CN107286237B - 一种抗丙型肝炎病毒抗体的获取以及应用 - Google Patents
一种抗丙型肝炎病毒抗体的获取以及应用 Download PDFInfo
- Publication number
- CN107286237B CN107286237B CN201710668058.7A CN201710668058A CN107286237B CN 107286237 B CN107286237 B CN 107286237B CN 201710668058 A CN201710668058 A CN 201710668058A CN 107286237 B CN107286237 B CN 107286237B
- Authority
- CN
- China
- Prior art keywords
- antibody
- hcv
- seq
- antigen
- binding fragment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000700605 Viruses Species 0.000 title claims abstract description 16
- 208000005176 Hepatitis C Diseases 0.000 title claims abstract description 7
- 230000027455 binding Effects 0.000 claims abstract description 31
- 239000012634 fragment Substances 0.000 claims abstract description 30
- 239000000427 antigen Substances 0.000 claims abstract description 28
- 102000036639 antigens Human genes 0.000 claims abstract description 28
- 108091007433 antigens Proteins 0.000 claims abstract description 28
- 230000003472 neutralizing effect Effects 0.000 claims abstract description 27
- 208000015181 infectious disease Diseases 0.000 claims abstract description 22
- 201000010099 disease Diseases 0.000 claims abstract description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 5
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims description 19
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 17
- 239000013604 expression vector Substances 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 7
- 108020004707 nucleic acids Proteins 0.000 claims description 7
- 102000039446 nucleic acids Human genes 0.000 claims description 7
- 150000007523 nucleic acids Chemical class 0.000 claims description 7
- 230000001154 acute effect Effects 0.000 claims description 6
- 239000008194 pharmaceutical composition Substances 0.000 claims description 6
- 208000006454 hepatitis Diseases 0.000 claims description 5
- 231100000283 hepatitis Toxicity 0.000 claims description 5
- 230000002265 prevention Effects 0.000 claims description 5
- 208000019423 liver disease Diseases 0.000 claims description 4
- 208000006154 Chronic hepatitis C Diseases 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 208000010710 hepatitis C virus infection Diseases 0.000 claims description 3
- 206010065051 Acute hepatitis C Diseases 0.000 claims description 2
- 206010016654 Fibrosis Diseases 0.000 claims description 2
- 101000998953 Homo sapiens Immunoglobulin heavy variable 1-2 Proteins 0.000 claims description 2
- 102100036887 Immunoglobulin heavy variable 1-2 Human genes 0.000 claims description 2
- 208000037621 acute hepatitis C virus infection Diseases 0.000 claims description 2
- 230000007882 cirrhosis Effects 0.000 claims description 2
- 208000019425 cirrhosis of liver Diseases 0.000 claims description 2
- 201000007270 liver cancer Diseases 0.000 claims description 2
- 208000014018 liver neoplasm Diseases 0.000 claims description 2
- 101100297347 Caenorhabditis elegans pgl-3 gene Proteins 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 201000001441 melanoma Diseases 0.000 claims 1
- 230000009465 prokaryotic expression Effects 0.000 claims 1
- 238000001228 spectrum Methods 0.000 abstract 1
- 241000711549 Hepacivirus C Species 0.000 description 52
- 210000004027 cell Anatomy 0.000 description 11
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 6
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 210000003719 b-lymphocyte Anatomy 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000006386 neutralization reaction Methods 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 230000003612 virological effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 241000283707 Capra Species 0.000 description 3
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 3
- 108090000288 Glycoproteins Proteins 0.000 description 3
- 102000003886 Glycoproteins Human genes 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- -1 microsomes Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- DXQIQUIQYAGRCC-CIUDSAMLSA-N Arg-Asp-Gln Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)CN=C(N)N DXQIQUIQYAGRCC-CIUDSAMLSA-N 0.000 description 2
