CN114478779A - 人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用 - Google Patents
人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用 Download PDFInfo
- Publication number
- CN114478779A CN114478779A CN202011158008.2A CN202011158008A CN114478779A CN 114478779 A CN114478779 A CN 114478779A CN 202011158008 A CN202011158008 A CN 202011158008A CN 114478779 A CN114478779 A CN 114478779A
- Authority
- CN
- China
- Prior art keywords
- seq
- amino acid
- antibody
- gly
- ser
- 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.)
- Granted
Links
- 239000003814 drug Substances 0.000 claims abstract description 12
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 58
- 125000000539 amino acid group Chemical group 0.000 claims description 44
- 108090000623 proteins and genes Proteins 0.000 claims description 22
- 102000004169 proteins and genes Human genes 0.000 claims description 14
- 102000010956 Glypican Human genes 0.000 claims description 12
- 108050001154 Glypican Proteins 0.000 claims description 12
- 108050007237 Glypican-3 Proteins 0.000 claims description 12
- 206010028980 Neoplasm Diseases 0.000 claims description 11
- 239000013598 vector Substances 0.000 claims description 11
- 229940127121 immunoconjugate Drugs 0.000 claims description 10
- 239000002157 polynucleotide Substances 0.000 claims description 10
- 102000040430 polynucleotide Human genes 0.000 claims description 10
- 108091033319 polynucleotide Proteins 0.000 claims description 10
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims description 9
- 239000003053 toxin Substances 0.000 claims description 9
- 231100000765 toxin Toxicity 0.000 claims description 9
- 108700012359 toxins Proteins 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 8
- 101000762949 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) Exotoxin A Proteins 0.000 claims description 7
- 201000011510 cancer Diseases 0.000 claims description 7
- 231100000433 cytotoxic Toxicity 0.000 claims description 6
- 230000001472 cytotoxic effect Effects 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 5
- 230000000259 anti-tumor effect Effects 0.000 claims description 4
- 239000012636 effector Substances 0.000 claims description 4
- MFRNYXJJRJQHNW-DEMKXPNLSA-N (2s)-2-[[(2r,3r)-3-methoxy-3-[(2s)-1-[(3r,4s,5s)-3-methoxy-5-methyl-4-[methyl-[(2s)-3-methyl-2-[[(2s)-3-methyl-2-(methylamino)butanoyl]amino]butanoyl]amino]heptanoyl]pyrrolidin-2-yl]-2-methylpropanoyl]amino]-3-phenylpropanoic acid Chemical compound CN[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MFRNYXJJRJQHNW-DEMKXPNLSA-N 0.000 claims description 3
- 208000036142 Viral infection Diseases 0.000 claims description 3
- IEDXPSOJFSVCKU-HOKPPMCLSA-N [4-[[(2S)-5-(carbamoylamino)-2-[[(2S)-2-[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-3-[[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate Chemical compound CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)c1ccccc1)OC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCc1ccc(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN2C(=O)CCC2=O)C(C)C)cc1)C(C)C IEDXPSOJFSVCKU-HOKPPMCLSA-N 0.000 claims description 3
- 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 claims description 3
- 108010093470 monomethyl auristatin E Proteins 0.000 claims description 3
- 108010059074 monomethylauristatin F Proteins 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 230000009385 viral infection Effects 0.000 claims description 3
- 108010066676 Abrin Proteins 0.000 claims description 2
- 101100450694 Arabidopsis thaliana HFR1 gene Proteins 0.000 claims description 2
- 208000023275 Autoimmune disease Diseases 0.000 claims description 2
- 108030001720 Bontoxilysin Proteins 0.000 claims description 2
- 102000016607 Diphtheria Toxin Human genes 0.000 claims description 2
- 108010053187 Diphtheria Toxin Proteins 0.000 claims description 2
- 108010039491 Ricin Proteins 0.000 claims description 2
- 229940053031 botulinum toxin Drugs 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 229940126585 therapeutic drug Drugs 0.000 claims description 2
- 239000000032 diagnostic agent Substances 0.000 claims 1
- 229940039227 diagnostic agent Drugs 0.000 claims 1
- 125000002576 diazepinyl group Chemical class N1N=C(C=CC=C1)* 0.000 claims 1
- 239000003550 marker Substances 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 102100032530 Glypican-3 Human genes 0.000 abstract description 32
- 101001014668 Homo sapiens Glypican-3 Proteins 0.000 abstract description 32
- 230000005847 immunogenicity Effects 0.000 abstract description 8
- 230000000694 effects Effects 0.000 abstract description 5
- 230000002637 immunotoxin Effects 0.000 abstract description 3
- 229940051026 immunotoxin Drugs 0.000 abstract description 3
- 239000002596 immunotoxin Substances 0.000 abstract description 3
- 231100000608 immunotoxin Toxicity 0.000 abstract description 3
- 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 32
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 31
- 210000004027 cell Anatomy 0.000 description 30
- 241000287828 Gallus gallus Species 0.000 description 24
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 22
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 21
- 230000027455 binding Effects 0.000 description 13
- VKKYFICVTYKFIO-CIUDSAMLSA-N Arg-Ala-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N VKKYFICVTYKFIO-CIUDSAMLSA-N 0.000 description 12
- 235000018102 proteins Nutrition 0.000 description 12
- NINQYGGNRIBFSC-CIUDSAMLSA-N Ala-Lys-Ser Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CO)C(O)=O NINQYGGNRIBFSC-CIUDSAMLSA-N 0.000 description 11
- YHBDGLZYNIARKJ-GUBZILKMSA-N Ala-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N YHBDGLZYNIARKJ-GUBZILKMSA-N 0.000 description 11
- PGNNQOJOEGFAOR-KWQFWETISA-N Ala-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=C(O)C=C1 PGNNQOJOEGFAOR-KWQFWETISA-N 0.000 description 11
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 description 11
- CUQDCPXNZPDYFQ-ZLUOBGJFSA-N Asp-Ser-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O CUQDCPXNZPDYFQ-ZLUOBGJFSA-N 0.000 description 11
- 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 11
