CN109929036A - 一种表位特异的抗体筛选方法及所筛选到的抗体 - Google Patents
一种表位特异的抗体筛选方法及所筛选到的抗体 Download PDFInfo
- Publication number
- CN109929036A CN109929036A CN201711375292.7A CN201711375292A CN109929036A CN 109929036 A CN109929036 A CN 109929036A CN 201711375292 A CN201711375292 A CN 201711375292A CN 109929036 A CN109929036 A CN 109929036A
- Authority
- CN
- China
- Prior art keywords
- seq
- antibody
- ser
- screening
- epitope
- 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
- 238000012216 screening Methods 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 27
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 30
- 239000003814 drug Substances 0.000 claims abstract description 12
- 229940079593 drug Drugs 0.000 claims abstract description 11
- 201000011510 cancer Diseases 0.000 claims abstract description 6
- 208000026278 immune system disease Diseases 0.000 claims abstract 2
- 210000004027 cell Anatomy 0.000 claims description 63
- 150000001413 amino acids Chemical class 0.000 claims description 43
- 235000001014 amino acid Nutrition 0.000 claims description 23
- 102000001189 Cyclic Peptides Human genes 0.000 claims description 15
- 108010069514 Cyclic Peptides Proteins 0.000 claims description 15
- 230000014509 gene expression Effects 0.000 claims description 14
- 235000018417 cysteine Nutrition 0.000 claims description 11
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 11
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 8
- 239000002773 nucleotide Substances 0.000 claims description 7
- 125000003729 nucleotide group Chemical group 0.000 claims description 7
- 239000004471 Glycine Substances 0.000 claims description 5
- 239000000427 antigen Substances 0.000 claims description 4
- 102000036639 antigens Human genes 0.000 claims description 4
- 108091007433 antigens Proteins 0.000 claims description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 4
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 3
- 230000008878 coupling Effects 0.000 claims description 3
- 238000010168 coupling process Methods 0.000 claims description 3
- 238000005859 coupling reaction Methods 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 2
- 101710160107 Outer membrane protein A Proteins 0.000 claims description 2
- 230000003321 amplification Effects 0.000 claims description 2
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 2
- 241001515965 unidentified phage Species 0.000 claims 3
- 239000002253 acid Substances 0.000 claims 2
- 230000000295 complement effect Effects 0.000 claims 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims 1
- 108010008038 Synthetic Vaccines Proteins 0.000 claims 1
- 239000003937 drug carrier Substances 0.000 claims 1
- 230000009870 specific binding Effects 0.000 claims 1
- 229940124597 therapeutic agent Drugs 0.000 claims 1
- 230000027455 binding Effects 0.000 abstract description 29
- 230000001225 therapeutic effect Effects 0.000 abstract description 7
- 239000003446 ligand Substances 0.000 abstract description 6
- 238000010171 animal model Methods 0.000 abstract description 4
- 238000005516 engineering process Methods 0.000 abstract description 4
- 238000002823 phage display Methods 0.000 abstract description 4
- 238000009169 immunotherapy Methods 0.000 abstract 1
- 230000008685 targeting Effects 0.000 abstract 1
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 45
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 44
- 229960002621 pembrolizumab Drugs 0.000 description 27
- 101000611936 Homo sapiens Programmed cell death protein 1 Proteins 0.000 description 23
- 102000048362 human PDCD1 Human genes 0.000 description 22
- 229960003301 nivolumab Drugs 0.000 description 19
- 239000011324 bead Substances 0.000 description 17
- 108090000623 proteins and genes Proteins 0.000 description 15
- 239000002953 phosphate buffered saline Substances 0.000 description 13
- 108010074708 B7-H1 Antigen Proteins 0.000 description 10
- 102000008096 B7-H1 Antigen Human genes 0.000 description 10
- 206010028980 Neoplasm Diseases 0.000 description 10
- 210000001744 T-lymphocyte Anatomy 0.000 description 10
- 238000002965 ELISA Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 8
- 102000004196 processed proteins & peptides Human genes 0.000 description 8
- 235000018102 proteins Nutrition 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 238000010276 construction Methods 0.000 description 7
- 102220330185 rs1364834319 Human genes 0.000 description 7
- 230000000890 antigenic effect Effects 0.000 description 6
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 5
- RXUOAOOZIWABBW-XGEHTFHBSA-N Ser-Thr-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RXUOAOOZIWABBW-XGEHTFHBSA-N 0.000 description 5
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 108010037850 glycylvaline Proteins 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 5
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 5
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 5
- 108091026890 Coding region Proteins 0.000 description 4
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 4
- PVMPDMIKUVNOBD-CIUDSAMLSA-N Leu-Asp-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O PVMPDMIKUVNOBD-CIUDSAMLSA-N 0.000 description 4
- 206010027480 Metastatic malignant melanoma Diseases 0.000 description 4
- 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 4
- 230000008827 biological function Effects 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 230000006957 competitive inhibition Effects 0.000 description 4
- 239000013604 expression vector Substances 0.000 description 4
- 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 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 108010017391 lysylvaline Proteins 0.000 description 4
- 208000021039 metastatic melanoma Diseases 0.000 description 4
- 230000035772 mutation Effects 0.000 description 4
- 238000013207 serial dilution Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- ARHJJAAWNWOACN-FXQIFTODSA-N Ala-Ser-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O ARHJJAAWNWOACN-FXQIFTODSA-N 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 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 3
- GBYYQVBXFVDJPJ-WLTAIBSBSA-N Gly-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)CN)O GBYYQVBXFVDJPJ-WLTAIBSBSA-N 0.000 description 3
- 241000880493 Leptailurus serval Species 0.000 description 3
- PXHCFKXNSBJSTQ-KKUMJFAQSA-N Lys-Asn-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N)O PXHCFKXNSBJSTQ-KKUMJFAQSA-N 0.000 description 3
- 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 3
- GNRMAQSIROFNMI-IXOXFDKPSA-N Phe-Thr-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O GNRMAQSIROFNMI-IXOXFDKPSA-N 0.000 description 3
- 108010076504 Protein Sorting Signals Proteins 0.000 description 3
- 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 3
- JQAWYCUUFIMTHE-WLTAIBSBSA-N Thr-Gly-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JQAWYCUUFIMTHE-WLTAIBSBSA-N 0.000 description 3
- YYLHVUCSTXXKBS-IHRRRGAJSA-N Tyr-Pro-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O YYLHVUCSTXXKBS-IHRRRGAJSA-N 0.000 description 3
- 108010062796 arginyllysine Proteins 0.000 description 3
- 108010068265 aspartyltyrosine Proteins 0.000 description 3
- 125000004122 cyclic group Chemical group 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 108010015792 glycyllysine Proteins 0.000 description 3
- 210000000987 immune system Anatomy 0.000 description 3
- 230000001394 metastastic effect Effects 0.000 description 3
- 206010061289 metastatic neoplasm Diseases 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 108010077037 tyrosyl-tyrosyl-phenylalanine Proteins 0.000 description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- 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 2