- UMHUHHJMEXNSIV-CIUDSAMLSA-N Asp-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UMHUHHJMEXNSIV-CIUDSAMLSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 102100027723 Endogenous retrovirus group K member 6 Rec protein Human genes 0.000 description 2
- 101710121417 Envelope glycoprotein Proteins 0.000 description 2
- YYPFZVIXAVDHIK-IUCAKERBSA-N Gly-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)CN YYPFZVIXAVDHIK-IUCAKERBSA-N 0.000 description 2
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 2
- RGPWUJOMKFYFSR-QWRGUYRKSA-N His-Gly-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O RGPWUJOMKFYFSR-QWRGUYRKSA-N 0.000 description 2
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 108010052285 Membrane Proteins Proteins 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- JEGFCFLCRSJCMA-IHRRRGAJSA-N Phe-Arg-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N JEGFCFLCRSJCMA-IHRRRGAJSA-N 0.000 description 2
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- BSNZTJXVDOINSR-JXUBOQSCSA-N Thr-Ala-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BSNZTJXVDOINSR-JXUBOQSCSA-N 0.000 description 2
- YOOAQCZYZHGUAZ-KATARQTJSA-N Thr-Leu-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YOOAQCZYZHGUAZ-KATARQTJSA-N 0.000 description 2
- RMRFSFXLFWWAJZ-HJOGWXRNSA-N Tyr-Tyr-Tyr 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(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 RMRFSFXLFWWAJZ-HJOGWXRNSA-N 0.000 description 2
- 238000007792 addition Methods 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 230000009260 cross reactivity Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 108010074027 glycyl-seryl-phenylalanine Proteins 0.000 description 2
- 108010089804 glycyl-threonine Proteins 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 238000009169 immunotherapy Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 108010083708 leucyl-aspartyl-valine Proteins 0.000 description 2
- 108010012058 leucyltyrosine Proteins 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 210000001806 memory b lymphocyte Anatomy 0.000 description 2
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000003259 recombinant expression Methods 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- SHYYAQLDNVHPFT-DLOVCJGASA-N Ala-Asn-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 SHYYAQLDNVHPFT-DLOVCJGASA-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
- VNYMOTCMNHJGTG-JBDRJPRFSA-N Ala-Ile-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O VNYMOTCMNHJGTG-JBDRJPRFSA-N 0.000 description 1
- 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 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- ZDILXFDENZVOTL-BPNCWPANSA-N Ala-Val-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZDILXFDENZVOTL-BPNCWPANSA-N 0.000 description 1
- OTOXOKCIIQLMFH-KZVJFYERSA-N Arg-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N OTOXOKCIIQLMFH-KZVJFYERSA-N 0.000 description 1
- OTZMRMHZCMZOJZ-SRVKXCTJSA-N Arg-Leu-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O OTZMRMHZCMZOJZ-SRVKXCTJSA-N 0.000 description 1
- ADPACBMPYWJJCE-FXQIFTODSA-N Arg-Ser-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O ADPACBMPYWJJCE-FXQIFTODSA-N 0.000 description 1
- MRQQMVZUHXUPEV-IHRRRGAJSA-N Asp-Arg-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O MRQQMVZUHXUPEV-IHRRRGAJSA-N 0.000 description 1
- USNJAPJZSGTTPX-XVSYOHENSA-N Asp-Phe-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O USNJAPJZSGTTPX-XVSYOHENSA-N 0.000 description 1
- ZQFRDAZBTSFGGW-SRVKXCTJSA-N Asp-Ser-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZQFRDAZBTSFGGW-SRVKXCTJSA-N 0.000 description 1
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 1