- XTHUKRLJRUVVBF-WHFBIAKZSA-N Cys-Gly-Ser Chemical compound SC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O XTHUKRLJRUVVBF-WHFBIAKZSA-N 0.000 description 11
- OYTPNWYZORARHL-XHNCKOQMSA-N Gln-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N OYTPNWYZORARHL-XHNCKOQMSA-N 0.000 description 11
- ALOBJFDJTMQQPW-ONGXEEELSA-N Gly-His-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)CN ALOBJFDJTMQQPW-ONGXEEELSA-N 0.000 description 11
- PDUHNKAFQXQNLH-ZETCQYMHSA-N Gly-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)NCC(O)=O PDUHNKAFQXQNLH-ZETCQYMHSA-N 0.000 description 11
- 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 11
- YOBGUCWZPXJHTN-BQBZGAKWSA-N Gly-Ser-Arg Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YOBGUCWZPXJHTN-BQBZGAKWSA-N 0.000 description 11
- 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 11
- RZEDHGORCKRINR-STQMWFEESA-N Gly-Trp-Cys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN RZEDHGORCKRINR-STQMWFEESA-N 0.000 description 11
- RPZFUIQVAPZLRH-GHCJXIJMSA-N Ile-Asp-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C)C(=O)O)N RPZFUIQVAPZLRH-GHCJXIJMSA-N 0.000 description 11
- UAELWXJFLZBKQS-WHOFXGATSA-N Ile-Phe-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)NCC(O)=O UAELWXJFLZBKQS-WHOFXGATSA-N 0.000 description 11
- RKQAYOWLSFLJEE-SVSWQMSJSA-N Ile-Thr-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)O)N RKQAYOWLSFLJEE-SVSWQMSJSA-N 0.000 description 11
- 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 11
- 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 11
- 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 11
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 11
- HXSUFWQYLPKEHF-IHRRRGAJSA-N Phe-Asn-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N HXSUFWQYLPKEHF-IHRRRGAJSA-N 0.000 description 11
- BPCLGWHVPVTTFM-QWRGUYRKSA-N Phe-Ser-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)NCC(O)=O BPCLGWHVPVTTFM-QWRGUYRKSA-N 0.000 description 11
- 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 11
- SQBLRDDJTUJDMV-ACZMJKKPSA-N Ser-Glu-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQBLRDDJTUJDMV-ACZMJKKPSA-N 0.000 description 11
- HAYADTTXNZFUDM-IHRRRGAJSA-N Ser-Tyr-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O HAYADTTXNZFUDM-IHRRRGAJSA-N 0.000 description 11
- DCLBXIWHLVEPMQ-JRQIVUDYSA-N Thr-Asp-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 DCLBXIWHLVEPMQ-JRQIVUDYSA-N 0.000 description 11
- 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 11
- DANHCMVVXDXOHN-SRVKXCTJSA-N Tyr-Asp-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 DANHCMVVXDXOHN-SRVKXCTJSA-N 0.000 description 11
- ZLFHAAGHGQBQQN-AEJSXWLSSA-N Val-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N ZLFHAAGHGQBQQN-AEJSXWLSSA-N 0.000 description 11
- ZLFHAAGHGQBQQN-GUBZILKMSA-N Val-Ala-Pro Natural products CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O ZLFHAAGHGQBQQN-GUBZILKMSA-N 0.000 description 11
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(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 OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 11
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 11
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 11
- 108010008355 arginyl-glutamine Proteins 0.000 description 11
- 108010016616 cysteinylglycine Proteins 0.000 description 11
- 108010068404 exorphin B4 Proteins 0.000 description 11
- 108010089804 glycyl-threonine Proteins 0.000 description 11
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 11
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 11
- 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 10
- LMFXXZPPZDCPTA-ZKWXMUAHSA-N Ala-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N LMFXXZPPZDCPTA-ZKWXMUAHSA-N 0.000 description 10
- ALMIMUZAWTUNIO-BZSNNMDCSA-N Asp-Tyr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ALMIMUZAWTUNIO-BZSNNMDCSA-N 0.000 description 10
- ZEEPYMXTJWIMSN-GUBZILKMSA-N Gln-Lys-Ser Chemical compound NCCCC[C@@H](C(=O)N[C@@H](CO)C(O)=O)NC(=O)[C@@H](N)CCC(N)=O ZEEPYMXTJWIMSN-GUBZILKMSA-N 0.000 description 10
- 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 10
- XXCDTYBVGMPIOA-FXQIFTODSA-N Glu-Asp-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O XXCDTYBVGMPIOA-FXQIFTODSA-N 0.000 description 10
- 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 10
- PYTZFYUXZZHOAD-WHFBIAKZSA-N Gly-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)CN PYTZFYUXZZHOAD-WHFBIAKZSA-N 0.000 description 10
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 10
- 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 10
- ZZWUYQXMIFTIIY-WEDXCCLWSA-N Gly-Thr-Leu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O ZZWUYQXMIFTIIY-WEDXCCLWSA-N 0.000 description 10
- 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 10
- 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 10
- 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 10
- LCNASHSOFMRYFO-WDCWCFNPSA-N Leu-Thr-Gln Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O LCNASHSOFMRYFO-WDCWCFNPSA-N 0.000 description 10
- FUKDBQGFSJUXGX-RWMBFGLXSA-N Lys-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)N)C(=O)O FUKDBQGFSJUXGX-RWMBFGLXSA-N 0.000 description 10
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 10
- JMVQDLDPDBXAAX-YUMQZZPRSA-N Pro-Gly-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 JMVQDLDPDBXAAX-YUMQZZPRSA-N 0.000 description 10
- QYSFWUIXDFJUDW-DCAQKATOSA-N Ser-Leu-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYSFWUIXDFJUDW-DCAQKATOSA-N 0.000 description 10
- ILUOMMDDGREELW-OSUNSFLBSA-N Thr-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O ILUOMMDDGREELW-OSUNSFLBSA-N 0.000 description 10
- SVGAWGVHFIYAEE-JSGCOSHPSA-N Trp-Gly-Gln Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O)=CNC2=C1 SVGAWGVHFIYAEE-JSGCOSHPSA-N 0.000 description 10
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-GUBZILKMSA-N 0.000 description 10
- HTONZBWRYUKUKC-RCWTZXSCSA-N Val-Thr-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O HTONZBWRYUKUKC-RCWTZXSCSA-N 0.000 description 10
- 239000012634 fragment Substances 0.000 description 10
- 210000004602 germ cell Anatomy 0.000 description 10
