静置l分钟后，加入10 mL无血清的IMDM（Hyclone公司），混匀，离心1000rpm，5分钟。弃上清后，加入含有10 %胎牛血清（Excell公司）的培养基，小心的将细胞吹打起来，并加入配好的HAT完全培养基（Sigma公司）混匀，铺96孔板。将铺好细胞的培养板，放入37℃、5% CO2培养箱中培养。
2、筛选及亚克隆：利用ELISA的方法进行初步的筛选，将筛选得到的阳性抗体进行稀释直至得到单个亚克隆细胞。通过ELISA筛选，得到多个阳性克隆，挑选其中一个阳性克隆进行扩大培养，检测定株。以下以编号06CT8.1.1.1为挑选的个体举例说明。
实施例4：腹水诱生法制备单克隆抗体
1、腹水制备
对数生长期细胞用无血清培养基洗涤并悬起，计数3-7x105/mL，1mL悬浮的细胞腹腔注射事先用石蜡油致敏的小鼠。小鼠腹部鼓胀后开始收集腹水，可多次收取。取出的腹水于4℃下离心4000rpm，10分钟。小心吸出中间的腹水收集于离心管中，4℃或-20℃保存。
2、单克隆抗体纯化
用Protein G（GE公司）亲和层析法按说明书从腹水中纯化抗体。SDS-PAGE胶鉴定纯度，Bradford法测定浓度。纯化的抗体保存于-20℃。
实施例5：单克隆抗体特性鉴定
1、亚型检测：选用Mouse Immunoglobulin Isotyping ELISA试剂盒（BD），按操作说明，羊抗鼠IgG板子包被、封闭。取杂交瘤上清，进行检测。结果显示，克隆06CT8.1.1.1为IgG1 λ型鼠源单克隆抗体。
2.抗体效价检测
用PBS（pH 7.2）稀释包被BSA- DM1，1µg/mL，每孔加100 µL，4℃，过夜。倾空液体，用含0.05% Tween的PBS洗3次，每孔加入200 µL封闭液（含3% BSA的PBS），37℃下孵育1小时，倾空液体，备用。
每孔加入稀释好的抗体溶液，1:3做梯度稀释，每孔100 µL反应液，37℃下孵育1小时，倾空液体，用PBST清洗3次。用封闭液1:10000稀释HRP标记的羊抗鼠抗体Anti-MouseIgG(Fc specific)（sigma公司），每孔分别加入100 µL，37℃下孵育1小时，倾空液体，用PBST清洗4次。每孔加100µLTMB显色液进行显色反应，10-20分钟内测定450 nm波长下的OD值。结合图1所示，显示挑选的单抗具有与DM1较好的结合活性。
实施例6：抗体可变区序列测定
取培养的杂交瘤细胞系1x106，离心取上清，将细胞沉淀委托苏州金唯智生物科技有限公司进行抗体重链和轻链可变区序列测定，如下示出示例性的抗体重链可变区序列和抗体轻链可变区序列。
抗体CDR氨基酸序列
| 样品编号 | 链型 | aaCDR1 | aaCDR2 | aaCDR3 |
| 06CT8.1.1.1-VH | IGH | GFTFSSYA | ISGGGSYI | ARQGYGYDGSFGFDF |
| 06CT8.1.1.1-VL | IGK | IGAVTTSNY | GTY | ALWYSNHWV |
抗体可变区氨基酸序列
重链可变区，SEQ ID NO:7
EVQLVESGGGLVQPGGSLKLSCAASGFTFSSYAMSWVRQTPEKRLEWVASISGGGSYIYYPDSVKGRFTISRDNAKNTLYLQMSSLRSEDTAMYYCARQGYGYDGSFGFDFWGQGTLVTVSA
轻链可变区，SEQ ID NO:8
QAVVTQESALTTSPGETVTFTCRSSIGAVTTSNYANWVQEKPDHLFTGLIGGTYKRISGVPARFSGSLIGDKAALTITGAQTEDEAIYFCALWYSNHWVFGGGTKLTVL
编码抗体可变区的核酸序列
重链可变区，SEQ ID NO:9
GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGCAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAGAAGAGGCTGGAGTGGGTCGCATCCATTAGTGGCGGTGGTAGTTACATTTACTATCCTGACAGTGTGAAGGGTCGATTCACCATCTCCAGAGACAATGCCAAGAATACCCTGTACCTACAAATGAGCAGTCTGAGGTCTGAGGACACGGCCATGTATTACTGTGCAAGACAAGGCTATGGCTATGATGGTTCCTTCGGGTTTGATTTCTGGGGCCAGGGGACTCTGGTTACTGTCTCTGCA
轻链可变区，SEQ ID NO:10
CAGGCTGTTGTGACTCAGGAATCTGCACTCACCACATCACCTGGTGAAACAGTCACATTCACTTGTCGCTCAAGTATTGGGGCTGTTACAACTAGTAACTATGCCAACTGGGTCCAAGAAAAACCAGATCATTTATTCACTGGTCTAATAGGTGGTACCTACAAACGAATTTCAGGTGTTCCTGCCAGATTCTCAGGCTCCCTGATTGGAGACAAGGCTGCCCTCACCATCACAGGGGCACAGACTGAGGATGAGGCAATATATTTCTGTGCTCTATGGTACAGCAACCATTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA。
且以上实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的精神和范围。
序列表
<110> 苏州宜百奥生物科技有限公司
<120> 抗DM1单克隆抗体及其制备方法与应用
<160> 8
<170> SIPOSequenceListing 1.0
<210> 1
<211> 8
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 1
Gly Phe Thr Phe Ser Ser Tyr Ala
1 5
<210> 2
<211> 8
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 2
Ile Ser Gly Gly Gly Ser Tyr Ile
1 5
<210> 3
<211> 15
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 3
Ala Arg Gln Gly Tyr Gly Tyr Asp Gly Ser Phe Gly Phe Asp Phe
1 5 10 15
<210> 4
<211> 9
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 4
Ile Gly Ala Val Thr Thr Ser Asn Tyr
1 5
<210> 5
<211> 3
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 5
Gly Thr Tyr
1
<210> 6
<211> 9
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 6
Ala Leu Trp Tyr Ser Asn His Trp Val
1 5
<210> 7
<211> 122