- GYNUXDMCDILYIQ-QRTARXTBSA-N Asp-Val-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(=O)O)N GYNUXDMCDILYIQ-QRTARXTBSA-N 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 102100027207 CD27 antigen Human genes 0.000 description 1
- SZQCDCKIGWQAQN-FXQIFTODSA-N Cys-Arg-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O SZQCDCKIGWQAQN-FXQIFTODSA-N 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000005593 Endopeptidases Human genes 0.000 description 1
- 108010059378 Endopeptidases Proteins 0.000 description 1
- 241000792859 Enema Species 0.000 description 1
- 101800001467 Envelope glycoprotein E2 Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000710831 Flavivirus Species 0.000 description 1
- 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 1
- LJEPDHWNQXPXMM-NHCYSSNCSA-N Gln-Arg-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O LJEPDHWNQXPXMM-NHCYSSNCSA-N 0.000 description 1
- BTSPOOHJBYJRKO-CIUDSAMLSA-N Gln-Asp-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O BTSPOOHJBYJRKO-CIUDSAMLSA-N 0.000 description 1
- LOJYQMFIIJVETK-WDSKDSINSA-N Gln-Gln Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(O)=O LOJYQMFIIJVETK-WDSKDSINSA-N 0.000 description 1
- XZLLTYBONVKGLO-SDDRHHMPSA-N Gln-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(=O)N)N)C(=O)O XZLLTYBONVKGLO-SDDRHHMPSA-N 0.000 description 1
- OTQSTOXRUBVWAP-NRPADANISA-N Gln-Ser-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O OTQSTOXRUBVWAP-NRPADANISA-N 0.000 description 1
- SGVGIVDZLSHSEN-RYUDHWBXSA-N Gln-Tyr-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O SGVGIVDZLSHSEN-RYUDHWBXSA-N 0.000 description 1
- XTZDZAXYPDISRR-MNXVOIDGSA-N Glu-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N XTZDZAXYPDISRR-MNXVOIDGSA-N 0.000 description 1
- INGJLBQKTRJLFO-UKJIMTQDSA-N Glu-Ile-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCC(O)=O INGJLBQKTRJLFO-UKJIMTQDSA-N 0.000 description 1
- YQPFCZVKMUVZIN-AUTRQRHGSA-N Glu-Val-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O YQPFCZVKMUVZIN-AUTRQRHGSA-N 0.000 description 1
- ZYRXTRTUCAVNBQ-GVXVVHGQSA-N Glu-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZYRXTRTUCAVNBQ-GVXVVHGQSA-N 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 102000005720 Glutathione transferase Human genes 0.000 description 1
- 108010070675 Glutathione transferase Proteins 0.000 description 1
- LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 description 1
- OCQUNKSFDYDXBG-QXEWZRGKSA-N Gly-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N OCQUNKSFDYDXBG-QXEWZRGKSA-N 0.000 description 1
- CQZDZKRHFWJXDF-WDSKDSINSA-N Gly-Gln-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CN CQZDZKRHFWJXDF-WDSKDSINSA-N 0.000 description 1
- BHPQOIPBLYJNAW-NGZCFLSTSA-N Gly-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN BHPQOIPBLYJNAW-NGZCFLSTSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- JQFILXICXLDTRR-FBCQKBJTSA-N Gly-Thr-Gly Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)NCC(O)=O JQFILXICXLDTRR-FBCQKBJTSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 description 1
- 101001008255 Homo sapiens Immunoglobulin kappa variable 1D-8 Proteins 0.000 description 1
- 101001047628 Homo sapiens Immunoglobulin kappa variable 2-29 Proteins 0.000 description 1
- 101001008321 Homo sapiens Immunoglobulin kappa variable 2D-26 Proteins 0.000 description 1
- 101001047619 Homo sapiens Immunoglobulin kappa variable 3-20 Proteins 0.000 description 1
- 101001008263 Homo sapiens Immunoglobulin kappa variable 3D-15 Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- IITVUURPOYGCTD-NAKRPEOUSA-N Ile-Pro-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O IITVUURPOYGCTD-NAKRPEOUSA-N 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
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 102100022964 Immunoglobulin kappa variable 3-20 Human genes 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- FEHQLKKBVJHSEC-SZMVWBNQSA-N Leu-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FEHQLKKBVJHSEC-SZMVWBNQSA-N 0.000 description 1