- 108010078144 glutaminyl-glycine Proteins 0.000 description 10
- 108010049041 glutamylalanine Proteins 0.000 description 10
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 10
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 10
- 108010026333 seryl-proline Proteins 0.000 description 10
- LGFCAXJBAZESCF-ACZMJKKPSA-N Ala-Gln-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O LGFCAXJBAZESCF-ACZMJKKPSA-N 0.000 description 9
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 9
- VNFSAYFQLXPHPY-CIQUZCHMSA-N Ala-Thr-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNFSAYFQLXPHPY-CIQUZCHMSA-N 0.000 description 9
- 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 9
- KPSHWSWFPUDEGF-FXQIFTODSA-N Asp-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(O)=O KPSHWSWFPUDEGF-FXQIFTODSA-N 0.000 description 9
- KVGPYKUIHZJWGA-BQBZGAKWSA-N Cys-Met-Gly Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O KVGPYKUIHZJWGA-BQBZGAKWSA-N 0.000 description 9
- UMRIXLHPZZIOML-OALUTQOASA-N Gly-Trp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)CN UMRIXLHPZZIOML-OALUTQOASA-N 0.000 description 9
- 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 9
- OIQSIMFSVLLWBX-VOAKCMCISA-N Lys-Leu-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OIQSIMFSVLLWBX-VOAKCMCISA-N 0.000 description 9
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 9
- GOMUXSCOIWIJFP-GUBZILKMSA-N Pro-Ser-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GOMUXSCOIWIJFP-GUBZILKMSA-N 0.000 description 9
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 9
- MUJQWSAWLLRJCE-KATARQTJSA-N Ser-Leu-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MUJQWSAWLLRJCE-KATARQTJSA-N 0.000 description 9
- GYDFRTRSSXOZCR-ACZMJKKPSA-N Ser-Ser-Glu Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O GYDFRTRSSXOZCR-ACZMJKKPSA-N 0.000 description 9
- XGFYGMKZKFRGAI-RCWTZXSCSA-N Thr-Val-Arg Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N XGFYGMKZKFRGAI-RCWTZXSCSA-N 0.000 description 9
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 9
- NCXTYSVDWLAQGZ-ZKWXMUAHSA-N Asn-Ser-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O NCXTYSVDWLAQGZ-ZKWXMUAHSA-N 0.000 description 8
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 8
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 239000000427 antigen Substances 0.000 description 7
- 108091007433 antigens Proteins 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 5
- 229940024606 amino acid Drugs 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 238000006467 substitution reaction Methods 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 4
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 4
- 210000004899 c-terminal region Anatomy 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000000539 dimer Substances 0.000 description 4
- 201000007270 liver cancer Diseases 0.000 description 4
- 208000014018 liver neoplasm Diseases 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 229920002971 Heparan sulfate Polymers 0.000 description 3
- VRUFCJZQDACGLH-UVOCVTCTSA-N Thr-Leu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VRUFCJZQDACGLH-UVOCVTCTSA-N 0.000 description 3
- 102000005936 beta-Galactosidase Human genes 0.000 description 3
- 108010005774 beta-Galactosidase Proteins 0.000 description 3
- 239000012472 biological sample Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000003114 enzyme-linked immunosorbent spot assay Methods 0.000 description 3
- -1 etc.) Substances 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 3
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 210000004962 mammalian cell Anatomy 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000002823 phage display Methods 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 238000003118 sandwich ELISA Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 2
- HJGUQJJJXQGXGJ-FXQIFTODSA-N Cys-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N HJGUQJJJXQGXGJ-FXQIFTODSA-N 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- 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 2
- 108010004729 Phycoerythrin Proteins 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 2
- IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 description 2
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010494 dissociation reaction Methods 0.000 description 2
- 230000005593 dissociations Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 108091006047 fluorescent proteins Proteins 0.000 description 2
- 102000034287 fluorescent proteins Human genes 0.000 description 2
- 229930004094 glycosylphosphatidylinositol Natural products 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 230000002163 immunogen Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 238000004393 prognosis Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000002864 sequence alignment Methods 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 108091005957 yellow fluorescent proteins Proteins 0.000 description 2
- LRANPJDWHYRCER-UHFFFAOYSA-N 1,2-diazepine Chemical compound N1C=CC=CC=N1 LRANPJDWHYRCER-UHFFFAOYSA-N 0.000 description 1
- NVKAWKQGWWIWPM-ABEVXSGRSA-N 17-β-hydroxy-5-α-Androstan-3-one Chemical compound C1C(=O)CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 NVKAWKQGWWIWPM-ABEVXSGRSA-N 0.000 description 1
- CYDQOEWLBCCFJZ-UHFFFAOYSA-N 4-(4-fluorophenyl)oxane-4-carboxylic acid Chemical compound C=1C=C(F)C=CC=1C1(C(=O)O)CCOCC1 CYDQOEWLBCCFJZ-UHFFFAOYSA-N 0.000 description 1
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 1
- 101710191936 70 kDa protein Proteins 0.000 description 1
- OBVSBEYOMDWLRJ-BFHQHQDPSA-N Ala-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N OBVSBEYOMDWLRJ-BFHQHQDPSA-N 0.000 description 1
- SOBIAADAMRHGKH-CIUDSAMLSA-N Ala-Leu-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O SOBIAADAMRHGKH-CIUDSAMLSA-N 0.000 description 1
- DEAGTWNKODHUIY-MRFFXTKBSA-N Ala-Tyr-Trp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O DEAGTWNKODHUIY-MRFFXTKBSA-N 0.000 description 1
- OMSKGWFGWCQFBD-KZVJFYERSA-N Ala-Val-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OMSKGWFGWCQFBD-KZVJFYERSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DAPLJWATMAXPPZ-CIUDSAMLSA-N Asn-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O DAPLJWATMAXPPZ-CIUDSAMLSA-N 0.000 description 1