- JBGSZRYCXBPWGX-BQBZGAKWSA-N Ala-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N JBGSZRYCXBPWGX-BQBZGAKWSA-N 0.000 description 2
- BTRULDJUUVGRNE-DCAQKATOSA-N Ala-Pro-Lys Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O BTRULDJUUVGRNE-DCAQKATOSA-N 0.000 description 2
- CREYEAPXISDKSB-FQPOAREZSA-N Ala-Thr-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CREYEAPXISDKSB-FQPOAREZSA-N 0.000 description 2
- 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 2
- 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 2
- 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 2
- 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 2
- XYBJLTKSGFBLCS-QXEWZRGKSA-N Asp-Arg-Val Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC(O)=O XYBJLTKSGFBLCS-QXEWZRGKSA-N 0.000 description 2
- QNFRBNZGVVKBNJ-PEFMBERDSA-N Asp-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N QNFRBNZGVVKBNJ-PEFMBERDSA-N 0.000 description 2
- 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 2
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 2
- 229940045513 CTLA4 antagonist Drugs 0.000 description 2
- NIXHTNJAGGFBAW-CIUDSAMLSA-N Cys-Lys-Ser Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N NIXHTNJAGGFBAW-CIUDSAMLSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 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 2
- JXFLPKSDLDEOQK-JHEQGTHGSA-N Gln-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O JXFLPKSDLDEOQK-JHEQGTHGSA-N 0.000 description 2
- 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 2
- PAQUJCSYVIBPLC-AVGNSLFASA-N Glu-Asp-Phe Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 PAQUJCSYVIBPLC-AVGNSLFASA-N 0.000 description 2
- RFTVTKBHDXCEEX-WDSKDSINSA-N Glu-Ser-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RFTVTKBHDXCEEX-WDSKDSINSA-N 0.000 description 2
- 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 2
- 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 2
- HAOUOFNNJJLVNS-BQBZGAKWSA-N Gly-Pro-Ser Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O HAOUOFNNJJLVNS-BQBZGAKWSA-N 0.000 description 2
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-N 0.000 description 2
- 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 2
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 2
- 101100519206 Homo sapiens PDCD1 gene Proteins 0.000 description 2
- PXKACEXYLPBMAD-JBDRJPRFSA-N Ile-Ser-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PXKACEXYLPBMAD-JBDRJPRFSA-N 0.000 description 2
- 102000037982 Immune checkpoint proteins Human genes 0.000 description 2
- 108091008036 Immune checkpoint proteins Proteins 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 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 2
- HXWALXSAVBLTPK-NUTKFTJISA-N Leu-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(C)C)N HXWALXSAVBLTPK-NUTKFTJISA-N 0.000 description 2
- 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 2
- CQGSYZCULZMEDE-SRVKXCTJSA-N Leu-Gln-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O CQGSYZCULZMEDE-SRVKXCTJSA-N 0.000 description 2
- CQGSYZCULZMEDE-UHFFFAOYSA-N Leu-Gln-Pro Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)N1CCCC1C(O)=O CQGSYZCULZMEDE-UHFFFAOYSA-N 0.000 description 2
- 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 2
- FAELBUXXFQLUAX-AJNGGQMLSA-N Leu-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C FAELBUXXFQLUAX-AJNGGQMLSA-N 0.000 description 2
- 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 2
- SUYRAPCRSCCPAK-VFAJRCTISA-N Leu-Trp-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SUYRAPCRSCCPAK-VFAJRCTISA-N 0.000 description 2
- 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 2
- YQFZRHYZLARWDY-IHRRRGAJSA-N Leu-Val-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN YQFZRHYZLARWDY-IHRRRGAJSA-N 0.000 description 2
- FLCMXEFCTLXBTL-DCAQKATOSA-N Lys-Asp-Arg Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N FLCMXEFCTLXBTL-DCAQKATOSA-N 0.000 description 2
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 2
- SPSSJSICDYYTQN-HJGDQZAQSA-N Met-Thr-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O SPSSJSICDYYTQN-HJGDQZAQSA-N 0.000 description 2
- 101150087384 PDCD1 gene Proteins 0.000 description 2
- YYKZDTVQHTUKDW-RYUDHWBXSA-N Phe-Gly-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N YYKZDTVQHTUKDW-RYUDHWBXSA-N 0.000 description 2
- 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 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
- KLYYKKGCPOGDPE-OEAJRASXSA-N Phe-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O KLYYKKGCPOGDPE-OEAJRASXSA-N 0.000 description 2
- 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 2
- 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 2
- FMDHKPRACUXATF-ACZMJKKPSA-N Ser-Gln-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O FMDHKPRACUXATF-ACZMJKKPSA-N 0.000 description 2
- 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 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
- 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 2
- 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 2
- AZWNCEBQZXELEZ-FXQIFTODSA-N Ser-Pro-Ser Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O AZWNCEBQZXELEZ-FXQIFTODSA-N 0.000 description 2
- PQEQXWRVHQAAKS-SRVKXCTJSA-N Ser-Tyr-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CO)N)CC1=CC=C(O)C=C1 PQEQXWRVHQAAKS-SRVKXCTJSA-N 0.000 description 2
- UBTNVMGPMYDYIU-HJPIBITLSA-N Ser-Tyr-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UBTNVMGPMYDYIU-HJPIBITLSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- LKEKWDJCJSPXNI-IRIUXVKKSA-N Thr-Glu-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 LKEKWDJCJSPXNI-IRIUXVKKSA-N 0.000 description 2
- GXUWHVZYDAHFSV-FLBSBUHZSA-N Thr-Ile-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GXUWHVZYDAHFSV-FLBSBUHZSA-N 0.000 description 2
- RQLNEFOBQAVGSY-WDSOQIARSA-N Trp-Met-Leu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O RQLNEFOBQAVGSY-WDSOQIARSA-N 0.000 description 2
- 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 2
- FFCRCJZJARTYCG-KKUMJFAQSA-N Tyr-Cys-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCCN)C(=O)O)N)O FFCRCJZJARTYCG-KKUMJFAQSA-N 0.000 description 2
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 2
- LMKKMCGTDANZTR-BZSNNMDCSA-N Tyr-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LMKKMCGTDANZTR-BZSNNMDCSA-N 0.000 description 2
- ZYVAAYAOTVJBSS-GMVOTWDCSA-N Tyr-Trp-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(O)=O ZYVAAYAOTVJBSS-GMVOTWDCSA-N 0.000 description 2
- NUQZCPSZHGIYTA-HKUYNNGSSA-N Tyr-Trp-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N NUQZCPSZHGIYTA-HKUYNNGSSA-N 0.000 description 2
- WKWJJQZZZBBWKV-JYJNAYRXSA-N Val-Arg-Tyr Chemical compound NC(N)=NCCC[C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 WKWJJQZZZBBWKV-JYJNAYRXSA-N 0.000 description 2
- VCAWFLIWYNMHQP-UKJIMTQDSA-N Val-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N VCAWFLIWYNMHQP-UKJIMTQDSA-N 0.000 description 2
- 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 2
- 108010081404 acein-2 Proteins 0.000 description 2
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 2
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 2
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 108010013835 arginine glutamate Proteins 0.000 description 2
- 108010008355 arginyl-glutamine Proteins 0.000 description 2
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 230000002860 competitive effect Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 2
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 2
- 238000013537 high throughput screening Methods 0.000 description 2