- SBANPBVRHYIMRR-GARJFASQSA-N Leu-Ser-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N SBANPBVRHYIMRR-GARJFASQSA-N 0.000 description 1
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 1
- 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 1
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 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
- 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 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
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 1
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 1
- SJDQOYTYNGZZJX-SRVKXCTJSA-N Met-Glu-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SJDQOYTYNGZZJX-SRVKXCTJSA-N 0.000 description 1
- 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 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 208000037581 Persistent Infection Diseases 0.000 description 1
- 239000004697 Polyetherimide Substances 0.000 description 1
- VCYJKOLZYPYGJV-AVGNSLFASA-N Pro-Arg-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O VCYJKOLZYPYGJV-AVGNSLFASA-N 0.000 description 1
- MGDFPGCFVJFITQ-CIUDSAMLSA-N Pro-Glu-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MGDFPGCFVJFITQ-CIUDSAMLSA-N 0.000 description 1
- 206010037742 Rabies Diseases 0.000 description 1
- LVVBAKCGXXUHFO-ZLUOBGJFSA-N Ser-Ala-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O LVVBAKCGXXUHFO-ZLUOBGJFSA-N 0.000 description 1
- OOKCGAYXSNJBGQ-ZLUOBGJFSA-N Ser-Asn-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O OOKCGAYXSNJBGQ-ZLUOBGJFSA-N 0.000 description 1
- 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 1
- UQFYNFTYDHUIMI-WHFBIAKZSA-N Ser-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CO UQFYNFTYDHUIMI-WHFBIAKZSA-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
- MHVXPTAMDHLTHB-IHPCNDPISA-N Ser-Phe-Trp Chemical compound C([C@H](NC(=O)[C@H](CO)N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 MHVXPTAMDHLTHB-IHPCNDPISA-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
- 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
- LGIMRDKGABDMBN-DCAQKATOSA-N Ser-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N LGIMRDKGABDMBN-DCAQKATOSA-N 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 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
- 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
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 1
- YXSSXUIBUJGHJY-SFJXLCSZSA-N Trp-Thr-Phe Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)[C@H](O)C)C(O)=O)C1=CC=CC=C1 YXSSXUIBUJGHJY-SFJXLCSZSA-N 0.000 description 1
- PKZIWSHDJYIPRH-JBACZVJFSA-N Trp-Tyr-Gln Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O PKZIWSHDJYIPRH-JBACZVJFSA-N 0.000 description 1
- MBLJBGZWLHTJBH-SZMVWBNQSA-N Trp-Val-Arg Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)=CNC2=C1 MBLJBGZWLHTJBH-SZMVWBNQSA-N 0.000 description 1
- QOIKZODVIPOPDD-AVGNSLFASA-N Tyr-Cys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(O)=O QOIKZODVIPOPDD-AVGNSLFASA-N 0.000 description 1
- AKLNEFNQWLHIGY-QWRGUYRKSA-N Tyr-Gly-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N)O AKLNEFNQWLHIGY-QWRGUYRKSA-N 0.000 description 1
- USYGMBIIUDLYHJ-GVARAGBVSA-N Tyr-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 USYGMBIIUDLYHJ-GVARAGBVSA-N 0.000 description 1
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 1
- JQTYTBPCSOAZHI-FXQIFTODSA-N Val-Ser-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N JQTYTBPCSOAZHI-FXQIFTODSA-N 0.000 description 1
- WUFHZIRMAZZWRS-OSUNSFLBSA-N Val-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C(C)C)N WUFHZIRMAZZWRS-OSUNSFLBSA-N 0.000 description 1
- 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 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000033289 adaptive immune response Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 239000000412 dendrimer Substances 0.000 description 1
- 229920000736 dendritic polymer Polymers 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000007920 enema Substances 0.000 description 1