- WONGRTVAMHFGBE-WDSKDSINSA-N Asn-Gly-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N WONGRTVAMHFGBE-WDSKDSINSA-N 0.000 description 1
- UGXYFDQFLVCDFC-CIUDSAMLSA-N Asn-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O UGXYFDQFLVCDFC-CIUDSAMLSA-N 0.000 description 1
- WLVLIYYBPPONRJ-GCJQMDKQSA-N Asn-Thr-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O WLVLIYYBPPONRJ-GCJQMDKQSA-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
- 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
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 101710112752 Cytotoxin Proteins 0.000 description 1
- WEAHRLBPCANXCN-UHFFFAOYSA-N Daunomycin Natural products CCC1(O)CC(OC2CC(N)C(O)C(C)O2)c3cc4C(=O)c5c(OC)cccc5C(=O)c4c(O)c3C1 WEAHRLBPCANXCN-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000011510 Elispot assay Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 229910052688 Gadolinium Inorganic materials 0.000 description 1
- YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 description 1
- MADFVRSKEIEZHZ-DCAQKATOSA-N Gln-Gln-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)N)N MADFVRSKEIEZHZ-DCAQKATOSA-N 0.000 description 1
- MLSKFHLRFVGNLL-WDCWCFNPSA-N Gln-Leu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MLSKFHLRFVGNLL-WDCWCFNPSA-N 0.000 description 1
- VPKBCVUDBNINAH-GARJFASQSA-N Glu-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)O)N)C(=O)O VPKBCVUDBNINAH-GARJFASQSA-N 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- LCNXZQROPKFGQK-WHFBIAKZSA-N Gly-Asp-Ser Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O LCNXZQROPKFGQK-WHFBIAKZSA-N 0.000 description 1
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 1
- DBUNZBWUWCIELX-JHEQGTHGSA-N Gly-Thr-Glu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DBUNZBWUWCIELX-JHEQGTHGSA-N 0.000 description 1
- IROABALAWGJQGM-OALUTQOASA-N Gly-Trp-Tyr Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)NC(=O)CN IROABALAWGJQGM-OALUTQOASA-N 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 101100337462 Homo sapiens GPC3 gene Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase 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
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 1
- LOLUPZNNADDTAA-AVGNSLFASA-N Leu-Gln-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LOLUPZNNADDTAA-AVGNSLFASA-N 0.000 description 1
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 1
- TUIOUEWKFFVNLH-DCAQKATOSA-N Leu-Val-Cys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(O)=O TUIOUEWKFFVNLH-DCAQKATOSA-N 0.000 description 1
- FMFNIDICDKEMOE-XUXIUFHCSA-N Leu-Val-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FMFNIDICDKEMOE-XUXIUFHCSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- UWKNTTJNVSYXPC-CIUDSAMLSA-N Lys-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN UWKNTTJNVSYXPC-CIUDSAMLSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 229930126263 Maytansine Natural products 0.000 description 1
- 206010027457 Metastases to liver Diseases 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 206010033128 Ovarian cancer Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 101150112800 PE35 gene Proteins 0.000 description 1
- 101150030083 PE38 gene Proteins 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- JARJPEMLQAWNBR-GUBZILKMSA-N Pro-Asp-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JARJPEMLQAWNBR-GUBZILKMSA-N 0.000 description 1
- PULPZRAHVFBVTO-DCAQKATOSA-N Pro-Glu-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O PULPZRAHVFBVTO-DCAQKATOSA-N 0.000 description 1
- VYWNORHENYEQDW-YUMQZZPRSA-N Pro-Gly-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 VYWNORHENYEQDW-YUMQZZPRSA-N 0.000 description 1
- DXTOOBDIIAJZBJ-BQBZGAKWSA-N Pro-Gly-Ser Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CO)C(O)=O DXTOOBDIIAJZBJ-BQBZGAKWSA-N 0.000 description 1
- FDMKYQQYJKYCLV-GUBZILKMSA-N Pro-Pro-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 FDMKYQQYJKYCLV-GUBZILKMSA-N 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100028964 Proteoglycan 3 Human genes 0.000 description 1
- 101710127914 Proteoglycan 3 Proteins 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- MWMKFWJYRRGXOR-ZLUOBGJFSA-N Ser-Ala-Asn Chemical compound N[C@H](C(=O)N[C@H](C(=O)N[C@H](C(=O)O)CC(N)=O)C)CO MWMKFWJYRRGXOR-ZLUOBGJFSA-N 0.000 description 1
- WTWGOQRNRFHFQD-JBDRJPRFSA-N Ser-Ala-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WTWGOQRNRFHFQD-JBDRJPRFSA-N 0.000 description 1
- MUARUIBTKQJKFY-WHFBIAKZSA-N Ser-Gly-Asp Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MUARUIBTKQJKFY-WHFBIAKZSA-N 0.000 description 1
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-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
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 1
- PURRNJBBXDDWLX-ZDLURKLDSA-N Ser-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N)O PURRNJBBXDDWLX-ZDLURKLDSA-N 0.000 description 1
- BDMWLJLPPUCLNV-XGEHTFHBSA-N Ser-Thr-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BDMWLJLPPUCLNV-XGEHTFHBSA-N 0.000 description 1
- QYBRQMLZDDJBSW-AVGNSLFASA-N Ser-Tyr-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O QYBRQMLZDDJBSW-AVGNSLFASA-N 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- MQCPGOZXFSYJPS-KZVJFYERSA-N Thr-Ala-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MQCPGOZXFSYJPS-KZVJFYERSA-N 0.000 description 1
- DIPIPFHFLPTCLK-LOKLDPHHSA-N Thr-Gln-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O DIPIPFHFLPTCLK-LOKLDPHHSA-N 0.000 description 1
- MXDOAJQRJBMGMO-FJXKBIBVSA-N Thr-Pro-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O MXDOAJQRJBMGMO-FJXKBIBVSA-N 0.000 description 1
- LECUEEHKUFYOOV-ZJDVBMNYSA-N Thr-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](N)[C@@H](C)O LECUEEHKUFYOOV-ZJDVBMNYSA-N 0.000 description 1
- YOPQYBJJNSIQGZ-JNPHEJMOSA-N Thr-Tyr-Tyr Chemical compound C([C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 YOPQYBJJNSIQGZ-JNPHEJMOSA-N 0.000 description 1
- PWONLXBUSVIZPH-RHYQMDGZSA-N Thr-Val-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O PWONLXBUSVIZPH-RHYQMDGZSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 206010052779 Transplant rejections Diseases 0.000 description 1