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 2
- 238000000464 low-speed centrifugation Methods 0.000 description 2
- 108010064235 lysylglycine Proteins 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 210000001948 pro-b lymphocyte Anatomy 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 235000013930 proline Nutrition 0.000 description 2
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 2
- 108010070643 prolylglutamic acid Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 108010044292 tryptophyltyrosine Proteins 0.000 description 2
- 108010035534 tyrosyl-leucyl-alanine Proteins 0.000 description 2
- 108010003137 tyrosyltyrosine Proteins 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- PIPTUBPKYFRLCP-NHCYSSNCSA-N Ala-Ala-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 PIPTUBPKYFRLCP-NHCYSSNCSA-N 0.000 description 1
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 1
- KUDREHRZRIVKHS-UWJYBYFXSA-N Ala-Asp-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KUDREHRZRIVKHS-UWJYBYFXSA-N 0.000 description 1
- SFNFGFDRYJKZKN-XQXXSGGOSA-N Ala-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](C)N)O SFNFGFDRYJKZKN-XQXXSGGOSA-N 0.000 description 1
- 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 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- 108010011667 Ala-Phe-Ala Proteins 0.000 description 1
- DYJJJCHDHLEFDW-FXQIFTODSA-N Ala-Pro-Cys Chemical compound C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CS)C(=O)O)N DYJJJCHDHLEFDW-FXQIFTODSA-N 0.000 description 1
- IORKCNUBHNIMKY-CIUDSAMLSA-N Ala-Pro-Glu Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O IORKCNUBHNIMKY-CIUDSAMLSA-N 0.000 description 1
- YJHKTAMKPGFJCT-NRPADANISA-N Ala-Val-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O YJHKTAMKPGFJCT-NRPADANISA-N 0.000 description 1
- PNQWAUXQDBIJDY-GUBZILKMSA-N Arg-Glu-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PNQWAUXQDBIJDY-GUBZILKMSA-N 0.000 description 1
- XRNXPIGJPQHCPC-RCWTZXSCSA-N Arg-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)O)C(O)=O XRNXPIGJPQHCPC-RCWTZXSCSA-N 0.000 description 1
- ZUVMUOOHJYNJPP-XIRDDKMYSA-N Arg-Trp-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZUVMUOOHJYNJPP-XIRDDKMYSA-N 0.000 description 1
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 1
- XYOVHPDDWCEUDY-CIUDSAMLSA-N Asn-Ala-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O XYOVHPDDWCEUDY-CIUDSAMLSA-N 0.000 description 1
- GXMSVVBIAMWMKO-BQBZGAKWSA-N Asn-Arg-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CCCN=C(N)N GXMSVVBIAMWMKO-BQBZGAKWSA-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
- LGCVSPFCFXWUEY-IHPCNDPISA-N Asn-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)[C@H](CC(=O)N)N LGCVSPFCFXWUEY-IHPCNDPISA-N 0.000 description 1
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 1
- UZFHNLYQWMGUHU-DCAQKATOSA-N Asp-Lys-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UZFHNLYQWMGUHU-DCAQKATOSA-N 0.000 description 1
- GKWFMNNNYZHJHV-SRVKXCTJSA-N Asp-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC(O)=O GKWFMNNNYZHJHV-SRVKXCTJSA-N 0.000 description 1
- DPNWSMBUYCLEDG-CIUDSAMLSA-N Asp-Lys-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O DPNWSMBUYCLEDG-CIUDSAMLSA-N 0.000 description 1
- SQIARYGNVQWOSB-BZSNNMDCSA-N Asp-Tyr-Phe 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=CC=C1)C(O)=O SQIARYGNVQWOSB-BZSNNMDCSA-N 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 101100421200 Caenorhabditis elegans sep-1 gene Proteins 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102000000844 Cell Surface Receptors Human genes 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- VIRYODQIWJNWNU-NRPADANISA-N Cys-Glu-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CS)N VIRYODQIWJNWNU-NRPADANISA-N 0.000 description 1
- KSMSFCBQBQPFAD-GUBZILKMSA-N Cys-Pro-Pro Chemical compound SC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 KSMSFCBQBQPFAD-GUBZILKMSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102000001301 EGF receptor Human genes 0.000 description 1
- 108060006698 EGF receptor Proteins 0.000 description 1
- 101000867232 Escherichia coli Heat-stable enterotoxin II Proteins 0.000 description 1
- DHNWZLGBTPUTQQ-QEJZJMRPSA-N Gln-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N DHNWZLGBTPUTQQ-QEJZJMRPSA-N 0.000 description 1
- IULKWYSYZSURJK-AVGNSLFASA-N Gln-Leu-Lys Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O IULKWYSYZSURJK-AVGNSLFASA-N 0.000 description 1
- AQPZYBSRDRZBAG-AVGNSLFASA-N Gln-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N AQPZYBSRDRZBAG-AVGNSLFASA-N 0.000 description 1
- LPIKVBWNNVFHCQ-GUBZILKMSA-N Gln-Ser-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O LPIKVBWNNVFHCQ-GUBZILKMSA-N 0.000 description 1
- FITIQFSXXBKFFM-NRPADANISA-N Gln-Val-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O FITIQFSXXBKFFM-NRPADANISA-N 0.000 description 1
- JJKKWYQVHRUSDG-GUBZILKMSA-N Glu-Ala-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O JJKKWYQVHRUSDG-GUBZILKMSA-N 0.000 description 1
- GLWXKFRTOHKGIT-ACZMJKKPSA-N Glu-Asn-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O GLWXKFRTOHKGIT-ACZMJKKPSA-N 0.000 description 1
- CKOFNWCLWRYUHK-XHNCKOQMSA-N Glu-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)O)N)C(=O)O CKOFNWCLWRYUHK-XHNCKOQMSA-N 0.000 description 1
- CLROYXHHUZELFX-FXQIFTODSA-N Glu-Gln-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O CLROYXHHUZELFX-FXQIFTODSA-N 0.000 description 1
- KASDBWKLWJKTLJ-GUBZILKMSA-N Glu-Glu-Met Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O KASDBWKLWJKTLJ-GUBZILKMSA-N 0.000 description 1
- QDMVXRNLOPTPIE-WDCWCFNPSA-N Glu-Lys-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QDMVXRNLOPTPIE-WDCWCFNPSA-N 0.000 description 1
- VSVZIEVNUYDAFR-YUMQZZPRSA-N Gly-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN VSVZIEVNUYDAFR-YUMQZZPRSA-N 0.000 description 1
- GRIRDMVMJJDZKV-RCOVLWMOSA-N Gly-Asn-Val Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O GRIRDMVMJJDZKV-RCOVLWMOSA-N 0.000 description 1
- UUYBFNKHOCJCHT-VHSXEESVSA-N Gly-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN UUYBFNKHOCJCHT-VHSXEESVSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- GJHWILMUOANXTG-WPRPVWTQSA-N Gly-Val-Arg Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GJHWILMUOANXTG-WPRPVWTQSA-N 0.000 description 1
- MAABHGXCIBEYQR-XVYDVKMFSA-N His-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N MAABHGXCIBEYQR-XVYDVKMFSA-N 0.000 description 1
- HIAHVKLTHNOENC-HGNGGELXSA-N His-Glu-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HIAHVKLTHNOENC-HGNGGELXSA-N 0.000 description 1
- 101001117317 Homo sapiens Programmed cell death 1 ligand 1 Proteins 0.000 description 1
- QQFSKBMCAKWHLG-UHFFFAOYSA-N Ile-Phe-Pro-Pro Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(NC(=O)C(N)C(C)CC)CC1=CC=CC=C1 QQFSKBMCAKWHLG-UHFFFAOYSA-N 0.000 description 1
- JHNJNTMTZHEDLJ-NAKRPEOUSA-N Ile-Ser-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JHNJNTMTZHEDLJ-NAKRPEOUSA-N 0.000 description 1
- VGSPNSSCMOHRRR-BJDJZHNGSA-N Ile-Ser-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N VGSPNSSCMOHRRR-BJDJZHNGSA-N 0.000 description 1
- 108010065920 Insulin Lispro Proteins 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
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 1
- DBVWMYGBVFCRBE-CIUDSAMLSA-N Leu-Asn-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DBVWMYGBVFCRBE-CIUDSAMLSA-N 0.000 description 1
- OIARJGNVARWKFP-YUMQZZPRSA-N Leu-Asn-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIARJGNVARWKFP-YUMQZZPRSA-N 0.000 description 1
- APFJUBGRZGMQFF-QWRGUYRKSA-N Leu-Gly-Lys Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN APFJUBGRZGMQFF-QWRGUYRKSA-N 0.000 description 1