- 229940095399 enema Drugs 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 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
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001553 hepatotropic effect Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Chemical group 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 210000001589 microsome Anatomy 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000007857 nested PCR Methods 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920001601 polyetherimide Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000010741 sumoylation Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
- C07K16/1081—Togaviridae, e.g. flavivirus, rubella virus, hog cholera virus
- C07K16/109—Hepatitis C virus; Hepatitis G virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
-
- 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
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/18—Togaviridae; Flaviviridae
- G01N2333/183—Flaviviridae, e.g. pestivirus, mucosal disease virus, bovine viral diarrhoea virus, classical swine fever virus (hog cholera virus) or border disease virus
- G01N2333/186—Hepatitis C; Hepatitis NANB
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/10—Detection of antigens from microorganism in sample from host
Abstract
本发明涉及一种抗丙型肝炎病毒抗体的获取以及应用，具体涉及作为治疗和预防HCV感染药物的全人源单克隆抗体TRN1007或其功能性片段。TRN1007抗体作为光谱中和抗体，与不同抗原结合能力及不同病毒的中和能力都很强，展示了对所述包括基因型1a、1b、2、3、4、5、6和7的病毒均具有较强的结合活性，特别是对HCV真病毒株S52和HK6a的中和效率，IC50值仅为4.6nM和6.6nM。TRN1007抗体优异的结合、中和活性显示其在检测、治疗和预防HCV感染及其相关疾病的良好应用前景。
Description
技术领域
本发明属于生物医药领域，具体涉及一种全人源抗HCV的单克隆中和抗体及该中和抗体的制备方法和用途。
背景技术
丙型肝炎病毒((Hepatitis C virus，HCV)目前仍是威胁人类健康的一大疾病，影响世界2.0-3.0％的人口。HCV是一种具有外周衣壳(pericapsid)和单链RNA的属于黄病毒家族的病毒。HCV目前根据其基因序列的差异分为7种基因型，每种均以数字标记。每种基因型又进一步分为若干亚型，每种亚型均以字母标记。不同HCV基因型的流行和分布在世界各地不相同。
丙型肝炎病毒是嗜肝病毒，感染肝脏细胞，引起肝脏病变，最近也有一些研究表明HCV感染可以引起肝外一些病理改变。由于HCV感染急性期患者未表现出明显的症状，大部分感染者在被确诊时己经发展为慢性化阶段。HCV感染后约一周感染者体内就能检测到HCVRNA，而获得性免疫应答反应在数月之后才能检测到。对急性期HCV患者的研究表明，广泛且特异性的T细胞免疫应答在清除病毒的过程中具有重要作用。
迄今为止，尚缺乏丙型肝炎病毒有效的疫苗和免疫疗法。尽管被赋予对不同病毒基因型的交叉反应性，HCV抗原结构的高变性至今仍阻碍可中和病毒的抗体的开发。因而迫切需要具有能识别不同基因型病毒特征的并从而在治疗和预防HCV感染中发挥有效作用的抗HCV抗体。
越来越多的证据表明早期且特异性的抗体反应能够影响急性期HCV感染的结果，中和抗体在慢性感染的控制中具有很好的作用。如Burioni等于1998年发表在《Hepatology》第28卷第3期的文章就记载了编码五种特异于HCV E2糖蛋白(HCV E2)的重组人抗体片段(Fab)的序列的克隆和鉴定，所述的抗体片段能够结合不同病毒基因型的糖蛋白(交叉反应性)。随后又发现了能够在体外有效地中和不同HCV基因型的感染的e20抗体片段，但该研究直到现在还没有成功上市。
而本发明提供的全人源抗HCV的单克隆中和抗体，且中和能力很强，由其针对病毒株S52(3a)和HK6a，IC50值仅为4.6nM和6.6nM。TRN1007抗体优异的中和活性显示其在疫苗中的良好应用前景。
发明内容
本发明的目的是提供一种能够结合来自多种不同基因型的HCV病毒的全人源单克隆中和抗体或其抗原结合片段，其作为治疗或预防HCV感染的药剂，其特征在于所述抗体结合HCV表面包膜糖蛋白E2。
进一步，所述重链可变区的互补性决定区域CDR1、CDR2、CDR3和/或轻链可变区的互补性决定区域CDR1、CDR2、CDR3；重链CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4所示；轻链CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8所示。其中所述重链可变区的互补性决定区域和/或轻链可变区的互补性决定区域经过取代、缺失或添加一个或几个氨基酸且形成的抗体或其片段具有相同或相似活性，或者分别与SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4所示的序列至少具有80％同源性；轻链CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:6、SEQ ID NO:7、SEQ IDNO:8所示，或者分别与SEQ ID NO:6、SEQ ID NO:7、SEQID NO:8所示的序列至少具有80％同源性。
进一步，所述抗体或其抗原结合片段包含至少一个包含氨基酸序列SEQ ID NO：1或与SEQ ID NO：1具有至少80％同一性的序列的重链可变区，以及至少一个包含氨基酸序列SEQ ID NO：2或与SEQ ID NO：5具有至少80％同一性的序列的轻链可变区。
进一步优选的序列同一性百分比是至少81％、83％、85％、87％、89％、90％、91％、93％、94％、95％、96％、97％、98％或99％，其中所述表述“至少”是指对于所列的每个百分比。
包括优选抗体氨基酸序列的保守序列变体的抗体也包括在本发明的范围之内。保守氨基酸序列变体包括不显著改变本发明的全人源抗HCV的单克隆中和抗体结合性质和中和性质的氨基酸序列的修饰，如源于本领域熟知的相似氨基酸替代的变体，氨基酸的缺失、增加导致的变体。
本发明的抗体还包括人源与非人源抗体，以及具有与TRN1007抗体相同功能或改造及优化的一切抗体。
进一步，所述抗体的抗原结合片段包括Fab、Fab′、F(ab′)2、Fv、dAb和互补决定区(CDR)片段、单链抗体(例如，scFv)、嵌合抗体、双抗体和这样的多肽，或同时包含重链与轻链可变区或CDRs的单链抗体，或包含来源于免疫球蛋白重链可变区与轻链可变区的CDR片段的一个或更多个拷贝的骨架，其包含足以赋予多肽特异性抗原结合能力的抗体的至少一部分。
术语“抗体”意指任意种类的全长免疫球蛋白及其包含轻链可变区与重链可变区的任意片段，例如Fab、F(ab’)2、CDR(互补决定区)，或同时包含重链与轻链可变区或CDRs的单链抗体，或包含来源于免疫球蛋白重链可变区与轻链可变区的CDR片段的一个或更多个拷贝的骨架。
术语“Fv”意指由抗体的单臂的VL和VH结构域组成的抗体片段；术语“dAb”意指由VH结构域组成的抗体片段；术语“Fab”意指由VL、VH、CL和CH1结构域组成的抗体片段；术语“F(ab')2”意指包含通过铰链区上的二硫桥连接的两个Fab片段的抗体片段。