- OZUJUVFWMHTWCZ-HOCLYGCPSA-N Trp-Gly-His Chemical compound N[C@@H](Cc1c[nH]c2ccccc12)C(=O)NCC(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O OZUJUVFWMHTWCZ-HOCLYGCPSA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- CFSSLXZJEMERJY-NRPADANISA-N Val-Gln-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CFSSLXZJEMERJY-NRPADANISA-N 0.000 description 1
- DJQIUOKSNRBTSV-CYDGBPFRSA-N Val-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](C(C)C)N DJQIUOKSNRBTSV-CYDGBPFRSA-N 0.000 description 1
- GBIUHAYJGWVNLN-UHFFFAOYSA-N Val-Ser-Pro Natural products CC(C)C(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O GBIUHAYJGWVNLN-UHFFFAOYSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 1
- 210000001766 X chromosome Anatomy 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 108010004469 allophycocyanin Proteins 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 239000000611 antibody drug conjugate Substances 0.000 description 1
- 229940049595 antibody-drug conjugate Drugs 0.000 description 1
- 239000008365 aqueous carrier Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000011888 autopsy Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 230000022534 cell killing Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 239000002619 cytotoxin Substances 0.000 description 1
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 108010085059 glutamyl-arginyl-proline Proteins 0.000 description 1
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 1
- 108010010147 glycylglutamine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 235000014304 histidine Nutrition 0.000 description 1
- 150000002411 histidines Chemical class 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 208000002780 macular degeneration Diseases 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
- 238000005259 measurement Methods 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 238000004091 panning Methods 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 150000003905 phosphatidylinositols Chemical class 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 229910052761 rare earth metal Inorganic materials 0.000 description 1
- 150000002910 rare earth metals Chemical class 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 229910052707 ruthenium Inorganic materials 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- 239000001540 sodium lactate Substances 0.000 description 1
- 229940005581 sodium lactate Drugs 0.000 description 1
- 235000011088 sodium lactate Nutrition 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000036964 tight binding Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 108010044292 tryptophyltyrosine Proteins 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 229960003048 vinblastine Drugs 0.000 description 1
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
- C07K16/303—Liver or Pancreas
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
- A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
- A61K47/6817—Toxins
-
- 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
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/567—Framework region [FR]
Landscapes
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明提供了能结合GPC3蛋白的高亲和力的人源化单克隆抗体。所述人源化的单克隆抗体保留了与原始单克隆抗体相近的亲和力，具有更低的免疫原性，可用于开发活性更好的GPC3抗体药物、抗体‑药物缀合物或免疫毒素类药物。
Description
技术领域
本发明涉及抗体领域。具体涉及一类人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体。
背景技术
现今，抗体是医学和研究中广泛使用的物质。在医学中，发现它们在许多不同领域中的应用。例如，将抗体用作检测某些标记物的标记试剂，这使得可以诊断或预后疾病或测定特定的身体参数。例如,某些癌细胞表面的蛋白分子的检测。
此外，抗体还用作各种疾病的治疗和预防中的治疗剂，如癌症、心血管疾病、炎性疾病、黄斑变性、移植排斥、多发性硬化和病毒感染。在这些治疗中，抗体自身可以具有治疗活性，例如通过阻断受体或信使分子，由此抑制它们的疾病相关功能，或者通过恢复和激活患者的免疫系统的组成部分。或者，抗体可以与另一种具有治疗活性的物质结合。特别地，在癌症和感染的治疗中，所述另一种物质具有细胞杀灭活性，并且可以是，例如，放射性同位素或细胞毒素。在另一种应用中，通过将合适的抗体转移至患者的循环中，抗体可以用于被动地免疫患者。
目前单克隆抗体制备的常用有杂交瘤技术和噬菌体展示技术，能产生针对抗原的单个特异性表位的抗体。而这些抗体序列经常来源于哺乳动物，如鼠，兔。从进化的角度来看，哺乳动物的亲缘关系和人类很接近，很多蛋白都是同源的，所以在这些动物中产生相应的抗体。
鸡的亲缘关系与人更远，免疫鸡能产生覆盖更多表位的抗体。鸡抗体的框架区序列只有一种，也更容易构建噬菌体展示文库。
然而，在治疗应用中，这些单克隆抗体具有源自动物生物体并且其氨基酸序列与人抗体不同的问题。因此，人免疫系统将这些动物抗体识别为外来物质，并且将其从循环中快速清除。此外，可能导致全身性炎性效应。所以动物源的抗体需要进一步进行人源化改造。
抗体人源化的一种常用方法是互补决定区移植，其中将非人源抗体的CDRs移植到人源框架上。通常，选择与非人抗体框架区同源性最高的人构架作为CDR移植的受体。在单克隆抗体的人源化过程中，常常降低了工程化抗体的亲和力和特异性。而在实际应用中，抗原对所用抗体的紧密结合和特异性识别都是至关重要的。所以筛选最佳的框架序列既能让人源化程度更高，又能不丧失抗体的活性非常有意义。
GPC3(Glypican-3)是硫酸肝素蛋白聚糖的一员，通过糖基磷脂酰肌醇(glycosyl-phosphatidylinositol，GPI)锚定于细胞膜表面。人GPC3基因位于X染色体(Xp26)上并编码70kDa蛋白质，该蛋白质含有580个氨基酸，在Arg358和Ser359之间被弗林蛋白酶样转化酶内切切割，产生40kDa N末端亚基和30kDa C末端亚基，C末端亚基上还有两条硫酸乙酰肝素(heparan sulfate，HS)链。研究表明，GPC3在约72％肝细胞癌(hepatocellularcarcinoma，HCC)中的表达水平相比正常肝细胞、胆管癌和肝转移癌明显上调，但在正常成人的肝组织中不表达。此外，肝癌患者GPC3表达量高往往伴随着较差的预后，这些都表明GPC3在HCC中作为生物标志物的潜在作用。目前GPC3已被建议作为抗体和基于细胞的免疫疗法的靶标。
发明内容
本发明前期研究将GPC3-hFc蛋白免疫鸡后，利用噬菌体展示技术筛选得到抗体，然后通过噬菌体夹心ELISA将抗体按表位初步分类，利用抗体蛋白夹心ELISA的方法筛选到一系列能够高亲和力识别磷脂酰肌醇蛋白聚糖3不同表位的且具有良好的稳定性的抗体。本发明此基础上将这些GPC3高亲和力鸡抗体的框架区中大部分序列替换成人框架区后，人源化的抗体不仅免疫原性降低，而且保留有高亲和力的特性。
本发明具体技术方案如下：
本发明是基于具有与相应鸡抗体相似的抗原结合特性的人源化抗GPC3抗体的产生。
本发明提供了一类人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，所述人源化抗体包括重链可变区和轻链可变区，所述重链可变区的框架区：HFR1氨基酸序列如SEQ IDNO:3的1-25位所示，HFR2氨基酸序列如SEQ ID NO:3的35-49位或SEQ ID NO:19的35-49位所示，HFR3氨基酸序列如SEQ ID NO:3的67-98位，SEQ ID NO:15的67-98位，SEQ ID NO:17的67-98位中任一项所示，HFR4氨基酸序列如SEQ ID NO:3的116-126位所示，所述轻链可变区的框架区：LFR1氨基酸序列如SEQ ID NO:4的1-22位所示，LFR2氨基酸序列如SEQ ID NO:4的30-45位或SEQ ID NO:14的30-45位所示，LFR3氨基酸序列如SEQ ID NO:4的53-84位，SEQ ID NO:6的53-84位，SEQ ID NO:8的53-84位，SEQ ID NO:10的53-84位，SEQ ID NO:12的53-84位中任一项所示，LFR4氨基酸序列如SEQ ID NO:4的98-107位所示。
进一步的，所述的人源化抗体，优选自：
(1)所述人源化抗体的重链可变区具有SEQ ID NO:3的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:4的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(2)所述人源化抗体的重链可变区具有SEQ ID NO:5的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:6的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(3)所述人源化抗体的重链可变区具有SEQ ID NO:7的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:8的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(4)所述人源化抗体的重链可变区具有SEQ ID NO:9的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:10的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(5)所述人源化抗体的重链可变区具有SEQ ID NO:11的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:12的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(6)所述人源化抗体的重链可变区具有SEQ ID NO:13的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:14的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(7)所述人源化抗体的重链可变区具有SEQ ID NO:15的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:16的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(8)所述人源化抗体的重链可变区具有SEQ ID NO:17的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:18的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(9)所述人源化抗体的重链可变区具有SEQ ID NO:19的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:20的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