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 7
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val
35 40 45
Ala Ser Ile Ser Gly Gly Gly Ser Tyr Ile Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Gln Gly Tyr Gly Tyr Asp Gly Ser Phe Gly Phe Asp Phe Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ala
115 120
<210> 8
<211> 109
<212> PRT
<213> 抗体可变区序列(鼠源抗体)
<400> 8
Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu
1 5 10 15
Thr Val Thr Phe Thr Cys Arg Ser Ser Ile Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly
35 40 45
Leu Ile Gly Gly Thr Tyr Lys Arg Ile Ser Gly Val Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala
65 70 75 80
Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
Claims (6)
1.抗DM1单克隆抗体，其特征在于：包含有重链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，其氨基酸序列分别为SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3；
轻链可变区的抗原互补决定区aaCDR1、aaCDR2和aaCDR3，其氨基酸序列分别为SEQ IDNO:4、SEQ ID NO:5、SEQ ID NO:6。
2.如权利要求1所述的抗DM1单克隆抗体，其特征在于：所述抗体具有SEQ ID NO：7所示的重链区可变区氨基酸序列和SEQ ID NO:8所示的轻链区可变区氨基酸序列。
3.一种核酸分子，其特征在于：所述核酸分子包括编码如权利要求1-2任一所述的抗DM1单克隆抗体。
4.一种细胞系，其特征在于：能够产生如权利要求1-2任一所述的抗DM1单克隆抗体。
5.抗DM1单克隆抗体的制备方法，其特征在于：所述方法包括，培养如权利要求4所述的细胞系，并产生所述的抗DM1单克隆抗体。
6.抗DM1单克隆抗体的应用，其特征在于：所述抗DM1单克隆抗体在相关抗肿瘤配体偶联药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111577267.3A CN114276448A (zh) | 2021-12-22 | 2021-12-22 | 抗dm1单克隆抗体及其制备方法与应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111577267.3A CN114276448A (zh) | 2021-12-22 | 2021-12-22 | 抗dm1单克隆抗体及其制备方法与应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114276448A true CN114276448A (zh) | 2022-04-05 |
Family
ID=80873894
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111577267.3A Withdrawn CN114276448A (zh) | 2021-12-22 | 2021-12-22 | 抗dm1单克隆抗体及其制备方法与应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114276448A (zh) |
-
2021
- 2021-12-22 CN CN202111577267.3A patent/CN114276448A/zh not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109206514B (zh) | Tslp单克隆抗体及其制备方法和应用 | |
| JP7209464B2 (ja) | ヒトインターロイキン－２に対する免疫刺激性モノクローナル抗体 | |
| CN109734805B (zh) | 抗ck20蛋白单克隆抗体、细胞系及其制备方法和应用 | |
| CN110218251B (zh) | 抗msh2蛋白单克隆抗体、细胞系及其制备方法和应用 | |
| CN110563843A (zh) | 一种靶向人vista蛋白的单克隆抗体 | |
| CN116529265A (zh) | Cd73的抗原结合蛋白及其应用 | |
| CN111363037B (zh) | 一种包含特异性结合afp蛋白的抗体的疾病检测试剂盒 | |
| CN110642950B (zh) | 一种人源化t细胞活化的v域免疫抑制因子抗原结合片段 | |
| CN114195897B (zh) | Pd-l1单克隆抗体、重链、轻链可变区、单克隆细胞株及运用和试剂盒 | |
| CN116396387A (zh) | Pd-l1单克隆抗体、重链、轻链可变区、单克隆细胞株及运用和试剂盒 | |
| CN114276448A (zh) | 抗dm1单克隆抗体及其制备方法与应用 | |
| CN111558038B (zh) | 一种用于癌症治疗的免疫检查点抑制剂 | |
| CN115505043A (zh) | 特异性结合糖基化ceacam5的抗体 | |
| CN110713538A (zh) | 一种抗人cd28单克隆抗体及其用途 | |
| CN110579610A (zh) | 用于检测t细胞活化的v域免疫抑制因子的试剂盒 | |
| CN114181307A (zh) | 抗mmae单克隆抗体及其制备方法与应用 | |
| CN114276455A (zh) | 抗HIgG-Fc单克隆抗体及其制备方法与应用 | |
| CN114163528B (zh) | Cd47结合分子及其应用 | |
| CN113321730B (zh) | Cldn18.2抗体及其应用 | |
| CN112724253B (zh) | 抗人穹窿体蛋白的抗体及其应用 | |
| WO2023134716A1 (zh) | 一种结合b7h3和nkp30的双特异性抗体及其应用 | |
| CN117384290B (zh) | 人源robo1结合分子及其应用 | |
| CN111875697B (zh) | 一种包含肽标签的融合蛋白 | |
| CN113929783B (zh) | 抗cd99蛋白单克隆抗体、细胞系及其制备方法和应用 | |
| CN112626009B (zh) | 一种调整干细胞成软骨分化的方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220405 |