本文所用的术语“可变”表示抗体中可变区的某些部分在序列上有所不同，它形成了各种特定抗体对其特定抗原的结合和特异性。可变性集中于轻链和重链可变区中称为互补决定区(CDR)或超变区中的三个片段中。天然重链和轻链的可变区中各自包含四个FR区(可变区中较保守的部分)，它们大致上呈β-折叠构型，由形成连接环的三个CDR相连，可形成部分β折叠结构。每条链中的CDR通过FR区紧密地靠在一起并与另一链的CDR一起形成了抗体的抗原结合部位。恒定区不直接参与抗体与抗原的结合，但是它们表现出不同的效应功能，例如参与抗体的依赖于抗体的细胞毒性。
本发明的抗体可以是游离形式或缀合形式。缀合形式是与能够调节体内持续性、促进或限制体内分布、降低对蛋白水解剂的敏感性、降低抗原性、提高细胞毒能力和/或易于在体液和组织中检测的分子缀合的上述的抗体。适于缀合的分子的非限制例子包括人血清白蛋白，麦芽糖结合蛋白，谷胱甘肽-S-转移酶，噬菌体外壳p3或p8蛋白，肽，糖，PEG或PEG样分子，动物、植物或微生物源毒素，细胞因子，酶，化学发光化合物，生物发光化合物，金属原子，放射性同位素，荧光化合物，标签基团或磷酸化、糖基化、泛素化、苏素化或内肽酶解的底物。为便于缀合，抗体的C末端或N末端可以是修饰的，例如，通过插入额外的氨基酸残基，如一个或更多个可形成二硫桥的半胱氨酸残基。本发明应用的抗体还可以连接人红血球或其它细胞载体，连接特定制剂，或连接缓释系统例如但不限于脂质体、树形聚合物、微粒体、纳米颗粒、微胶囊、病毒载体等等。
本发明还提供了一种分离的核酸分子，所述核酸分子编码SEQ ID NO:1所示的氨基酸序列或SEQ IDNO:5所示的氨基酸序列。
本发明的编码前面所述的抗体或其抗原结合片段的核酸分子包括具有上述核苷酸序列的保守核苷酸序列变体的核酸分子。所谓的保守核苷酸序列变体源于遗传密码简并和沉默的变体，核苷酸的替代、缺失和增加也包含在内。
本发明还提供了一种前面所述的核酸分子的DNA片段。所述DNA片段可以编码抗体重链或轻链可变区的任何区域，包括重链CDR1、CDR2、或CDR3，或轻链CDR1、CDR2、或CDR3。
本发明的另一目标是治疗或预防HCV感染的药物组合物，包含药学有效量的上述定义的单克隆抗体或其片段。
所述药物组合物还包括药学上可接受的载体，所述载体包括但不限于已经被美国食品与药品管理局认可的而可用于人类或动物的任何佐剂、载体、赋形剂、助流剂、稀释剂、表面活性剂、润湿剂、分散剂、助悬剂、稳定剂、等渗压剂、溶剂或乳化剂等对组成药物组合物无副作用的各种形式的载体。
本发明的组合物可被施用给被HCV感染的或有感染风险的受试者。可以应用任何合适的施用途径，包括胃肠外的、口服、眼、局部(topical)、局部区域(loco-regional)、灌肠或气溶胶给药。胃肠外给药包括肌内注射、静脉注射、淋巴管内注射、皮下或皮内注射及输注。
本发明还提供了一种前面所述的全人源抗HCV的单克隆中和抗体的制备方法，所述制备方法采用的是单个B淋巴细胞抗体制备技术。所述单个B淋巴细胞抗体制备技术是将单细胞分离鉴定技术结合多种PCR技术形成的一种单克隆抗体的体外表达系统，即从人的B细胞中直接获取全人源的单克隆抗体的方法。
进一步，所述方法包括如下步骤：
(1)检测发现HCV感染患者；
(2)分离HCV感染患者静脉血中的单个核细胞，之后利用流式细胞术进行细胞分选，分选出含有CD235a-/IgD-/CD20+/CD27ALL的记忆性B细胞；
(3)利用单细胞RT-PCR扩增步骤(2)获得的单个B细胞中的抗体轻链和重链可变区的核苷酸片段；
(4)将步骤(3)获得的抗体轻链和重链可变区的核苷酸片段融合到含有人类抗体恒定区的表达载体中形成重组表达载体，之后导入宿主细胞表达；
(5)抗体筛选平台筛选获得具有结合活性和中和活性的本发明的全人源抗HCV的单克隆中和抗体。
本发明还提供了前面所述的全人源抗HCV的单克隆中和抗体或其片段在制备HCV检测产品中的应用。
所述检测产品包括但不限于检测试剂、试剂盒、芯片或试纸。凡是包括前面所述结合分子的能够检测出HCV的检测产品均包括在本发明的范围之内。
本发明还提供了前面所述的全人源抗HCV的单克隆中和抗体或其片段在制备前面所述的药物组合物中的应用。
本发明还提供了前面所述的全人源抗HCV的单克隆中和抗体或其片段在制备抑制HCV药物中的应用。
本发明还提供了前面所述的全人源抗HCV的单克隆中和抗体或其抗原结合片段在制备检测、预防或治疗由HCV感染引起的疾病的产品中的应用。
进一步，所述由HCV感染引起的疾病包括肝病，如肝炎、肝硬化或肝癌和肝外疾病。
本发明的肝炎是指急性或慢性丙型肝炎。
本发明的抗体TRN1007可以用于治疗急性、慢性丙型肝炎患者，所述治疗是通过给予这些患者治疗有效量的能够结合HCV E2的抗体。“治疗有效量”是指能够在个体中有效减轻HCV感染症状的剂量，或者是有效降低循环中病毒颗粒的剂量。该抗体也可以用于例如HCV阳性携带者分娩的新生儿的被动免疫，还可以用于肝移植的被动免疫以防止这些患者可能出现的反复HCV感染。特别地，对于预计没有来自干扰素治疗的治疗效果的、感染基因型的丙型肝炎患者，可以减轻不良反应，并且可以提供选择新的治疗方法的机会。
附图说明
图1为抗体TRN1007的SDS-PAGE和Western-blot检测图；左侧为SDS-PAGE图，1为非还原样品，M为marker，2为还原样品；右侧为Western-blot图，1为非还原样品，M为marker，2为还原样品；
图2是抗体与不同亚型的HCV病毒中和活性测定图；
图3是抗体与HCV7种不同基因型的抗原结合检测图；
图4是BiaCore检测抗体TRN1007与4种不同基因型抗原的亲和力结果图；a为E2-core-2a、b为E2-core-4a、c为E2-core-6a、d为E2-core-7a。
具体实施方式
下面通过实施例进一步说明本发明。应该理解的是，本发明的实施例是用于说明本发明而不是对本发明的限制。根据本发明的实质对本发明进行的简单改进都属于本发明要求保护的范围。
实施例
1、记忆性B细胞分选策略
流式细胞术(Flow CytoMetry，FCM)是高速、逐一的细胞定量分析和分选的技术，代表了一种用于选择高亲和力人单克隆抗体的高度有效工具。实际上，基于流式技术的分选步骤非常灵活，并且可被优化用以选择交叉反应的抗体。特别地，HCV-I002血液样本源于已确认的HCV感染患者，淋巴细胞分离管分离外周血单个核细胞(Peripheral bloodmononuclear cell，PBMC)后利用BD FACSria流式细胞仪(BD Biosciences,San Jose，CA)进行细胞分选，获得特异性标记荧光素的抗原得到E2双荧光标记的靶细胞。简要地，用这种方法可以获得在自然感染过程中产生的抗体，但其仍然能够结合从未被所选择研究患者的免疫系统遇到过的不同糖蛋白。
2、重链和轻链序列获取
将含有单个B淋巴细胞进行逆转录PCR条件，产物cDNA采用巢式PCR方法扩增重链和轻链的可变区目的基因。使用2％琼脂糖凝胶进行电泳实验进行鉴定，凝胶成像系统检测PCR产物大小。鉴定获得的重链和轻链序列均为全人源化序列。
3、鉴别目的抗体
在大肠杆菌DH5α中大量扩增表达阳性抗体重链和轻链基因的质粒，抽提后用转染试剂Polyetherimide共转染CHO细胞，培养、收集细胞上清进行ELISA筛选检测。将筛选到的有结合活性的抗体进行中和实验，中和效率最好的抗体即为目的抗体TRN1007。
4、抗体大量表达与纯化
将中和实验鉴定出的有中和活性的编号为HCV的抗体重链与轻链的表达载体(其中，重链可变区的氨基酸序列如SEQ ID NO:1所示；抗体轻链可变区的氨基酸序列如SEQ IDNO:5所示)转染CHO细胞，37℃、5％CO2培养箱培养96小时。然后收集转染上清，亲和层析法进行纯化。通过SDS-PAGE和Western-blot检验HCV抗体的表达及纯化结果，结果如图1所示，得到较纯蛋白，并可清晰观察到解链后的抗体轻、重链。
实施例2 TRN1007抗体与不同的HCV真病毒株的中和活性评估
简要地，铺Huh7细胞于96孔板中，过夜培养，感染时细胞约30％铺满。HCV真病毒株(h77、Con1、JFH1、S52、ED43、SA13、HK6a)与不同浓度的抗体混合，起始浓度25μg/ml，3倍稀释，室温放置30min。加入HCV真病毒株(h77、Con1、JFH1、S52、ED43、SA13、HK6a)与抗体混合液于96孔板，感染Huh7细胞。对照组仅加入HCV病毒株2G9，感染24h后换液，继续培养1-2天。感染后2-3天后测定活性，读取荧光值。比较抗体与对照组，计算中和效率。
结果如表1和图2所示。TRN1007抗体对HCV真病毒株JFH1(2a)、S52(3a)、ED43(4a)、HK6a的中和能力均很强，特别是病毒株S52(3a)和HK6a，IC50值仅为4.6nM和6.6nM。中和活性是抗体抗病毒免疫的主要指标之一，TRN1007抗体优异的中和活性显示其在免疫治疗和预防的良好应用前景。
表1 TRN1007抗体与不同的HCV病毒株的中和效率
| 病毒株/mAb(ug/mL) | TRN1007 | 2G9 |
| H77(1a) | 0.3096 | >50 |
| Con1(1b) | 41.00 | >50 |
| JFH1(2a) | 0.02666 | >50 |
| S52(3a) | 0.004595 | >50 |
| ED43(4a) | 0.07398 | >50 |
| SA13(5a) | 1.633 | >50 |
| HK6a | 0.006617 | >50 |