本发明一个具体的技术方案，所述磷脂酰肌醇蛋白聚糖3的人源化抗体，包括重链可变区和轻链可变区，
(1)所述抗体的重链可变区氨基酸序列如SEQ ID NO:3所示，轻链可变氨基酸序列如SEQ ID NO:4所示。
(2)所述抗体的重链可变区氨基酸序列如SEQ ID NO:5所示，轻链可变氨基酸序列如SEQ ID NO:6所示。
(3)所述抗体的重链可变区氨基酸序列如SEQ ID NO:7所示，轻链可变氨基酸序列如SEQ ID NO:8所示。
(4)所述抗体的重链可变区氨基酸序列如SEQ ID NO:9所示，轻链可变氨基酸序列如SEQ ID NO:10所示。
(5)所述抗体的重链可变区氨基酸序列如SEQ ID NO:11所示，轻链可变氨基酸序列如SEQ ID NO:12所示。
(6)所述抗体的重链可变区氨基酸序列如SEQ ID NO:13所示，轻链可变氨基酸序列如SEQ ID NO:14所示。
(7)所述抗体的重链可变区氨基酸序列如SEQ ID NO:15所示，轻链可变氨基酸序列如SEQ ID NO:16所示。
(8)所述抗体的重链可变区氨基酸序列如SEQ ID NO:17所示，轻链可变氨基酸序列如SEQ ID NO:18所示。
(9)所述抗体的重链可变区氨基酸序列如SEQ ID NO:19所示，轻链可变氨基酸序列如SEQ ID NO:20所示。
本发明一个具体的示例，所述人源化抗体的重链可变区具有SEQ ID NO:3的26-34位、50-66位和99-115位氨基酸残基所示的互补决定区；所述抗体的轻链可变区具有SEQ IDNO:4的23-29位、46-82位和85-97位氨基酸残基所示的互补决定区。
本发明所述的单克隆抗体可以为任何同种型。可以为例如IgM或IgG抗体，例如IgG1或IgG2。可特异性结合GPC3的抗体的种类可根据已知的方法彼此转换(例如，IgG可转换为IgM)。种类转换还可用于使一种IgG亚类转换成另一亚类，例如从IgG1转换成IgG2。
本发明所述的抗体可以为：
(1)Fab，包含抗体分子的单价抗原结合片段的片段，其可以通过用木瓜蛋白酶消化完整抗体以产生完整轻链和一条重链的一部分而产生；
(2)Fab'，可以通过用胃蛋白酶处理完整抗体、之后进行还原以产生完整轻链和重链的一部分而获得的抗体分子片段；每个抗体分子得到两个Fab'片段；
(3)(Fab')2，可以通过将完整的抗体用胃蛋白酶处理、但之后不进行还原而得到的抗体片段；F(ab')2是两个Fab'片段通过两个二硫键连接在一起的二聚体；
(4)Fv，含有表达为2条链的轻链可变区和重链可变区的基因程片段；
(5)单链抗体(例如scFv)，含有轻链可变区和重链可变区并通过合适的多肽接头将其连接为遗传上融合的单链分子的基因工程分子；
(6)单链抗体的二聚体(scFv2)，定义为scFv的二聚体(还被称为“微型抗体”)；
(7)VH单结构域抗体，由重链可变区组成的抗体片段。
本领域技术人员会了解，可以制备抗体的保守变体。可以在所述VH和/或VL区进行氨基酸置换(例如1个、2个、3个、4个或5个氨基酸置换)，置换后的VH和VL仍然保留结合GPC3的能力，或者对GPC3的结合能力更强。功能类似的氨基酸的保守置换是本领域普通技术人员所熟知的。以下六组是被认为是互为保守置换的氨基酸的实例：
1)丙氨酸(A)、丝氨酸(S)、苏氨酸(T)；
2)天冬氨酸(D)、谷氨酸(E)；
3)天冬酰胺(N)、谷氨酰胺(Q)；
4)精氨酸(R)、赖氨酸(K)；
5)异亮氨酸(I)、亮氨酸(L)、甲硫氨酸(M)、缬氨酸(V)；
6)苯丙氨酸(F)、酪氨酸(Y)、色氨酸(W)。
根据本发明的人源化抗体或其片段或衍生物优选用于医学中，特别是用于疾病的治疗、诊断、预后和监控中，特别是在此所述的疾病，优选癌症。
可将本发明所述的GPC3抗体缀合到效应分子上。效应分子包括但不限于毒素、药物或可检测的标记物。
本发明所述的药物是具有细胞毒性或抗肿瘤活性的物质，例如Monomethylauristatin E(MMAE)、Monomethylauristatin F(MMAF)、Pyrrolobenzodiazepine(PBD)dimer、N2'-deacetyl-N2'-(3-Mercapto-1-oxopropyl)-Maytansine(美登素DM1)、长春碱、道诺霉素等，又例如放射性试剂125I、32P、14C、3H和35S等。
本发明所述的毒素为具有细胞毒性或抗肿瘤活性的毒蛋白，可以与本发明所述抗体缀合形成免疫毒素，包括但不限于假单胞菌外毒素、蓖麻毒蛋白、相思豆毒蛋白、白喉毒素和其亚基以及肉毒杆菌毒素A-F，以及这些毒素的截短突变体和点突变体。这些毒素可商业购买获得(例如，Sigma Chemical Company，St.Louis，MO)。所述毒素还包括上述毒素的变体(例如参见美国专利No.5,079,163和4,689,401)。在一个实施方案中，所述毒素为假单胞菌外毒素(PE)(美国专利No.5,602,095)。所述“假单胞菌外毒素”包括其天然序列、所述天然序列的细胞毒性片段、以及天然序列或其细胞毒性片段的保守修饰的变体。这些修饰包括但不限于除去结构域Ia、结构域Ib、II和III中的多个氨基酸缺失、单个或多个氨基酸置换以及在羧基端加入一个或多个序列(例如，参见，Siegall et al.，J.Biol.Chem.264：14256-14261，1989)。假单胞菌外毒素的细胞毒性片段包括PE24、PE40、PE38和PE35等。
本发明所述可检测的标记物是可以被同位素分析仪、酶标仪、生物发光检测仪、化学发光检测仪、电化学发光检测仪、荧光分析仪器检测，或裸眼可视化的物质，这些物质包括但不限于放射性同位素(如3H、14C、15N、35S、90Y、、99Tc、111In、125I、131I)、可用于检测的酶类(如辣根过氧化物酶、β-半乳糖苷酶、碱性磷酸酶、葡萄糖氧化酶等)、荧光蛋白(如绿色荧光蛋白(GFP)、黄色荧光蛋白(YFP)、别藻蓝蛋白APC、藻红蛋白PE)、生物发光标记物(如萤光素酶)、荧光化合物(如荧光素、异硫氰酸荧光素、罗丹明、5-二甲基氨基-1-萘磺酰氯、藻红蛋白、荧光染料Cy3、Cy5、稀土无机发光材料、量子点等)、生物素、磁性试剂(例如钆)、电化学发光试剂(如三联吡啶钌)、胶体金。
本发明另一目的在于提供一种多核苷酸，编码本发明所述的抗体，重组蛋白或者免疫缀合物。本发明另一目的在于提供一种载体，含有本发明所述的多核苷酸。所述的载体包括：细菌质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒、或其他载体。
本发明提供了包含根据本发明的核酸或根据本发明的表达盒或载体的宿主细胞。根据本发明的宿主细胞可以是任何宿主细胞。可以是分离的细胞或包含在组织中的细胞。优选地，宿主细胞是培养的细胞，特别是初级细胞或已建立的细胞系的细胞，优选源自肿瘤的人细胞。
本发明另一目的在于提供一种药物组合物，包括本发明所述的抗体、重组蛋白、免疫缀合物、多核苷酸、载体或遗传工程化的宿主细胞中的一种或几种。所述药物组合物还包括药学上可接受的载体。所述抗体、重组蛋白、免疫缀合物、多核苷酸、载体或遗传工程化的宿主细胞可溶于水性载体，例如缓冲盐水等。还可以含有接近生理条件所需要的可药用辅料，例如pH调节剂和缓冲试剂等，乙酸钠、氯化钠、氯化钾、氯化钙和乳酸钠等。
本发明另一目的在于提供本发明所述的抗体、重组蛋白、免疫缀合物、多核苷酸、载体或遗传工程化的宿主细胞在制备自身免疫疾病、病毒感染或癌症的治疗药物或诊断试剂中的应用。所述癌症为肝癌、胃癌、结直肠癌、肺癌或卵巢癌，或表达GPC3的任何其他类型的癌症。
本发明公开的单克隆抗体还可用于制备嵌合抗原受体(CAR；也称为嵌合T细胞受体、人造T细胞受体或嵌合免疫受体)或双特异性抗体。
本发明所提供的抗体和组合物可用于多种目的，例如用于肿瘤的分子诊断，在肝癌和其它肿瘤患者样品中确认是否表达GPC3。所述样品可以为任何样品，包括但不限于来自活组织检查、尸体解剖和病理标本的组织。生物样品还包括组织的切片，例如为组织学目的获取的冷冻切片。生物样品还包括体液，例如血液、血清、血浆、痰、脊髓液或尿。生物样品一般获自哺乳动物，包括人，非人灵长类，小鼠等。
本发明优点：本发明提供了GPC3鸡单克隆抗体的人源化策略。以GPC3鸡单克隆抗体A5为例，本实施例以A5抗体为例，将其重链和轻链可变区(VH，SEQ ID NO:21和VL，SEQ IDNO:22)用连接肽G4S连接，形成scFv的结构。基于scFv的结构，根据其整体序列相似性和CDR环分类，从人种系抗体文库中选择最相关的框架区：重链可变区的框架区1、2和3选自人种系VH基因3-23*02。重链可变区的框架区4选自人种系基因J3*01。轻链可变区的框架区1、2和3源自人种系VL基因3-25*02。轻链可变区的框架区4选自人种系基因J7*01，并进行优化，得到的人源化抗体具有理想的亲和力，并且免疫原性更低。可用于开发活性更好的GPC3抗体药物、抗体-药物缀合物或免疫毒素类药物。
附图说明
图1FACS法检测本发明实施例1制备的抗体A5与GPC3阴性/阳性细胞系的结合活性。
图2本发明实施例4人源化鸡抗体重链可变区的框架区和互补决定区的序列比对差异。
图3本发明实施例4人源化鸡抗体轻链可变区的框架区和互补决定区的序列比对差异。
图4.SDS-PAGE检测HuA5.2抗体及8个突变体蛋白的纯度。
图5.ELISA检测HuA5.2抗体及8个突变体与抗原GPC3结合的亲和力。
图6.SDS-PAGE检测真核表达系统纯化的HuA5.2抗体的纯度
图7.ELISA检测HuA5.2和A5抗体与抗原结合的亲和力
图8.ELIspot检测HuA5.2和A5抗体的免疫原性
具体实施方式
下面结合具体实施例并参照数据进一步详细描述本发明，应理解，这些实施例只是为了举例说明本发明，而非以任何方式限制本发明的范围。在本发明中使用的术语，除非另有说明，一般具有本领域普通技术人员通常理解的含义。在以下实施例中，未详细描述的各种过程和方法是本领域中公知的常规方法。
实施例1结合GPC3的鸡单克隆抗体A5的制备
以GPC3-hFc蛋白(Sino Biological，Cat:10088-H02H2)免疫4月龄鸡，每次免疫300μg，每次间隔2周，免疫4次之后采集鸡的脾脏，提取脾脏中的总RNA，逆转录为cDNA，作为建立噬菌体抗体文库的模板，并构建鸡抗体噬菌体文库(库容量2.7×109)。利用GPC3蛋白作为抗原，对噬菌体文库进行4次淘选后，随机挑取300个克隆进行测序，比对序列后发现A5是其中出现频次最高的抗体。
将A5的scFv与hFc融合，构建表达载体pPBSPS-A5-hFc，并在293F细胞中表达。表达产物用protein A层析柱(GE healthcare)进行纯化。利用纯化的抗体和夹心ELISA方法对GPC3蛋白进行检测。
A5的VH如SEQ ID NO:21所示，VL如SEQ ID NO:22所示，其中CDR及HFR区序列分别如表1所示。
表1 GPC3鸡单克隆抗体的可变区氨基酸序列
实施例2结合GPC3的鸡单克隆抗体的体外表征
2.SPR测定实施例1制得的抗体与GPC3的结合动力学与亲和力
将GPC3-his蛋白通过标准的胺偶联固定在羧甲基传感器芯片上(S系列传感器芯片CM5)。洗涤芯片以获取稳定的基线，随后以30μL/分钟的流速，将不同浓度的抗体分析物以及运行缓冲液注入芯片，样品结合时间为180秒，随后的解离时间为600秒。使用ProteOn软件将结合和解离曲线拟合至1:1Langmiur结合模型。结果显示A5的亲和力常数Kd值为1.82E-11，表明其与GPC3的亲和力非常高。
3.抗体稳定性的检测
利用Nano Temper公司的Prometheus NT.48测定A5的稳定性：将抗体浓度稀释成50ug/ml，然后上样，1.5℃/min的速度，40min钟内温度从25℃上升到95℃。最后得出抗体的Tm值。结果显示A5的Tm值为69.1℃，说明A5的热稳定性好。
实施例3FACS检测本发明所述抗体与GPC3阳性细胞系的结合
将A431，G1(超表达GPC3的A431细胞系)(Phung Yet al.Mabs 2012；4:592-599)和肝癌细胞系HepG2、Hep3B、HuH-7以贴壁方式培养，所用培养基为DMEM培养基(Invitrogen，Carlsbad，CA)并添加10％胎牛血清(HyClone，Logan，UT)、1％L-谷氨酰胺和1％青霉素-链霉素(Invitrogen，Carlsbad，CA)。收获细胞后，将实施例1制得的抗体分别与A431，G1，HepG2，Hep3B，Huh7细胞进行结合，加入APC标记的羊抗人的二抗，检测结合至细胞表面上的抗体。结果如图1所示，实施例1制得的抗体能很强的结合G1细胞和HepG2，Hep3B，Huh7细胞，但不结合GPC3阴性的A431细胞，说明实施例1制得的抗体能特异性地识别并结合细胞表面的GPC3蛋白。
实施例4抗GPC3抗体的鸡重链和轻链可变区的人源化