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- BTNXKBVLWJBTNR-SRVKXCTJSA-N Leu-His-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O BTNXKBVLWJBTNR-SRVKXCTJSA-N 0.000 description 1
- AMSSKPUHBUQBOQ-SRVKXCTJSA-N Leu-Ser-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N AMSSKPUHBUQBOQ-SRVKXCTJSA-N 0.000 description 1
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 1
- LINKCQUOMUDLKN-KATARQTJSA-N Leu-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(C)C)N)O LINKCQUOMUDLKN-KATARQTJSA-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
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- MWVUEPNEPWMFBD-SRVKXCTJSA-N Lys-Cys-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CCCCN MWVUEPNEPWMFBD-SRVKXCTJSA-N 0.000 description 1
- NNCDAORZCMPZPX-GUBZILKMSA-N Lys-Gln-Ser Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N NNCDAORZCMPZPX-GUBZILKMSA-N 0.000 description 1
- ODUQLUADRKMHOZ-JYJNAYRXSA-N Lys-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCCCN)N)O ODUQLUADRKMHOZ-JYJNAYRXSA-N 0.000 description 1
- XNKDCYABMBBEKN-IUCAKERBSA-N Lys-Gly-Gln Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O XNKDCYABMBBEKN-IUCAKERBSA-N 0.000 description 1
- FGMHXLULNHTPID-KKUMJFAQSA-N Lys-His-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CN=CN1 FGMHXLULNHTPID-KKUMJFAQSA-N 0.000 description 1
- YTJFXEDRUOQGSP-DCAQKATOSA-N Lys-Pro-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O YTJFXEDRUOQGSP-DCAQKATOSA-N 0.000 description 1
- IOQWIOPSKJOEKI-SRVKXCTJSA-N Lys-Ser-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IOQWIOPSKJOEKI-SRVKXCTJSA-N 0.000 description 1
- SQXZLVXQXWILKW-KKUMJFAQSA-N Lys-Ser-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQXZLVXQXWILKW-KKUMJFAQSA-N 0.000 description 1
- DIBZLYZXTSVGLN-CIUDSAMLSA-N Lys-Ser-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O DIBZLYZXTSVGLN-CIUDSAMLSA-N 0.000 description 1
- YKBSXQFZWFXFIB-VOAKCMCISA-N Lys-Thr-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCCN)C(O)=O YKBSXQFZWFXFIB-VOAKCMCISA-N 0.000 description 1
- YFQSSOAGMZGXFT-MEYUZBJRSA-N Lys-Thr-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YFQSSOAGMZGXFT-MEYUZBJRSA-N 0.000 description 1
- RQILLQOQXLZTCK-KBPBESRZSA-N Lys-Tyr-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O RQILLQOQXLZTCK-KBPBESRZSA-N 0.000 description 1
- QLFAPXUXEBAWEK-NHCYSSNCSA-N Lys-Val-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QLFAPXUXEBAWEK-NHCYSSNCSA-N 0.000 description 1
- 108091092878 Microsatellite Proteins 0.000 description 1
- 101100407308 Mus musculus Pdcd1lg2 gene Proteins 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
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- SMFGCTXUBWEPKM-KBPBESRZSA-N Phe-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 SMFGCTXUBWEPKM-KBPBESRZSA-N 0.000 description 1
- JDMKQHSHKJHAHR-UHFFFAOYSA-N Phe-Phe-Leu-Tyr Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)CC1=CC=CC=C1 JDMKQHSHKJHAHR-UHFFFAOYSA-N 0.000 description 1
- JLLJTMHNXQTMCK-UBHSHLNASA-N Phe-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=CC=C1 JLLJTMHNXQTMCK-UBHSHLNASA-N 0.000 description 1
- IIEOLPMQYRBZCN-SRVKXCTJSA-N Phe-Ser-Cys Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O IIEOLPMQYRBZCN-SRVKXCTJSA-N 0.000 description 1
- SJRQWEDYTKYHHL-SLFFLAALSA-N Phe-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC3=CC=CC=C3)N)C(=O)O SJRQWEDYTKYHHL-SLFFLAALSA-N 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- IHCXPSYCHXFXKT-DCAQKATOSA-N Pro-Arg-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O IHCXPSYCHXFXKT-DCAQKATOSA-N 0.000 description 1
- TUYWCHPXKQTISF-LPEHRKFASA-N Pro-Cys-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CS)C(=O)N2CCC[C@@H]2C(=O)O TUYWCHPXKQTISF-LPEHRKFASA-N 0.000 description 1
- UAYHMOIGIQZLFR-NHCYSSNCSA-N Pro-Gln-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O UAYHMOIGIQZLFR-NHCYSSNCSA-N 0.000 description 1
- LGSANCBHSMDFDY-GARJFASQSA-N Pro-Glu-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)O)C(=O)N2CCC[C@@H]2C(=O)O LGSANCBHSMDFDY-GARJFASQSA-N 0.000 description 1
- ZLXKLMHAMDENIO-DCAQKATOSA-N Pro-Lys-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLXKLMHAMDENIO-DCAQKATOSA-N 0.000 description 1
- HWLKHNDRXWTFTN-GUBZILKMSA-N Pro-Pro-Cys Chemical compound C1C[C@H](NC1)C(=O)N2CCC[C@H]2C(=O)N[C@@H](CS)C(=O)O HWLKHNDRXWTFTN-GUBZILKMSA-N 0.000 description 1
- PCWLNNZTBJTZRN-AVGNSLFASA-N Pro-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 PCWLNNZTBJTZRN-AVGNSLFASA-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
- MKGIILKDUGDRRO-FXQIFTODSA-N Pro-Ser-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 MKGIILKDUGDRRO-FXQIFTODSA-N 0.000 description 1
- YDTUEBLEAVANFH-RCWTZXSCSA-N Pro-Val-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 YDTUEBLEAVANFH-RCWTZXSCSA-N 0.000 description 1
- 108700030875 Programmed Cell Death 1 Ligand 2 Proteins 0.000 description 1
- 102100024213 Programmed cell death 1 ligand 2 Human genes 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 238000011579 SCID mouse model Methods 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- QVOGDCQNGLBNCR-FXQIFTODSA-N Ser-Arg-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O QVOGDCQNGLBNCR-FXQIFTODSA-N 0.000 description 1
- VGNYHOBZJKWRGI-CIUDSAMLSA-N Ser-Asn-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO VGNYHOBZJKWRGI-CIUDSAMLSA-N 0.000 description 1
- FTVRVZNYIYWJGB-ACZMJKKPSA-N Ser-Asp-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FTVRVZNYIYWJGB-ACZMJKKPSA-N 0.000 description 1
- 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 1
- ZOHGLPQGEHSLPD-FXQIFTODSA-N Ser-Gln-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZOHGLPQGEHSLPD-FXQIFTODSA-N 0.000 description 1
- BPMRXBZYPGYPJN-WHFBIAKZSA-N Ser-Gly-Asn Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O BPMRXBZYPGYPJN-WHFBIAKZSA-N 0.000 description 1
- GZFAWAQTEYDKII-YUMQZZPRSA-N Ser-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO GZFAWAQTEYDKII-YUMQZZPRSA-N 0.000 description 1
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 1
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 1
- HHJFMHQYEAAOBM-ZLUOBGJFSA-N Ser-Ser-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O HHJFMHQYEAAOBM-ZLUOBGJFSA-N 0.000 description 1
- JCLAFVNDBJMLBC-JBDRJPRFSA-N Ser-Ser-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JCLAFVNDBJMLBC-JBDRJPRFSA-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
- 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 1
- PCJLFYBAQZQOFE-KATARQTJSA-N Ser-Thr-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N)O PCJLFYBAQZQOFE-KATARQTJSA-N 0.000 description 1
- ZKOKTQPHFMRSJP-YJRXYDGGSA-N Ser-Thr-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZKOKTQPHFMRSJP-YJRXYDGGSA-N 0.000 description 1
- AXKJPUBALUNJEO-UBHSHLNASA-N Ser-Trp-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(O)=O AXKJPUBALUNJEO-UBHSHLNASA-N 0.000 description 1
- RCOUFINCYASMDN-GUBZILKMSA-N Ser-Val-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(O)=O RCOUFINCYASMDN-GUBZILKMSA-N 0.000 description 1
- HNDMFDBQXYZSRM-IHRRRGAJSA-N Ser-Val-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HNDMFDBQXYZSRM-IHRRRGAJSA-N 0.000 description 1
- SIEBDTCABMZCLF-XGEHTFHBSA-N Ser-Val-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SIEBDTCABMZCLF-XGEHTFHBSA-N 0.000 description 1
- 208000000102 Squamous Cell Carcinoma of Head and Neck Diseases 0.000 description 1
- UKBSDLHIKIXJKH-HJGDQZAQSA-N Thr-Arg-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O UKBSDLHIKIXJKH-HJGDQZAQSA-N 0.000 description 1
- ODSAPYVQSLDRSR-LKXGYXEUSA-N Thr-Cys-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O ODSAPYVQSLDRSR-LKXGYXEUSA-N 0.000 description 1