实施例3 TRN1007抗体与不同基因型来源的HCV抗原的结合活性评估
以不同的HCV病毒株(1a、1b、2、3、4、5、6、7)膜蛋白E2为抗原，并用包被液将抗原稀释浓度为100ng/ml，包被于ELISA 96孔板，每孔100μl，4℃过夜。封闭液37℃封闭2个小时。封闭后加一抗，起始浓度为25μg/ml，3倍梯度稀释，每孔体积为100μl，37℃孵育1个小时，同时用HCV阳性病人血清作为阳性对照，狂犬抗体为阴性对照。用HRP标记的羊抗人IgG(1:2000稀释)作为二抗37℃孵育1个小时。加入底物显色液(TMB)100μL/孔，37℃避光放置5min后，用2M硫酸中止反应，用450nm波长进行比色。
结果如图3所示，表明TRN1007抗体能够结合所有不同的HCV病毒株(1a、1b、2、3、4、5、6、7)的膜蛋白E2，即显示TRN1007抗体具有识别所有不同的HCV病毒；其相对于用对照HCV阳性病人血清具有更高的OD值，而且低浓度TRN1007抗体和不同抗原的结合活性依然很强。
实施例4 TRN1007抗体与HCV的包膜糖蛋白的亲和活性评估
利用捕获法将羊抗人IgG抗体作为捕获分子固定在CM5传感器芯片上，按照偶联试剂盒的操作将其偶联到CM5芯片金薄膜表面上，然后活化活化芯片的葡聚糖表面、确定偶联量，乙醇胺封闭表面残留的活化基团；CM5芯片上捕获分子捕获配体：将制备的TRN1007抗体作为配体。TRN1007与抗原即不同基因型的HCV的包膜糖蛋白(HCV-E2-core)结合的亲和力和动力学分析：HCV-E2-core蛋白用HBS-EP缓冲液稀释作为分析物，分析物以逐渐增高的浓度(20μg/ml、40μg/ml、80μg/ml和160μg/ml)依次流过芯片，分别得到信号曲线(见图4)。每个浓度作为1个循环，完成1次循环后用3mol/L的氯化镁再生芯片以回复到原始未结合抗原的状态。
用BiaCore X-100System软件进行分析，进而得到亲和力和动力学参数，具体见表2，TRN1007抗体与抗原E2-core-2a、E2-core-4a、E2-core-6a和E2-core-7a的KD值分别为1.07E-12M、1.59E-12M、1.79E-10M、1.78E-11M，显示抗体与不同的抗原亲和力都较高。
表2 BiaCore X-100测定亲和力和动力学结果参数表
| Ligand | ka(1/Ms) | kd(1/s) | KD(M) |
| E2-core-2a | 8.05E+04 | 8.63E-08 | 1.07E-12 |
| E2-core-4a | 4.51E+04 | 7.17E-08 | 1.59E-12 |
| E2-core-6a | 2994 | 5.36E-07 | 1.79E-10 |
| E2-core-7a | 9.44E+04 | 1.68E-06 | 1.78E-11 |
虽然以上仅描述了本发明的具体实施方式范例，但是本领域的技术人员应当理解，这些仅是举例说明，本发明的保护范围是由所附权利要求书限定的。本领域的技术人员在不背离本发明的原理和实质的前提下，可以对这些实施方式做出多种变更或修改，但这些变更或修改均落入本发明的保护范围。
SEQUENCE LISTING
<110> 广州泰诺迪生物科技有限公司
珠海泰诺麦博生物技术有限公司
暨南大学
<120> 一种抗丙型肝炎病毒抗体的获取以及应用
<160> 8
<170> PatentIn version 3.3
<210> 1
<211> 128
<212> PRT
<213> 人工序列
<400> 1
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Ser Phe Arg Ser Tyr
20 25 30
Gly Phe Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Arg Ile Ile Pro Ala Ala Asn Phe Ala Asp Tyr Ala Pro Lys Phe
50 55 60
Gln Asp Arg Val Thr Ile Ser Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Lys Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gln Asp Leu Ser Gly Glu Leu Thr Ala Leu Tyr Tyr Tyr
100 105 110
His Gly Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120 125
<210> 2
<211> 8
<212> PRT
<213> 人工序列
<400> 2
Gly Gly Ser Phe Arg Ser Tyr Gly
1 5
<210> 3
<211> 8
<212> PRT
<213> 人工序列
<400> 3
Ile Ile Pro Ala Ala Asn Phe Ala
1 5
<210> 4
<211> 21
<212> PRT
<213> 人工序列
<400> 4
Ala Arg Asp Gln Asp Leu Ser Gly Glu Leu Thr Ala Leu Tyr Tyr Tyr
1 5 10 15
His Gly Leu Asp Val
20
<210> 5
<211> 108
<212> PRT
<213> 人工序列
<400> 5
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Gln Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Asn
20 25 30
Tyr Ile Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Ser Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Thr Gly Ser Gly Thr Asp Phe Thr Leu Ala Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Asp Ser Phe
85 90 95
Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 6
<211> 7
<212> PRT
<213> 人工序列
<400> 6
Gln Ser Val Ser Asn Asn Tyr
1 5
<210> 7
<211> 3
<212> PRT
<213> 人工序列
<400> 7
Gly Ala Ser
1
<210> 8
<211> 9
<212> PRT
<213> 人工序列
<400> 8
Gln Gln Tyr Gly Asp Ser Phe Trp Thr
1 5
Claims (16)
1.一种能够结合来自多种不同基因型的HCV病毒的全人源单克隆中和抗体或其抗原结合片段，其特征在于，抗体包括重链可变区的互补性决定区域CDR1、CDR2、CDR3和轻链可变区的互补性决定区域CDR1、CDR2、CDR3；重链CDR1、CDR2、CDR3的氨基酸序列分别如SEQ IDNO:2、SEQ ID NO:3、SEQ ID NO:4所示；轻链CDR1、CDR2、CDR3的氨基酸序列分别如SEQ IDNO:6、SEQ ID NO:7、SEQ ID NO:8所示；抗原结合片段选自Fab、Fab′、F(ab′)2、Fv、dAb和互补性决定区片段、单链抗体、嵌合抗体、双抗，或选自来源于免疫球蛋白重链可变区与轻链可变区的CDR片段的一个或更多个拷贝的骨架。
2.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，抗体的重链可变区具有SEQ ID NO:1所示的氨基酸序列；抗体的轻链可变区具有SEQ IDNO:5所示的氨基酸序列。
3.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，抗原结合片段为同时包含重链与轻链可变区或CDRs的单链抗体。
4.分离的核酸分子，其特征在于，核酸分子编码权利要求1-3中任一项所述的抗体或其抗原结合片段。
5.一种包括权利要求4所述的核酸分子的表达载体。
6.根据权利要求5所述的表达载体，其特征在于，原核表达载体为PET28a、pGEM Ex1、pGEM7ZF(+)或pBAD24，真核表达载体为pBudCE4.1、pcDNA3.1、pCDNA3.3、pCMV-Tag2B或pGL3basic。
7.一种包括权利要求4所述的核酸分子或权利要求5所述的表达载体的宿主细胞。
8.根据权利要求7所述的宿主细胞，其特征在于，宿主细胞为CHO，COS，293细胞或Bowes黑素瘤细胞。
9.一种包括权利要求1-3中任一项所述的抗体或其抗原结合片段的药物组合物。
10.根据权利要求9所述的药物组合物，其特征在于，药物组合物还包括治疗HCV的市售药物。
11.一种包括权利要求1-3中任一项所述的抗体或其抗原结合片段的HCV检测产品。
12.权利要求1-3中任一项所述的抗体或其抗原结合片段在制备检测、预防或治疗由HCV感染引起的疾病的产品中的应用。