本实施例以A5抗体为例，将其重链和轻链可变区(VH，SEQ ID NO:21和VL，SEQ IDNO:22)用连接肽G4S连接，形成scFv的结构。基于scFv的结构，根据其整体序列相似性和CDR环分类，从人种系抗体文库中选择最相关的框架区：重链可变区的框架区1、2和3选自人种系VH基因3-23*02。重链可变区的框架区4选自人种系基因J3*01。轻链可变区的框架区1、2和3源自人种系VL基因3-25*02。轻链可变区的框架区4选自人种系基因J7*01。
人源化方案一：将鸡重链可变区和轻链可变区的框架区1、2、3、4全部替换为人源化框架区，所述抗体的重链可变区具有SEQ ID NO:1的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:2的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区。将此方案得到的抗体序列命名为HuA5.1。
人源化方案二：参照文献资料(Leeying Wu,J Immunol 2012；188:322-333)，将鸡重链可变区和轻链可变区的框架区1、2、3、4中突变率高的位置，替换为人源化框架区对应位置的氨基酸，所述抗体的重链可变区具有SEQ ID NO:3的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:4的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；。将此方案得到的抗体序列命名为HuA5.2。
人源化方案三：在人源化方案二的基础上，对鸡重链可变区和轻链可变区的框架区1、2、3、4中突变率低，也就是比较保守的鸡序列逐个将替换成人的序列。参照Kabat和IMGT编号规则，分别将HuA5.2重链可变区35位G突变成S，67位A突变成F，78位V突变成L。分别将HuA5.2轻链可变区36位F变成Y，57位D变成G，60位S变成D,66位A变成S，69位S变成N。将此方案得到的抗体序列分别命名为H-G35S,H-A67F,H-V78L,L-F36Y,L-D57G,L-S60D,L-A66S,L-S69N。
共得到10条不同序列的人源化抗体。所述抗体重链和轻链序列如表2所示，其可变区的框架区如表3所示。所述抗体重链可变区的框架区和互补决定区的序列比对差异如图2所示。所述抗体轻链可变区的框架区和互补决定区的序列比对差异如图3所示。
表2. 10个人源化抗体的可变区氨基酸序列
表3. 10个人源化抗体的FR氨基酸序列(根据Kabat和IMTG数据库)
实施例5表达实施例4中的人源化抗体并检测其纯度
委托金斯瑞公司合成实施例1中抗体对应的的核酸片段，然后将抗体的序列C末端额外加上6个组氨酸(6x His)，构建相应抗体的pET28a表达载体后，转化BL21大肠杆菌，然后将菌株接种到2L的2YT培养基中，37℃培养4-5h后OD600达到0.9，加入1mM IPTG诱导，30℃的条件下表达10h。收集菌体，高压破碎菌体后离心，过滤处理上清液，然后通过镍柱进行纯化。其中，HuA5.1抗体无法表达，其余抗体均能表达纯化。结果表明，将框架区全部替换成人源框架后，抗体的构象发生很大变化，最终导致抗体无法纯化。最后将其余纯化后的蛋白经过SDS-PAGE检测其纯度。实施例4制得的抗体的大小约为50Kd，如图4所示，条带大小正确，其中4个抗体纯度比较高，剩余5个抗体纯度一般，但均满足后续实验的要求。
实施例6ELISA检测实施例5制备得到的人源化抗体结合GPC3的活性
将GPC3蛋白包被在ELISA板底，然后将实施例4中的抗体蛋白分别以20μg/ml开始1：3梯度稀释后加入孔内，然后加入HRP-mouse anti His抗体，最后加入底物显色。如图5所示，所有抗体HuA5.2、H-G35S,H-A67F,H-V78L,L-F36Y,L-D57G,L-S60D,L-A66S,L-S69N均能显著地与GPC3抗体进行结合。根据ELISA地结果计算抗体地亲和力常数Kd值，如表4所示，其中HuA5.2的亲和力最高，Kd值达到了0.02051。结果表明HuA5.2在人源化后，亲和力损失最少。
表4实施例5中的抗体结合GPC3蛋白的亲和力EC50值
实施例7 293细胞表达HuA5.2蛋白并检测纯度
考虑后续要检测HuA5.2抗体的免疫原性，在哺乳动物细胞表达系统中表达HuA5.2，避免大肠杆菌表达系统额外引入的免疫原。在HuA5.2的序列的C端加上额外的hFc标签，构建哺乳动物细胞表达载体，然后将构建成功的载体瞬转293F细胞，悬浮培养7天后收集细胞上清，过滤后用ProteinA层析柱纯化。利用SDS-PAGE检测其纯度。HuA5.2抗体的大小约为100Kd，还原后大小约为50Kd，如图6所示，蛋白的条带大小均正确。
实施例8比较HuA5.2和A5结合GPC3的亲和力差异
将GPC3抗原蛋白(Sino Biological，Cat:10088-H02H2)包被在ELISA板底，然后将HuA5.2和鸡抗A5分别以首孔20nM的浓度以1：3梯度稀释后加入孔内，然后加入HRP-羊抗人抗体，最后显色。如图7所示，HuA5.2的EC50值为0.08628，而A5的EC50值为0.0961，说明HuA5.2与GPC3蛋白的结合能力相比A5来说并没有降低，甚至亲和力更高。
实施例9ELIspot检测HuA5.2抗体的免疫原性
Human IFN-γprecoated ELISPOT kit购自于达科为有限公司，货号为Cat#.211005(strips)，实验方法参见试剂盒的说明书。简单来说，就是将抗体与PNMC在ELISPOT板上共孵育，然后通过板底已经固定的IFN-γ抗体捕获PBMC分泌的IFN-γ，然后显色在板底形成斑点，通过统计斑点数目的多少来判断HuA5.2的免疫原性的高低。斑点数越多，说明PBMC分泌的IFN-γ越多，对所检蛋白的免疫反应越强，也就是免疫原性越大，反之亦然。如图8所示，HuA5.2的斑点数明显少于A5，这表明HuA5.2刺激T细胞产生的IFN-γ更少，免疫原性更低。
序列表
<110> 华中农业大学
<120> 人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用
<160> 22
<170> SIPOSequenceListing 1.0
<210> 1
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 2
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln
1 5 10 15
Thr Ala Arg Ile Thr Cys Ser Gly Asp Gly Ser Tyr Ala Tyr Trp Tyr
20 25 30
Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Asp Asn Asp
35 40 45
Glu Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser Ser Ser Gly
50 55 60
Thr Thr Val Thr Leu Thr Ile Ser Gly Val Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Gln Leu Thr Val Leu
100 105
<210> 3
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 4
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 5
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 6
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 7
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 8
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ser Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 9
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 10
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Asp Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 11
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 12
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 13
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 14
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Tyr Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 15
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 16
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 17
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 18
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 19
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 20
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Gln Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Ala Ser Gly
50 55 60
Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu Ala
65 70 75 80
Asp Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 21
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly
1 5 10 15
Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Asn Arg Tyr
20 25 30
Cys Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Asp Ser Asp Ser Gly Gly Thr Asp Tyr Gly Ala Ala Val
50 55 60
Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg
65 70 75 80
Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Thr Tyr Tyr Cys
85 90 95
Ala Lys Ser Ala Tyr Gly Gly Trp Cys Gly Ser Arg Val Ala Pro Trp
100 105 110
Ile Asp Ala Trp Gly His Gly Thr Glu Val Ile Val Ser Ser
115 120 125
<210> 22
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Gln Ala Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu
1 5 10 15
Thr Val Lys Ile Thr Cys Ser Gly Gly Gly Ser Tyr Gly Trp Phe Gln
20 25 30
Gln Lys Ser Pro Gly Ser Ala Pro Val Thr Val Ile Tyr Asp Asn Asp
35 40 45
Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Leu Ser Gly
50 55 60
Ser Thr Gly Thr Leu Thr Ile Thr Gly Val Arg Ala Glu Asp Glu Ala
65 70 75 80
Val Tyr Tyr Cys Gly Ser Ser Glu Asn Ser Tyr Val Gly His Val Ala
85 90 95
Ile Phe Gly Ala Gly Thr Thr Leu Thr Val Leu
100 105
Claims (13)
1.人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，其特征在于所述抗体包括重链可变区和轻链可变区，所述重链可变区的框架区：HFR1氨基酸序列如SEQ ID NO:3的1-25位所示，HFR2氨基酸序列如SEQ ID NO:3的35-49位或SEQ ID NO:19的35-49位所示，HFR3氨基酸序列如SEQ ID NO:3的67-98位，SEQ ID NO:15的67-98位，SEQ ID NO:17的67-98位中任一项所示，HFR4氨基酸序列如SEQ ID NO:3的116-126位所示，所述轻链可变区的框架区：LFR1氨基酸序列如SEQ ID NO:4的1-22位所示，LFR2氨基酸序列如SEQ ID NO:4的30-45位或SEQID NO:14的30-45位所示，LFR3氨基酸序列如SEQ ID NO:4的53-84位，SEQ ID NO:6的53-84位，SEQ ID NO:8的53-84位，SEQ ID NO:10的53-84位，SEQ ID NO:12的53-84位中任一项所示，LFR4氨基酸序列如SEQ ID NO:4的98-107位所示。
2.如权利要求1所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，其特征在于：