- DSLHSTIUAPKERR-XGEHTFHBSA-N Thr-Cys-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O DSLHSTIUAPKERR-XGEHTFHBSA-N 0.000 description 1
- CYVQBKQYQGEELV-NKIYYHGXSA-N Thr-His-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O CYVQBKQYQGEELV-NKIYYHGXSA-N 0.000 description 1
- FIFDDJFLNVAVMS-RHYQMDGZSA-N Thr-Leu-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(O)=O FIFDDJFLNVAVMS-RHYQMDGZSA-N 0.000 description 1
- 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 1
- BDGBHYCAZJPLHX-HJGDQZAQSA-N Thr-Lys-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BDGBHYCAZJPLHX-HJGDQZAQSA-N 0.000 description 1
- DXPURPNJDFCKKO-RHYQMDGZSA-N Thr-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DXPURPNJDFCKKO-RHYQMDGZSA-N 0.000 description 1
- XKWABWFMQXMUMT-HJGDQZAQSA-N Thr-Pro-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O XKWABWFMQXMUMT-HJGDQZAQSA-N 0.000 description 1
- MROIJTGJGIDEEJ-RCWTZXSCSA-N Thr-Pro-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 MROIJTGJGIDEEJ-RCWTZXSCSA-N 0.000 description 1
- XHWCDRUPDNSDAZ-XKBZYTNZSA-N Thr-Ser-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N)O XHWCDRUPDNSDAZ-XKBZYTNZSA-N 0.000 description 1
- SGAOHNPSEPVAFP-ZDLURKLDSA-N Thr-Ser-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SGAOHNPSEPVAFP-ZDLURKLDSA-N 0.000 description 1
- CYCGARJWIQWPQM-YJRXYDGGSA-N Thr-Tyr-Ser Chemical compound C[C@@H](O)[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CO)C([O-])=O)CC1=CC=C(O)C=C1 CYCGARJWIQWPQM-YJRXYDGGSA-N 0.000 description 1
- AVYVKJMBNLPWRX-WFBYXXMGSA-N Trp-Ala-Ser Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 AVYVKJMBNLPWRX-WFBYXXMGSA-N 0.000 description 1
- UDCHKDYNMRJYMI-QEJZJMRPSA-N Trp-Glu-Ser Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O UDCHKDYNMRJYMI-QEJZJMRPSA-N 0.000 description 1
- AYHSJESDFKREAR-KKUMJFAQSA-N Tyr-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AYHSJESDFKREAR-KKUMJFAQSA-N 0.000 description 1
- SINRIKQYQJRGDQ-MEYUZBJRSA-N Tyr-Lys-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 SINRIKQYQJRGDQ-MEYUZBJRSA-N 0.000 description 1
- AUZADXNWQMBZOO-JYJNAYRXSA-N Tyr-Pro-Arg Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C1=CC=C(O)C=C1 AUZADXNWQMBZOO-JYJNAYRXSA-N 0.000 description 1
- RWOKVQUCENPXGE-IHRRRGAJSA-N Tyr-Ser-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RWOKVQUCENPXGE-IHRRRGAJSA-N 0.000 description 1
- MQGGXGKQSVEQHR-KKUMJFAQSA-N Tyr-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 MQGGXGKQSVEQHR-KKUMJFAQSA-N 0.000 description 1
- RIVVDNTUSRVTQT-IRIUXVKKSA-N Tyr-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O RIVVDNTUSRVTQT-IRIUXVKKSA-N 0.000 description 1
- PWKMJDQXKCENMF-MEYUZBJRSA-N Tyr-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O PWKMJDQXKCENMF-MEYUZBJRSA-N 0.000 description 1
- TYGHOWWWMTWVKM-HJOGWXRNSA-N Tyr-Tyr-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 TYGHOWWWMTWVKM-HJOGWXRNSA-N 0.000 description 1
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 1
- ZQGPWORGSNRQLN-NHCYSSNCSA-N Val-Asp-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N ZQGPWORGSNRQLN-NHCYSSNCSA-N 0.000 description 1
- LHADRQBREKTRLR-DCAQKATOSA-N Val-Cys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](C(C)C)N LHADRQBREKTRLR-DCAQKATOSA-N 0.000 description 1
- OXVPMZVGCAPFIG-BQFCYCMXSA-N Val-Gln-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N OXVPMZVGCAPFIG-BQFCYCMXSA-N 0.000 description 1
- OQWNEUXPKHIEJO-NRPADANISA-N Val-Glu-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N OQWNEUXPKHIEJO-NRPADANISA-N 0.000 description 1
- UEHRGZCNLSWGHK-DLOVCJGASA-N Val-Glu-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O UEHRGZCNLSWGHK-DLOVCJGASA-N 0.000 description 1
- ZIGZPYJXIWLQFC-QTKMDUPCSA-N Val-His-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](C(C)C)N)O ZIGZPYJXIWLQFC-QTKMDUPCSA-N 0.000 description 1
- HGJRMXOWUWVUOA-GVXVVHGQSA-N Val-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N HGJRMXOWUWVUOA-GVXVVHGQSA-N 0.000 description 1
- XTDDIVQWDXMRJL-IHRRRGAJSA-N Val-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N XTDDIVQWDXMRJL-IHRRRGAJSA-N 0.000 description 1
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 1
- HJSLDXZAZGFPDK-ULQDDVLXSA-N Val-Phe-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](C(C)C)N HJSLDXZAZGFPDK-ULQDDVLXSA-N 0.000 description 1
- VCIYTVOBLZHFSC-XHSDSOJGSA-N Val-Phe-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N VCIYTVOBLZHFSC-XHSDSOJGSA-N 0.000 description 1
- RYHUIHUOYRNNIE-NRPADANISA-N Val-Ser-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N RYHUIHUOYRNNIE-NRPADANISA-N 0.000 description 1
- QPJSIBAOZBVELU-BPNCWPANSA-N Val-Tyr-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N QPJSIBAOZBVELU-BPNCWPANSA-N 0.000 description 1
- ZLNYBMWGPOKSLW-LSJOCFKGSA-N Val-Val-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLNYBMWGPOKSLW-LSJOCFKGSA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 108010084455 Zeocin Proteins 0.000 description 1
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000001188 anti-phage Effects 0.000 description 1
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 1
- 230000005975 antitumor immune response Effects 0.000 description 1
- 238000011398 antitumor immunotherapy Methods 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010009111 arginyl-glycyl-glutamic acid Proteins 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit 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
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000001531 bladder carcinoma Diseases 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229960005395 cetuximab Drugs 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 208000013056 classic Hodgkin lymphoma Diseases 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 231100000655 enterotoxin Toxicity 0.000 description 1
- 230000017188 evasion or tolerance of host immune response Effects 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
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 1
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 1
- 108010050475 glycyl-leucyl-tyrosine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 201000000459 head and neck squamous cell carcinoma Diseases 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 102000048776 human CD274 Human genes 0.000 description 1
- 230000008629 immune suppression Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004073 interleukin-2 production Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229960005386 ipilimumab Drugs 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 231100000150 mutagenicity / genotoxicity testing Toxicity 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 238000011022 operating instruction Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 108010024654 phenylalanyl-prolyl-alanine Proteins 0.000 description 1
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 125000001500 prolyl group Chemical class [H]N1C([H])(C(=O)[*])C([H])([H])C([H])([H])C1([H])[H] 0.000 description 1
- 108010090894 prolylleucine Proteins 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 210000003289 regulatory T cell Anatomy 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000013391 scatchard analysis Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 238000009097 single-agent therapy Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- VUFNRPJNRFOTGK-UHFFFAOYSA-M sodium;1-[4-[(2,5-dioxopyrrol-1-yl)methyl]cyclohexanecarbonyl]oxy-2,5-dioxopyrrolidine-3-sulfonate Chemical compound [Na+].O=C1C(S(=O)(=O)[O-])CC(=O)N1OC(=O)C1CCC(CN2C(C=CC2=O)=O)CC1 VUFNRPJNRFOTGK-UHFFFAOYSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000000153 supplemental effect Effects 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 206010044412 transitional cell carcinoma Diseases 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 108010080629 tryptophan-leucine Proteins 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 108010079202 tyrosyl-alanyl-cysteine Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010071635 tyrosyl-prolyl-arginine Proteins 0.000 description 1