13.根据权利要求12所述的应用，其特征在于，疾病包括肝病。
14.根据权利要求13所述的应用，其特征在于，肝病为肝炎、肝硬化或肝癌。
15.根据权利要求12所述的应用，其特征在于，疾病包括肝外疾病。
16.根据权利要求14所述的应用，其特征在于，肝炎是急性或慢性丙型肝炎。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710668058.7A CN107286237B (zh) | 2017-08-07 | 2017-08-07 | 一种抗丙型肝炎病毒抗体的获取以及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710668058.7A CN107286237B (zh) | 2017-08-07 | 2017-08-07 | 一种抗丙型肝炎病毒抗体的获取以及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107286237A CN107286237A (zh) | 2017-10-24 |
| CN107286237B true CN107286237B (zh) | 2020-04-03 |
Family
ID=60105326
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710668058.7A Active CN107286237B (zh) | 2017-08-07 | 2017-08-07 | 一种抗丙型肝炎病毒抗体的获取以及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107286237B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115124617B (zh) * | 2021-05-08 | 2024-02-09 | 南昌大学 | 一种高亲和力抗丙型肝炎病毒的全人源单克隆抗体及其用途 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| YU48038B (sh) * | 1987-11-18 | 1996-10-18 | Chiron Corp. | Postupak za dijagnostiku ne-a, ne-v hepatitisa |
| CN1044384C (zh) * | 1994-08-25 | 1999-07-28 | 中国药品生物制品检定所 | 抗丙型肝炎病毒抗原单克隆抗体细胞株建立及其单克隆抗体 |
-
2017
- 2017-08-07 CN CN201710668058.7A patent/CN107286237B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN107286237A (zh) | 2017-10-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106211773B (zh) | 用于登革病毒的抗体分子及其应用 | |
| JP7235256B2 (ja) | 重症熱性血小板減少症候群ウイルスの外膜糖タンパク質に結合する抗体及びその用途 | |
| US7507408B2 (en) | Anti hepatitis C virus antibody and uses thereof | |
| US7727529B2 (en) | Human monoclonal antibody Fab fragments directed against HCV E2 glycoprotein and endowed with in vitro neutralizing activity | |
| CN111420048B (zh) | 抗basigin人源化抗体用于制备治疗新型冠状病毒肺炎药物的应用 | |
| JP2019509311A (ja) | デングウイルスに対する抗体分子の製剤 | |
| CN102264393B (zh) | 作为治疗和预防hcv感染药物的抗hcv单克隆抗体 | |
| EP4206224A1 (en) | Human antibody or antigen-binding fragment thereof against coronavirus spike protein | |
| WO2019128119A1 (zh) | 一种针对破伤风毒素的全人源单克隆中和抗体及其应用 | |
| WO2012155819A1 (zh) | Hbv特异性抗体 | |
| CN107286237B (zh) | 一种抗丙型肝炎病毒抗体的获取以及应用 | |
| CN106749645B (zh) | 一种全人源抗丙型肝炎病毒的中和抗体 | |
| US10888615B2 (en) | Neutralizing human monoclonal antibody 8D6 against HCV infection | |
| US20230348568A1 (en) | Epstein-barr virus monoclonal antibodies and uses thereof | |
| EP4282880A1 (en) | Fully human broad-spectrum neutralizing antibody 76e1 against coronavirus, and use thereof | |
| WO2021142672A1 (zh) | 抗Echo30的抗体，相应的核酸，载体，宿主细胞，组合物 | |
| KR20220122467A (ko) | 사스-코로나바이러스-2에 특이적으로 결합하는 항체 및 그의 용도 | |
| WO2024054822A1 (en) | Engineered sars-cov-2 antibodies with increased neutralization breadth |
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 | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 510300 unit 207, second floor, office area, No. 1, helix 4th Road, International Biological Island, Guangzhou, Guangdong Patentee after: GUANGZHOU TAINUODI BIOTECHNOLOGY Co.,Ltd. Patentee after: Zhuhai Tainuo Maibo Pharmaceutical Co.,Ltd. Patentee after: Jinan University Address before: 510300 unit 207, second floor, office area, No. 1, helix 4th Road, International Biological Island, Guangzhou, Guangdong Patentee before: GUANGZHOU TAINUODI BIOTECHNOLOGY Co.,Ltd. Patentee before: ZHUHAI TRINOMAB BIOTECHNOLOGY Co.,Ltd. Patentee before: Jinan University |
|
| CP01 | Change in the name or title of a patent holder | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230828 Address after: 519090 room 110, first floor, plant 6, No. 6366, Zhuhai Avenue, Jinwan District, Zhuhai City, Guangdong Province Patentee after: Zhuhai Tainuo Maibo Pharmaceutical Co.,Ltd. Patentee after: Jinan University Address before: 510300 unit 207, second floor, office area, No. 1, helix 4th Road, International Biological Island, Guangzhou, Guangdong Patentee before: GUANGZHOU TAINUODI BIOTECHNOLOGY Co.,Ltd. Patentee before: Zhuhai Tainuo Maibo Pharmaceutical Co.,Ltd. Patentee before: Jinan University |
|
| TR01 | Transfer of patent right |