(1)所述抗体的重链可变区具有SEQ ID NO:3的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:4的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(2)所述抗体的重链可变区具有SEQ ID NO:5的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:6的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(3)所述抗体的重链可变区具有SEQ ID NO:7的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:8的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(4)所述抗体的重链可变区具有SEQ ID NO:9的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:10的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(5)所述抗体的重链可变区具有SEQ ID NO:11的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:12的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(6)所述抗体的重链可变区具有SEQ ID NO:13的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:14的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(7)所述抗体的重链可变区具有SEQ ID NO:15的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:16的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(8)所述抗体的重链可变区具有SEQ ID NO:17的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:18的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区；
(9)所述抗体的重链可变区具有SEQ ID NO:19的1-25位、35-49位、67-98位和116-126位氨基酸残基所示的框架区；所述抗体的轻链可变区具有SEQ ID NO:20的1-22位、30-45位、53-84和98-107位氨基酸残基所示的框架区。
3.如权利要求1所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，其特征在于：
1)所述抗体的重链可变区氨基酸序列如SEQ ID NO:3所示，轻链可变氨基酸序列如SEQID NO:4所示。
2)所述抗体的重链可变区氨基酸序列如SEQ ID NO:5所示，轻链可变氨基酸序列如SEQID NO:6所示。
3)所述抗体的重链可变区氨基酸序列如SEQ ID NO:7所示，轻链可变氨基酸序列如SEQID NO:8所示。
4)所述抗体的重链可变区氨基酸序列如SEQ ID NO:9所示，轻链可变氨基酸序列如SEQID NO:10所示。
5)所述抗体的重链可变区氨基酸序列如SEQ ID NO:11所示，轻链可变氨基酸序列如SEQ ID NO:12所示。
6)所述抗体的重链可变区氨基酸序列如SEQ ID NO:13所示，轻链可变氨基酸序列如SEQ ID NO:14所示。
7)所述抗体的重链可变区氨基酸序列如SEQ ID NO:15所示，轻链可变氨基酸序列如SEQ ID NO:16所示。
8)所述抗体的重链可变区氨基酸序列如SEQ ID NO:17所示，轻链可变氨基酸序列如SEQ ID NO:18所示。
9)所述抗体的重链可变区氨基酸序列如SEQ ID NO:19所示，轻链可变氨基酸序列如SEQ ID NO:20所示。
4.如权利要求1或2所述的单克隆抗体，其特征在于所述抗体的重链可变区具有SEQ IDNO:3的26-34位、50-66位和99-115位氨基酸残基所示的互补决定区；所述抗体的轻链可变区具有SEQ ID NO:4的23-29位、46-82位和85-97位氨基酸残基所示的互补决定区。
5.如权利要求1-4任一项所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，其特征在于还包含协助表达和/或纯化的标签序列。
6.一种免疫缀合物，其特征在于包含权利要求1-5任一项所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体和效应分子。
7.根据权利要求6所述的免疫缀合物，其特征在于所述效应分子是毒素、药物或可检测的标记物。
8.根据权利要求7所述的免疫缀合物，其特征在于所述毒素是具有细胞毒性或抗肿瘤活性的蛋白，包括相思豆毒蛋白或其变体、蓖麻毒蛋白或其变体、假单胞菌外毒素或其变体、白喉毒素或其变体、肉毒杆菌毒素或其变体；所述药物是具有细胞毒性或抗肿瘤活性的化合物，包括Monomethyl auristatin E，Monomethylauristatin F，Pyrrolobenzodiazepine dimer，N2'-deacetyl-N2'-(3-Mercapto-1-oxopropyl)-Maytansine；所述可检测的标记物是可以被同位素分析仪、酶标仪、生物发光检测仪、化学发光检测仪、电化学发光检测仪、荧光分析仪器检测，或裸眼可视化的物质。
9.一种多核苷酸，其特征在于其编码根据权利要求1-5任一项所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，或权利要求6-8任一项所述的免疫缀合物。
10.一种载体，其特征在于含有权利要求9所述的多核苷酸。
11.一种遗传工程化的宿主细胞，其特征在于含有权利要求10所述的载体，或基因组中整合有权利要求8所述的多核苷酸。
12.一种药物组合物，其特征在于包括根据权利要求1-5任一项所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，权利要求6-8任一项所述的免疫缀合物，权利要求9所述的多核苷酸，权利要求10所述的载体或者权利要求11所述的遗传工程化的宿主细胞中的一种或几种。
13.根据权利要求1-5任一项所述的人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体，或权利要求6-8任一项所述的免疫缀合物、权利要求9所述的多核苷酸，权利要求10所述的载体或者权利要求11所述的遗传工程化的宿主细胞在制备自身免疫疾病、病毒感染或癌症的治疗药物或诊断试剂中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011158008.2A CN114478779B (zh) | 2020-10-26 | 2020-10-26 | 人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011158008.2A CN114478779B (zh) | 2020-10-26 | 2020-10-26 | 人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114478779A true CN114478779A (zh) | 2022-05-13 |
| CN114478779B CN114478779B (zh) | 2024-10-11 |
Family
ID=81470803
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011158008.2A Active CN114478779B (zh) | 2020-10-26 | 2020-10-26 | 人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114478779B (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103596985A (zh) * | 2011-04-19 | 2014-02-19 | 美国政府(由卫生和人类服务部的部长所代表) | 对磷脂酰肌醇蛋白聚糖3特异的人单克隆抗体及其用途 |
| US20150147330A1 (en) * | 2012-06-01 | 2015-05-28 | The United States of America,as represented by the Secretary,Department of Health and Human Service | High-affinity monoclonal antibodies to glypican-3 and use thereof |
| CN110627904A (zh) * | 2019-10-31 | 2019-12-31 | 南京蓝盾生物科技有限公司 | 抗人gpc3单克隆抗体 |
-
2020
- 2020-10-26 CN CN202011158008.2A patent/CN114478779B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103596985A (zh) * | 2011-04-19 | 2014-02-19 | 美国政府(由卫生和人类服务部的部长所代表) | 对磷脂酰肌醇蛋白聚糖3特异的人单克隆抗体及其用途 |
| US20150147330A1 (en) * | 2012-06-01 | 2015-05-28 | The United States of America,as represented by the Secretary,Department of Health and Human Service | High-affinity monoclonal antibodies to glypican-3 and use thereof |
| CN110627904A (zh) * | 2019-10-31 | 2019-12-31 | 南京蓝盾生物科技有限公司 | 抗人gpc3单克隆抗体 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114478779B (zh) | 2024-10-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2018177393A1 (zh) | B7-h3抗体、其抗原结合片段及其医药用途 | |
| WO2019091449A1 (zh) | Cd96抗体、其抗原结合片段及医药用途 | |
| US20220089709A1 (en) | Protein binding to fibronectin b domain | |
| CN109988240B (zh) | 抗gpc-3抗体及其用途 | |
| CN113330036B (zh) | 结合pd-l1和ox40的双特异性抗体 | |
| CN113307870B (zh) | 抗il5纳米抗体及其应用 | |
| US20180002439A1 (en) | Anti-mesothelin antibodies and uses thereof | |
| CN113227148B (zh) | 抗gpc3抗体、其抗原结合片段及其医药用途 | |
| CN112250762B (zh) | 一种抗hiv的广谱中和抗体 | |
| AU2020212534A1 (en) | High affinity monoclonal antibodies targeting glypican-1 and methods of use | |
| WO2012071216A2 (en) | Antibodies for tumor gangliosides | |
| CN115298216A (zh) | 抗体或其抗原结合片段、其制备方法及医药用途 | |
| CN115505043A (zh) | 特异性结合糖基化ceacam5的抗体 | |
| WO2023207475A1 (zh) | 靶向gpc3的单克隆抗体和包含此抗体的抗体药物缀合物 | |
| US20240218055A1 (en) | Lassa virus-specific nanobodies and methods of their use | |
| CN113880948B (zh) | 抗ca724抗体或其抗原结合片段及其制备方法和应用 | |
| CN113461820B (zh) | 抗cd3人源化抗体 | |
| US20230002503A1 (en) | Nano-antibody targeting caix antigen and application thereof | |
| CN114478779B (zh) | 人源化的磷脂酰肌醇蛋白聚糖3的单克隆抗体及其应用 | |
| CN111018984B (zh) | 抗ck8单克隆抗体及其应用 | |
| CN115843256A (zh) | 抗erbb3抗体或其抗原结合片段及其医药用途 | |
| CN114763383A (zh) | 靶向人bcma的单克隆抗体及其应用 | |
| CN114349864B (zh) | 抗前列腺酸性磷酸酶抗体及其用途 | |
| WO2023035226A1 (zh) | 抗ang2抗体及其制备方法和应用 | |
| US12122843B2 (en) | High affinity monoclonal antibodies targeting glypican-1 and methods of use |
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 | ||
| GR01 | Patent grant |