- 208000010570 urinary bladder carcinoma Diseases 0.000 description 1
- 208000023747 urothelial carcinoma Diseases 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 108010027345 wheylin-1 peptide Proteins 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
一种表位特异的抗体筛选方法，以解决抗体库筛选技术中具有治疗作用的功能性抗体筛选效率低的问题，包括用功能表位肽筛选和活细胞筛选的步骤；还提供了用该方法筛选噬菌体展示抗体库所获得的一种全人源抗PD‑1单克隆抗体，针对特定的功能性表位，能有效阻断PD‑1与其配体的结合，并具有亲和力高、EC50低的特点，能在低剂量下对动物模型产生治疗作用。本发明大大提高了功能性抗体的筛选效率，并为恶性肿瘤、免疫疾病患者提供了一种潜在的更安全经济的免疫治疗药物。
Description
技术领域
本发明属于生物医药领域，具体涉及一种表位特异的抗体筛选方法及所筛选到的抗体。
背景技术
癌细胞的“免疫逃逸”被认为是肿瘤发生、发展和抗药的主要机制。肿瘤可通过直接或间接抑制T细胞的信号来保护自己躲过免疫系统的清除。肿瘤的免疫检查点疗法（immune chenkpoint therapy）是一类通过共抑制或共刺激信号等一系列途径调节T细胞活性来提高抗肿瘤免疫应答的治疗方法。
PD-1（程序性细胞死亡蛋白1，英文全称Programmed Cell Death Protein 1，也称为CD279）是表达于活化的T细胞和前B细胞（pro-B cells）上的一种细胞表面受体，是一个免疫检查点，对免疫系统有负调控作用，通过抑制T细胞炎性活动促进自身耐受，更具体而言，PD-1通过促进淋巴结中抗原特异性T细胞的凋亡和降低调控T细胞（Treg，也叫抑制性T细胞）的凋亡的双重机制防止自身免疫的发生。
PD-1由PDCD1基因编码，全长288个氨基酸，其中N端前20个氨基酸为信号肽、在成熟后切除，第21-170个氨基酸为胞外段，第171-191个氨基酸为跨膜区，第192-288个氨基酸为胞内尾。
PD-1可与2种配体结合——B7家族成员PD-L1和PD-L2。在小鼠上，PD-L1广泛表达于心、肺、胸腺、脾、肾等多种器官上，以及几乎所有的小鼠肿瘤细胞系，并在多种人类肿瘤细胞上高表达；而PD-L2的表达较为局限，主要是树突状细胞和少数肿瘤细胞系。因此，PD-1及其配体PD-L1成为抗肿瘤免疫治疗的靶点。
目前已有2个抗PD-1靶向治疗药物被批准上市，分别是Opdivo（通用名Nivolumab）和Keytruda（通用名Pembrolizumab），Opdivo已被批准的适应症包括不可切除的或转移性黑色素瘤以及转移性鳞状非小细胞肺癌，Keytruda被批准的适应症有不可切除的或转移性黑色素瘤以及非小细胞肺癌、头颈部鳞状细胞癌、典型霍奇金淋巴瘤、尿路上皮癌、微卫星高不稳定性癌等。
根据美国FDA药品说明书披露的临床试验结果，Opdivo在一项治疗120名不可切除或转移性黑色素瘤患者的临床试验中客观缓解率（ORR）为32%（38人/120人），包括4名完全缓解（CR）和34名部分缓解（PR）的患者；在另一项治疗117名转移性鳞状非小细胞肺癌患者的临床试验中客观缓解率为15%（17人/117人）。Keytruda在一项834名不可切除或转移性黑色素瘤患者参加的随机对照、开放、多中心临床试验中，两个试验组（分别接受不同剂量的Keytruda治疗）的ORR分别为33%（每三周给药10mg/kg）和34%（每两周给药10mg/kg），其中CR率分别为6%、5%，PR率为分别27%、29%，而对照组（抗CTLA-4单抗Ipilimumab治疗）的ORR为12%；在另一项305名转移性非小细胞肺癌患者参加的随机对照、开放、多中心临床试验中，试验组（Keytruda治疗）的ORR为45%（其中CR4%、PR41%），而对照组（化疗）的ORR为28%（其中CR1%、PR27%）。以上研究可以看出，虽然与化疗和抗CTLA-4治疗相比，Opdivo和Keytruda的抗PD-1治疗在客观缓解率ORR上有了显著提高，然而仍然仅有三分之一左右的患者受益，一多半患者不能获得病情缓解。
抗PD-1抗体治疗恶性肿瘤的有效性主要与其对PD-1/PD-L1结合的阻断作用有关。抗PD-1抗体在PD-1分子上的结合表位及其多态性，以及结合的亲和力（一般用平衡解离常数KD值表示）、特异性等多种因素都会影响抗PD-1抗体对PD-1/PD-L1结合的阻断效果。
噬菌体展示抗体库是一种高通量筛选抗体的技术，目前业内常以重组表达的靶分子胞外区作为靶抗原筛选抗体库。这些靶分子的胞外区仅有一个或数个一般由5-15个氨基酸组成功能表位负责与配体结合，因此只有针对特定功能表位的抗体才具有成药潜力。而用于抗体库筛选的重组表达的胞外区片段除个别发挥生物学功能的功能域外，还有大量的优势抗原表位，因此筛选获得的抗体多数仅有结合活性，但却没有阻断其生物学功能的中和活性，没有成药潜力，需再经过大量的功能学实验和表位鉴定工作才能选出具有治疗作用的功能性抗体。也有采用合成的表位肽筛选抗体库的案例，然而由于合成的表位肽与天然构象的抗原表位可能存在差异，能与表位肽结合的抗体不一定能与活细胞表面的抗原表位结合。总之，现有抗体库筛选技术对有成药潜力的表位特异的功能性抗体命中率低，严重影响了高通量筛选的效率，增加了后期克隆鉴定的成本。
发明内容
本发明提供了一种高效筛选抗体库的方法，能够解决现有抗体库筛选技术对有成药潜力的表位特异的功能性抗体命中率低的问题。本发明还提供了一种采用这种筛选方法获得的抗PD-1单克隆抗体，与Nivolumab、Pembrolizumab拥有不同的结合表位，并且亲和力高、特异性好，可解决部分患者对现有抗PD-1治疗无效的问题。
本发明的技术方案如下：
一种表位特异的功能性抗体的筛选方法，包括以下两个筛选步骤：
步骤一：针对具有生物学功能的抗原表位，合成抗原表位肽并偶联到固相介质上，从噬菌体展示抗体库中进行首轮筛选，获得与表位肽特异性结合的噬菌体并扩增；
步骤二：获取或构建在细胞表面高表达目的抗原的细胞株，用活细胞从首轮筛选获得的噬菌体中进一步筛选与细胞表面抗原结合的噬菌体，用以制备功能性抗体。
为增强筛选的特异性，可步骤一、步骤二序贯循环筛选2-5轮，按本技术方案进行抗体筛选的技术流程图可参考附图1。
步骤一中，具有生物学功能的抗原表位的氨基酸序列可根据蛋白质结晶、片段缺失/点突变对配体/受体的亲和力分析、软件预测等方法获得，所述的抗原表位肽是由10至15个氨基酸组成的环肽，环肽的氨基酸构成符合以下条件：
（1）环肽中有且仅有1个半胱氨酸，环肽通过该半胱氨酸的巯基共价偶联到固相介质上；
（2）环肽中半胱氨酸的两侧各有一段由2个或3个氨基酸构成的连接肽，构成连接肽的氨基酸只能是甘氨酸或脯氨酸；
（3）环肽中含有一段由5-8个氨基酸组成的抗原表位。
优选的，环肽中与半胱氨酸氨基端相连的一段连接肽完全由甘氨酸构成，与半胱氨酸羧基端相连的一段连接肽中含有1到2个脯氨酸。
具体而言，当用于筛选抗人PD-1第75-82位氨基酸所构成的功能表位（氨基酸序列为QTDKLAAF）的单克隆抗体时，步骤一所使用的抗原表位肽是环状并且符合SEQ ID NO: 13所记载的序列；步骤二所使用的细胞株是在细胞表面高表达人PD-1的细胞株，优选的，该细胞株是采用基因重组技术改造的表达外源人PD-1基因的细胞株，而该细胞株天然状态下（即基因改造之前）不表达PD-1，优选的，人PD-1在细胞表面的表达密度为10,000个/细胞到500,000个/细胞。
一种采用上述方法筛选到的抗体，能特异性的结合人PD-1分子从N端开始第75-82位氨基酸（氨基酸序列为QTDKLAAF），该抗体的重链的互补决定区CDR1、CDR2、CDR3分别具有SEQ ID NO:1、SEQ ID NO: 2、SEQ ID NO: 3的氨基酸序列，轻链的互补决定区CDR1、CDR2、CDR3分别具有SEQ ID NO:4、SEQ ID NO: 5、SEQ ID NO: 6的氨基酸序列。
优选的，重链可变区具有SEQ ID NO: 7的氨基酸序列，轻链可变区具有SEQ IDNO: 8的氨基酸序列。
优选的，重链为IgG4亚型，具有SEQ ID NO: 9的氨基酸序列，轻链具有SEQ ID NO:10的氨基酸序列。
优选的，重链由SEQ ID NO: 11的核苷酸序列编码，轻链由SEQ ID NO: 12的核苷酸序列编码。
该抗体由中国仓鼠卵巢细胞（CHO细胞）分泌表达。
竞争性抑制试验（实施例6）和点突变试验（实施例7）表明该抗体与Nivolumab和Pembrolizumab有不同的结合表位，对不能与Pembrolizumab结合的D85G点突变PD-1也能结合。
该抗体有较Nivolumab和Pembrolizumab更低的IC50值（实施例5），能增强人T淋巴细胞对SEB刺激应答能力（实施例8），能在低剂量下更有效的阻断PD-1与PD-L1的结合，并且在较低的剂量下对恶性肿瘤起到治疗效果（实施例9）。
该抗体可单独或与其他药物、辅料组合用于制备恶性肿瘤治疗药物和免疫系统疾病治疗药物。
本发明的有益效果包括，采用本发明的抗体库筛选方法，能够将对表位特异的功能性抗体的命中率提高到30%以上，大大减少克隆鉴定的数量和成本。本发明的抗PD-1单克隆抗体与Nivolumab、Pembrolizumab拥有不同的人PD-1结合表位，对不能与Pembrolizumab结合的点突变PD-1也能结合，并且与PD-1亲和力高，有较低的IC50值，在低剂量下更有效的阻断PD-1与PD-L1的结合，并且在较低的剂量下对恶性肿瘤起到治疗效果，而低剂量抗体的应用对降低药物对机体的刺激、降低药物成本和价格有积极意义。
附图说明
图1 表位特异的功能性抗体筛选技术路线图
图2 实施例5全人源抗PD-1单克隆抗体5L12对PD-1/PD-L1结合的竞争抑制实验结果图
图3 实施例6其他抗PD-1抗体对全人源抗PD-1单克隆抗体5L12与PD-1结合的竞争抑制实验结果图
图4 实施例7 ELISA测试全人源抗PD-1单克隆抗体5L12与D85G突变的PD-1的结合能力实验结果图
图5 实施例8全人源抗PD-1单克隆抗体5L12增强人T淋巴细胞对SEB刺激应答能力的测试实验结果图
图6 实施例9全人源抗PD-1单克隆抗体5L12在动物模型中的抑瘤实验高剂量组结果图
图7 实施例9全人源抗PD-1单克隆抗体5L12在动物模型中的抑瘤实验低剂量组结果图。
具体实施方式
实施例1、合成PD-1表位肽筛选噬菌体展示抗体库
（1）表位肽合成与磁珠偶联
按照SEQ ID NO: 13的序列化学合成人PD-1环状表位肽（委托生工生物工程（上海）股份有限公司合成），该表位肽包括人PD-1第75-82位氨基酸QTDKLAAF，以及供磁珠偶联的半胱氨酸M及其两边的各由3个氨基酸组成的连接肽，分子量1475 Da。将合成好的表位肽以10mg/mL溶解于PBS（磷酸盐缓冲液，pH7.2，含20mM磷酸钠盐，150mM氯化钠）中备用；
将2mL氨基衍生磁珠（Thermofisher Scientific公司，货号21352，每mL磁珠重量为2.5g，含大约12 µmol氨基）用2mL PBS清洗，清洗三遍，磁力分离磁珠；
将Sulfo-SMCC（Thermofisher Scientific公司，货号22122）以4.8mg/mL的浓度溶解于PBS，取200 μL与上述磁珠混合，4℃反应2小时；
分离磁珠并用PBS清洗，将磁珠分为两份，一份重悬于1mL含1%BSA的PBST（含0.1%Tween 20的PBS）中，记为磁珠A；另一份加入5 mL环状表位肽，4℃反应2小时，将磁珠分离用PBST清洗，重悬于1mL含1%BSA的PBST中，记为磁珠B。
（2）筛选噬菌体展示抗体库
收集50名健康志愿者血液各20 mL，混合，按文献《天然人源IgG Fab噬菌体抗体库的构建及多样性分析》（中国免疫学杂志，2010，26 (11) :1007-1010）的方法构建噬菌体展示抗体库。
将步骤（1）准备的磁珠A与1mL用含1%BSA的PBST稀释的的噬菌体展示抗体库（5×10^12 PFU）混合，于室温下旋转混合1小时，磁力去除磁珠A，再加入步骤（1）中准备的磁珠B，于室温下旋转混合1小时，磁力分离磁珠B，用PBST将磁珠B清洗3次，加入0.2 mol/LGlycine-HCl (pH 2.2)溶液温和振动20 min将磁珠B上的噬菌体洗下收集，再用1mol/LTris-HCl（pH 9.1）中和，将获得的噬菌体感染大肠杆菌（E. coli）进行扩增，成为次级库，储存备用。
实施例2、PD-1高表达活细胞二次筛选功能性抗体
（1）构建在细胞表面高表达PD-1的细胞株
克隆全长PD-1编码基因（PDCD1基因，含有N端信号肽编码序列，具体序列见NCBIReference Sequence: NM_005018.2），插入pcDNA3.1表达载体，在大肠杆菌（E. coli）中扩增后转染293T细胞（ATCC编号CRL-3216），筛选高表达细胞株，按照文献“Flow cytometricdetection and quantitation of the epidermal growth factor receptor incomparison to Scatchard analysis in human bladder carcinoma cell lines.Cytometry. 1994 Sep 1;17(1):75-83”和“Antibody- dependent cellularcytotoxicity mediated by cetuximab against lung cancer cell lines. ClinCancer Res. 2007 Mar 1;13(5):1552-61”记载的流式细胞术法测定各细胞株表面的PD-1表达密度，从中选取表达密度最高的编号为2H-3的细胞株，经测定细胞株2H-3表面PD-1的表达密度平均是12000个/细胞，将细胞采用293 SFM II无血清培养基（ThermofisherScientific公司，货号11686029）悬浮培养，将细胞数量扩增至5×10^6以上。
（2）活细胞筛选抗PD-1抗体
将对数期生长的空白293T细胞以1×10^6细胞/mL的密度重悬于1mL DMEM培养液中，加入实施例1（2）中筛选获得的噬菌体（离心后重悬于1mL DMEM培养液中），混匀后37℃孵育1小时，低速离心去除细胞，再重复该过程1次，以去除与293T细胞结合的无效抗体、降低筛选的背景值；然后在离心的上清液中加入实施例2（1）中制备的高表达PD-1的2H-3细胞5×10^5个，混匀后37℃孵育1小时，低速离心收获细胞，用1.5倍体积76 mM柠檬酸溶液（pH2.5）将噬菌体从细胞上洗下，然后用0.5倍体积Tris-HCl（pH7.4）中和，将获得噬菌体感染大肠杆菌（E. coli）进行扩增。
按实施例1（2）和实施例2（2）的筛选方法对上面获得的噬菌体再依次进行两轮筛选，对最终获得的噬菌体进行单克隆化并扩增，挑选60个克隆采用ELISA法进行噬菌体对PD-1的亲和性和结合表位的鉴定（参考“免疫学杂志2012年6月第28卷第6期：489-496”的Phage-ELISA鉴定方法），分别采用重组表达的人PD-1胞外段全长蛋白（第20-170位氨基酸）和化学合成的人PD-1第75-82位氨基酸（序列QTDKLAAF）包被96孔板，并以化学合成的序列为LDSPDR、LAPKAQ、RSQPGQ等短肽作为负对照包被96孔板，然后加入噬菌体和HRP标记的抗噬菌体抗体，加入HRP底物显色反应检测结合情况；采用流式细胞术鉴定噬菌体对活细胞2H-3表面PD-1的亲和性；获得22个ELISA和流式细胞术鉴定与PD-1结合均呈阳性并结合PD-1特定表位（第75-82位氨基酸QTDKLAAF）的克隆，阳性率达36.67%。
实施例3、构建全人源抗PD-1单克隆抗体表达系统
（1）重链和轻链可变区的鉴定
从实施例2最终获得的22个噬菌体阳性克隆中，挑选出5个经ELISA鉴定亲和性较高的克隆进行测序，优选出1个编号为5L12的克隆进行进一步开发。测序结果表明，5L12的重链可变区具有SEQ ID NO: 7的氨基酸序列，轻链可变区具有SEQ ID NO: 8的氨基酸序列，进一步抗体序列分析表明，重链的互补决定区CDR1、CDR2、CDR3分别具有SEQ ID NO:1、SEQ IDNO: 2、SEQ ID NO: 3的氨基酸序列，轻链的互补决定区CDR1、CDR2、CDR3分别具有SEQ IDNO:4、SEQ ID NO: 5、SEQ ID NO: 6的氨基酸序列。
（2）表达载体的构建与转染
以5L12的轻重链可变区基因序列为模板，通过重叠 PCR合成全人源抗体的重链基因（IgG4型）、轻链基因，其中重链的编码序列为SEQ ID NO: 11的核苷酸序列，重链5’端信号肽的编码序列为SEQ ID NO: 14的核苷酸序列，轻链的编码序列为SEQ ID NO: 12的核苷酸序列。将重链、轻链的编码序列插入pcDNA3.1(+)载体（Invitrogen公司），构建全人源抗PD-1单克隆抗体的表达载体pcDNA3.1-anti-PD-1，用Lipofectamine2000Reagent试剂盒进行转染CHO-K1 细胞（ATCC编号CRL-9618），用G418和Zeocin筛选阳性克隆，然后取细胞培养上清液用ELISA方法筛选高表达细胞株，转染、筛选的具体实验过程按照专利CN201010125241（授权公告号CN102167742B）的实施例进行。
（3）小量表达纯化
将筛选得到的高表达细胞株用CHO细胞无血清培养基扩大培养，用 Protein A 亲和柱（GE 公司产品）分离纯化全人源抗体5L12，将纯化的抗体用 PBS 进行透析，最后以紫外吸收法定量。
实施例4、全人源抗PD-1单克隆抗体5L12亲和力的确定
采用表面等离子共振法（SPR）测定实施例3制备的全人源抗PD-1单克隆抗体5L12对人PD-1的亲和力。采用BIACORE3000系统，用经纯化的5L12抗体偶联传感芯片，按照系统的操作说明书进行操作，分别检测5L12与CHO细胞重组表达的PD-1胞外段全长蛋白（NCBI参考序列NM_005018.2第20-170位氨基酸）和化学合成的PD-1第75-82位氨基酸（序列QTDKLAAF）以及序列为LDSPDR、LAPKAQ、RSQPGQ的短肽的亲和力，检测结果见表1，5L12对PD-1胞外段全长及表位肽QTDKLAAF的亲和力达10-11M级。
表1 SPR法测定全人源抗PD-1单克隆抗体5L12与不同肽段亲和性的结果
实施例5、全人源抗PD-1单克隆抗体5L12对PD-1/PD-L1结合的竞争抑制实验
用0.025M pH9.5的碳酸盐缓冲液将待标记蛋白PD-L1（CHO细胞重组表达的人PD-L1胞外段）稀释成1%（质量体积比）浓度，装入透析袋中。用同一缓冲液将FITC配成0.1mg/ml的溶液盛于小烧杯中，使透析袋浸没于FITC溶液中，在4℃避光搅拌24h。取出透析袋中标记液，用Sephadex G-50过柱，去除游离荧光素，收集荧光抗体 FITC-PD-L1备用。
将表面表达人PD-1的基因重组293T细胞以1×106细胞/ml的密度重悬于pH7.2的PBS溶液中；将固定的亚饱和浓度的荧光标记蛋白FITC-PD-L1分别和以4×系列稀释的未标记纯化抗体5L12、Nivolumab（Opdivo）、Pembrolizumab（Keytruda）、人IgG（对照）混合，加入到细胞悬液中，在4 ℃孵育60 min，离心收获细胞，用1% FCS-PBS缓冲液洗2遍，流式细胞仪检测细胞荧光强度并用Cellquest软件分析。竞争抗体的每个浓度设3个复管，计算半数抑制浓度IC50值，最大荧光强度表示在没有竞争抗体时获得的平均荧光强度。
实验结果见图2，表明抗体5L12能完全阻断荧光标记抗体FITC-PD-L1与表面表达PD-1的293T细胞的结合，其IC50值约为30 ng/mL，分别是Nivolumab、Pembrolizumab的1/20、1/2，表明5L12能够更有效的阻断PD-1与其配体的结合，能够发挥效果的最小剂量更低。
实施例6、其他抗PD-1抗体对全人源抗PD-1单克隆抗体5L12与PD-1结合的竞争抑制实验
用实施例5的方法对5L12进行FITC标记，制备FITC-5L12荧光抗体备用。
将表面表达人PD-1的基因重组293T细胞以1×106细胞/ml的密度重悬于pH7.2的PBS溶液中；将固定的亚饱和浓度的荧光标记蛋白FITC-5L12分别和以10×系列稀释的未标记抗体Nivolumab（Opdivo）、Pembrolizumab（Keytruda）、人IgG（对照）混合，加入到细胞悬液中，在4 ℃孵育60 min，离心收获细胞，用1% FCS-PBS缓冲液洗2遍，流式细胞仪检测细胞荧光强度并用Cellquest软件分析。竞争抗体的每个浓度设3个复管，最大荧光强度表示在没有竞争抗体时获得的平均荧光强度。
实验结果见图3，结果表明Pembrolizumab对5L12与人PD-1的结合有竞争抑制作用，而Nivolumab对5L12没有明显的竞争抑制作用，这说明5L12在人PD-1上的结合表位与Nivolumab（主要结合表位为25LDSPDR30，参考文献：An unexpected N-terminal loop inPD-1 dominates binding by nivolumab. Nature Communications, 2017, 8: 14369）不同，但与Pembrolizumab（主要结合表位为Pro83～Gly90，参考文献：High-resolutioncrystal structure of the therapeutic antibody pembrolizumab bound to thehuman PD-1. Scientific Reports. 2016, 6, Article number: 35297）靠近，或者Pembrolizumab与PD-1的结合在空间上阻碍了5L12与PD-1的结合。
实施例7、全人源抗PD-1单克隆抗体5L12与D85G突变的PD-1结合的能力
通过基因重组的方法将人PD-1第85位天冬氨酸突变为甘氨酸（D85G突变），将人PD-1胞外段和人PD-1胞外段（D85G）编码基因分别插入pcDNA3.1载体并转染CHO细胞，进行瞬时表达、纯化。
将纯化的浓度为1 mg/mL的人PD-1胞外段和人PD-1胞外段（D85G）蛋白以100 μL/孔分别包被96孔板，将5L12从1 mg/mL开始进行10×系列稀释，以100 μL/孔加入96孔板，然后加入HRP标记的兔抗人IgG，进行显色反应，用酶标仪读取数据。ELISA结果见图4，结果表明5L12与人PD-1胞外段和人PD-1胞外段（D85G）均能结合，而Pembrolizumab报道不能与D85G突变的PD-1结合（参考文献：Structural basis for blocking PD-1-mediatedimmune suppression by therapeutic antibody pembrolizumab. Cell Research 2017.27:147-150.），说明5L12与Pembrolizumab有不同的结合位点。
实施例8、全人源抗PD-1单克隆抗体5L12增强人T淋巴细胞对SEB刺激应答能力的测试
将来自健康志愿者的外周血以1∶10稀释到细胞培养基中，全血铺板（每孔150 μL）到96-孔板中，用10×系列稀释的全人源抗PD-1单克隆抗体5L12预孵育30-60分钟，然后加入1μg/mL的葡萄球菌肠毒素B（SEB），培养2-4天，收集上清液，用ELISA测量其中IL-2含量，结果见图5，表明5L12能提高全血细胞在SEB刺激下的IL-2产生，并且IL-2水平与5L12的添加剂量具有相关性。
实施例9、全人源抗PD-1单克隆抗体5L12在动物模型中的抑瘤实验
1、人PBMC免疫重建小鼠的构建
使用人淋巴细胞分离液，分离来自健康供者人PBMC（外周血单个核细胞），以8×107/mL重悬于RPMI-1640培养基中备用；选用血清中鼠IgG<5μg/mL的NOD/SCID成年小鼠，尾静脉注射2×107的PBMC细胞，分别在第0、14、28、42、56天从小鼠眼眶后静脉丛取外周血250μL，使用PE标记的鼠抗体人CD3单抗、FITC标记的鼠抗人CD19单抗结合流式细胞仪检测CD3+T细胞、CD19+B细胞含量，利用ELISA法测定小鼠血清中人IgG含量，以确定免疫重建效果。
2、构建小鼠肿瘤模型并利用抗人PD-1抗体5L12治疗
人非小细胞肺癌细胞株HCC827（高表达PD-L1）以1×106细胞/只的剂量皮下植入到人PBMC免疫重建小鼠，植入后待肿瘤生长至250mm2后开始给予抗PD-1治疗。将小鼠分为高剂量组和低剂量组，高剂量组分别给予3 mg/kg的5L12（以1 mg/mL的浓度溶解于PBS中）、Nivolumab、Pembrolizumab，每10天给药1次，给药4次，每隔1周用测径器测量肿瘤大小；低剂量组的给药剂量均调整为0.5 mg/kg，其他试验方法不变；并用PBS作为对照。
实验结果见图6和图7，表明5L12在低剂量下就能对肿瘤生长起到明显的抑制作用，起效剂量明显低于Nivolumab、Pembrolizumab。
序列表
<110> 泰州迈博太科药业有限公司
<120> 一种表位特异的抗体筛选方法及所筛选到的抗体
<130> 2017
<160> 14
<170> SIPOSequenceListing 1.0
<210> 1
<211> 7
<212> PRT
<213> 人工序列()
<400> 1
Gly Tyr Thr Phe Thr Ser Tyr
1 5
<210> 2
<211> 6
<212> PRT
<213> 人工序列()
<400> 2
Tyr Pro Ser Thr Gly Tyr
1 5
<210> 3
<211> 8
<212> PRT
<213> 人工序列()
<400> 3
Gly Val Arg Tyr Tyr Phe Asp Tyr
1 5
<210> 4
<211> 17
<212> PRT
<213> 人工序列()
<400> 4
Lys Ser Ser Gln Ser Leu Leu Asp Ser Ser Thr Arg Lys Asn Tyr Leu
1 5 10 15
Ala
<210> 5
<211> 7
<212> PRT
<213> 人工序列()
<400> 5
Trp Ala Ser Thr Arg Glu Ser
1 5
<210> 6
<211> 7
<212> PRT
<213> 人工序列()
<400> 6
Lys Gln Ser Tyr Asn Leu Trp
1 5
<210> 7
<211> 117
<212> PRT
<213> 人工序列()
<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 Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Met Leu Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Tyr Ile Tyr Pro Ser Thr Gly Tyr Thr Glu Tyr Ser Glu Asn Phe
50 55 60
Lys Asp 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 Arg Gly Val Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 8
<211> 113
<212> PRT
<213> 人工序列()
<400> 8
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser
20 25 30
Ser Thr Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys
35 40 45
Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln
85 90 95
Ser Tyr Asn Leu Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
Arg
<210> 9
<211> 444
<212> PRT
<213> 人工序列()
<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 Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Met Leu Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Tyr Ile Tyr Pro Ser Thr Gly Tyr Thr Glu Tyr Ser Glu Asn Phe
50 55 60
Lys Asp 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 Arg Gly Val Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro
210 215 220
Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
245 250 255
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
260 265 270
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
275 280 285
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
290 295 300
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
305 310 315 320
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
325 330 335
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
340 345 350
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
355 360 365
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
370 375 380
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
385 390 395 400
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
405 410 415
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
420 425 430
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440
<210> 10
<211> 219
<212> PRT
<213> 人工序列()
<400> 10
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser
20 25 30
Ser Thr Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys
35 40 45
Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln
85 90 95
Ser Tyr Asn Leu Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
130 135 140
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
145 150 155 160
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
165 170 175
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
180 185 190
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
195 200 205
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 11
<211> 1332
<212> DNA
<213> 人工序列()
<400> 11
gaggtgcagc tggtggagtc cggcggcggc ctggtgcagc ccggcggctc cctgcggctg 60
tcctgcgccg cctccggcta caccttcacc tcctactgga tgctgtgggt gcggcaggcc 120
cccggcaagg gcctggagtg ggtgtcctac atctacccct ccaccggcta caccgagtac 180
tccgagaact tcaaggaccg gttcaccatc tcccgggaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgcgc ccggggcgtg 300
cggtactact tcgactactg gggccagggc accctggtga ccgtgtcctc cgcctccacc 360
aagggcccct ccgtgttccc cctggccccc tgctcccggt ccacctccga gtccaccgcc 420
gccctgggct gcctggtgaa ggactacttc cccgagcccg tgaccgtgtc ctggaactcc 480
ggcgccctga cctccggcgt gcacaccttc cccgccgtgc tgcagtcctc cggcctgtac 540
tccctgtcct ccgtggtgac cgtgccctcc tcctccctgg gcaccaagac ctacacctgc 600
aacgtggacc acaagccctc caacaccaag gtggacaagc gggtggagtc caagtacggc 660
cccccctgcc ccccctgccc cgcccccgag ttcctgggcg gcccctccgt gttcctgttc 720
ccccccaagc ccaaggacac cctgatgatc tcccggaccc ccgaggtgac ctgcgtggtg 780
gtggacgtgt cccaggagga ccccgaggtg cagttcaact ggtacgtgga cggcgtggag 840
gtgcacaacg ccaagaccaa gccccgggag gagcagttca actccaccta ccgggtggtg 900
tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960
tccaacaagg gcctgccctc ctccatcgag aagaccatct ccaaggccaa gggccagccc 1020
cgggagcccc aggtgtacac cctgcccccc tcccaggagg agatgaccaa gaaccaggtg 1080
tccctgacct gcctggtgaa gggcttctac ccctccgaca tcgccgtgga gtgggagtcc 1140
aacggccagc ccgagaacaa ctacaagacc accccccccg tgctggactc cgacggctcc 1200
ttcttcctgt actcccggct gaccgtggac aagtcccggt ggcaggaggg caacgtgttc 1260
tcctgctccg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgtccctg 1320
tccctgggca ag 1332
<210> 12
<211> 657
<212> DNA
<213> 人工序列()
<400> 12
gacatccaga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgggtgacc 60
atcacctgca agtcctccca gtccctgctg gactcctcca cccggaagaa ctacctggcc 120
tggtaccagc agaagcccgg caaggccccc aagctgctga tctactgggc ctccacccgg 180
gagtccggcg tgccctcccg gttctccggc tccggctccg gcaccgactt caccctgacc 240
atctcctccc tgcagcccga ggacttcgcc acctactact gcaagcagtc ctacaacctg 300
tggaccttcg gccagggcac caaggtggag atcaagcgga ccgtggccgc cccctccgtg 360
ttcatcttcc ccccctccga cgagcagctg aagtccggca ccgcctccgt ggtgtgcctg 420
ctgaacaact tctacccccg ggaggccaag gtgcagtgga aggtggacaa cgccctgcag 480
tccggcaact cccaggagtc cgtgaccgag caggactcca aggactccac ctactccctg 540
tcctccaccc tgaccctgtc caaggccgac tacgagaagc acaaggtgta cgcctgcgag 600
gtgacccacc agggcctgtc ctcccccgtg accaagtcct tcaaccgggg cgagtgc 657
<210> 13
<211> 15
<212> PRT
<213> 人工序列()
<400> 13
Cys Pro Pro Ala Gln Thr Asp Lys Leu Ala Ala Phe Ala Ala Ala
1 5 10 15
<210> 14
<211> 66
<212> DNA
<213> 人工序列()
<400> 14
atggattttc aggtgcagat tttcagcttc ctgctaatca gtgcctcagt cataatatcc 60
agagga 66
Claims (10)
1.一种表位特异的功能性抗体的筛选方法，其特征是，包括以下两个筛选步骤：
步骤一：合成抗原表位肽并偶联到固相介质上，从噬菌体展示抗体库中进行首轮筛选，获得与表位肽特异性结合的噬菌体并扩增；
步骤二：获取或构建在细胞表面高表达目的抗原的细胞株，用活细胞从首轮筛选获得的噬菌体中进一步筛选与细胞表面抗原结合的噬菌体。
2.根据权利要求1所述的方法，其特征是，步骤一所述的抗原表位肽是由10至15个氨基酸组成的环肽，环肽的氨基酸构成符合以下条件：
环肽中有且仅有1个半胱氨酸，环肽通过该半胱氨酸的巯基共价偶联到固相介质上；
环肽中半胱氨酸的两侧各有一段由2个或3个氨基酸构成的连接肽，构成连接肽的氨基酸只能是甘氨酸或脯氨酸；
环肽中含有一段由5-8个氨基酸组成的抗原表位。
3.根据权利要求2所述的方法，其特征是，环肽中与半胱氨酸氨基端相连的一段连接肽完全由甘氨酸构成，与半胱氨酸羧基端相连的一段连接肽中含有1到2个脯氨酸。
4.一种表位特异的抗人PD-1功能性抗体的筛选方法，其特征是，采用权利要求1所述的方法，并且步骤一所使用的抗原表位肽是具有SEQ ID NO: 13所记载的氨基酸序列的环肽，步骤二所使用的细胞株是细胞表面高表达PD-1的细胞株，PD-1在细胞表面的表达数量为10,000个/细胞到500,000个/细胞。
5.一种采用权利要求1所述的方法筛选到的抗体，能特异性的结合人PD-1分子的胞外段，其特征是，重链的互补决定区CDR1、CDR2、CDR3分别具有SEQ ID NO:1、SEQ ID NO: 2、SEQ ID NO: 3的氨基酸序列，轻链的互补决定区CDR1、CDR2、CDR3分别具有SEQ ID NO:4、SEQ ID NO: 5、SEQ ID NO: 6的氨基酸序列。
6.根据权利要求5所述的抗体，其特征是，重链可变区具有SEQ ID NO: 7的氨基酸序列，轻链可变区具有SEQ ID NO: 8的氨基酸序列。
7.根据权利要求5所述的抗体，其特征是，重链具有SEQ ID NO: 9的氨基酸序列，轻链具有SEQ ID NO: 10的氨基酸序列。
8.根据权利要求5所述的抗体，其特征是，重链由SEQ ID NO: 11的核苷酸序列编码，轻链由SEQ ID NO: 12的核苷酸序列编码。
9.一种组合物，含有权利要求6所述的单克隆抗体，和药学上可接受的载体。
10.权利要求6-9所述的抗体在制备恶性肿瘤治疗药物和免疫系统疾病治疗药物中的用途。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711375292.7A CN109929036B (zh) | 2017-12-19 | 2017-12-19 | 一种表位特异的抗体筛选方法及所筛选到的抗体 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711375292.7A CN109929036B (zh) | 2017-12-19 | 2017-12-19 | 一种表位特异的抗体筛选方法及所筛选到的抗体 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109929036A true CN109929036A (zh) | 2019-06-25 |
| CN109929036B CN109929036B (zh) | 2024-12-03 |
Family
ID=66983728
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711375292.7A Active CN109929036B (zh) | 2017-12-19 | 2017-12-19 | 一种表位特异的抗体筛选方法及所筛选到的抗体 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109929036B (zh) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110656092A (zh) * | 2019-11-06 | 2020-01-07 | 新乡学院 | 一种细胞膜表面展示含双信号肽pd-1蛋白的细胞模型及其应用 |
| WO2021043337A1 (zh) * | 2019-09-06 | 2021-03-11 | 中国药科大学 | 一种多肽及其应用 |
| WO2021197212A1 (zh) * | 2020-03-30 | 2021-10-07 | 华东理工大学 | 噬菌体药物蛋白展示系统及其应用 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1649900A (zh) * | 2000-12-29 | 2005-08-03 | 生物技术通用公司 | 分离的包括含硫酸根部分的表位的分子,抗这样的表位的抗体,和它们的应用 |
| US20060275288A1 (en) * | 2005-01-20 | 2006-12-07 | Grihalde Nelson D | GLP-1 receptor agonist and allosteric modulator monoclonal antibodies and uses thereof |
| CN104292308A (zh) * | 2014-08-22 | 2015-01-21 | 广州安辰医药科技有限公司 | 新型肿瘤抗原环肽 |
| CN104447993A (zh) * | 2013-09-13 | 2015-03-25 | 上海海思太科药业有限公司 | 一种高活性全人源抗her2单克隆抗体、其制备方法及用途 |
| US20190231872A1 (en) * | 2016-08-17 | 2019-08-01 | Industry Academic Cooperation Foundation, Hallym University | A vaccine composition comprising cyclic peptides, antibodies to the cyclic peptides or an anticancer composition comprising the same |
-
2017
- 2017-12-19 CN CN201711375292.7A patent/CN109929036B/zh active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1649900A (zh) * | 2000-12-29 | 2005-08-03 | 生物技术通用公司 | 分离的包括含硫酸根部分的表位的分子,抗这样的表位的抗体,和它们的应用 |
| US20060275288A1 (en) * | 2005-01-20 | 2006-12-07 | Grihalde Nelson D | GLP-1 receptor agonist and allosteric modulator monoclonal antibodies and uses thereof |
| CN104447993A (zh) * | 2013-09-13 | 2015-03-25 | 上海海思太科药业有限公司 | 一种高活性全人源抗her2单克隆抗体、其制备方法及用途 |
| CN104292308A (zh) * | 2014-08-22 | 2015-01-21 | 广州安辰医药科技有限公司 | 新型肿瘤抗原环肽 |
| US20190231872A1 (en) * | 2016-08-17 | 2019-08-01 | Industry Academic Cooperation Foundation, Hallym University | A vaccine composition comprising cyclic peptides, antibodies to the cyclic peptides or an anticancer composition comprising the same |
Non-Patent Citations (3)
| Title |
|---|
| SANG JICK KIM等: "Selection of an affinity-matured antibody against a defined epitope by phage display of an immune antibody library", 《JOURNAL OF IMMUNOLOGICAL METHODS》, vol. 329, no. 1, 9 November 2007 (2007-11-09) * |
| 宋宏彬等: "人源抗HIV-1 V3环噬菌体抗体筛选与鉴定", 《中国公共卫生》, vol. 24, no. 7, 1 September 2008 (2008-09-01) * |
| 罗龙龙: "基于表位定向哺乳动物细胞抗体库筛选新型抗PD-1抗体", 《中国知网》, 31 December 2016 (2016-12-31) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021043337A1 (zh) * | 2019-09-06 | 2021-03-11 | 中国药科大学 | 一种多肽及其应用 |
| CN110656092A (zh) * | 2019-11-06 | 2020-01-07 | 新乡学院 | 一种细胞膜表面展示含双信号肽pd-1蛋白的细胞模型及其应用 |
| WO2021197212A1 (zh) * | 2020-03-30 | 2021-10-07 | 华东理工大学 | 噬菌体药物蛋白展示系统及其应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109929036B (zh) | 2024-12-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110229232B (zh) | 双特异性抗体及其用途 | |
| WO2020009725A1 (en) | Human anti-cd47 antibodies and uses thereof | |
| CN119161491A (zh) | 双特异性重组蛋白及其应用 | |
| EA030147B1 (ru) | Биспецифические антигенсвязывающие молекулы, активирующие т-клетки | |
| CN112409483A (zh) | 抗pd-l1纳米抗体 | |
| CN113924316B (zh) | 抗cd25抗体及其应用 | |
| KR20230169944A (ko) | Mage-a4 펩티드-mhc 항원 결합 단백질 | |
| WO2022190007A1 (en) | Rabbit-derived antigen binding protein nucleic acid libraries | |
| WO2021219127A1 (zh) | 一种靶向her2和pd-1的双特异性抗体及其应用 | |
| WO2019242619A1 (zh) | 全人源的抗lag-3抗体及其应用 | |
| CN106164094A (zh) | 双特异性抗原结合多肽 | |
| KR20220087466A (ko) | 항-케모킨(anti-chemokin) 유사 수용체 1 사람화된 항체 및 이의 치료학적 적용 | |
| CN109929036B (zh) | 一种表位特异的抗体筛选方法及所筛选到的抗体 | |
| JP2025534237A (ja) | Ｍａｇｅ－ａ４ペプチドデュアルｔ細胞エンゲージャー | |
| WO2023217216A1 (zh) | 一种三特异性抗体的设计、制备及用途 | |
| CN113603779B (zh) | 一种结合人淋巴细胞活化基因3(lag-3)的抗体及其用途 | |
| WO2019192493A1 (zh) | 抗人lag-3单克隆抗体及其应用 | |
| CN111378040B (zh) | 检测多种恶性肿瘤细胞的抗体及其应用 | |
| CN111333732B (zh) | 一种靶向人bcma且激活nk细胞的双特异性抗体的制备及其应用 | |
| CN114163528A (zh) | Cd47结合分子及其应用 | |
| TW202227504A (zh) | 抗ｅｒｂｂ３抗體或其抗原結合片段及其醫藥用途 | |
| WO2021244371A1 (zh) | 一种抗pd-l1/vegf融合蛋白 | |
| WO2021244587A1 (zh) | 一种抗PD-L1/TGF-β融合蛋白 | |
| WO2021083248A1 (zh) | 抗tspan8单克隆抗体及其用途 | |
| CN114773485B (zh) | 抗人PD-L1抗体和TGFβRII的双功能融合蛋白分子 |
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 | ||
| GR01 | Patent grant |