CN115028712A - Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof - Google Patents
Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof Download PDFInfo
- Publication number
- CN115028712A CN115028712A CN202210682852.8A CN202210682852A CN115028712A CN 115028712 A CN115028712 A CN 115028712A CN 202210682852 A CN202210682852 A CN 202210682852A CN 115028712 A CN115028712 A CN 115028712A
- Authority
- CN
- China
- Prior art keywords
- variable region
- chain variable
- ser
- monoclonal antibody
- antibody
- 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
- 241000711549 Hepacivirus C Species 0.000 title claims abstract description 47
- 239000012634 fragment Substances 0.000 claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 11
- 239000013604 expression vector Substances 0.000 claims abstract description 10
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 4
- 239000002773 nucleotide Substances 0.000 claims description 22
- 125000003729 nucleotide group Chemical group 0.000 claims description 22
- 101710125507 Integrase/recombinase Proteins 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
- 239000003937 drug carrier Substances 0.000 claims description 4
- 208000010710 hepatitis C virus infection Diseases 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 54
- 208000005176 Hepatitis C Diseases 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 8
- 241000700605 Viruses Species 0.000 abstract description 7
- 238000011282 treatment Methods 0.000 abstract description 7
- 229940079593 drug Drugs 0.000 abstract description 5
- 230000005847 immunogenicity Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 241001465754 Metazoa Species 0.000 abstract description 3
- 241001529936 Murinae Species 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 63
- 150000001413 amino acids Chemical class 0.000 description 29
- 238000000034 method Methods 0.000 description 29
- 239000013612 plasmid Substances 0.000 description 24
- 210000001806 memory b lymphocyte Anatomy 0.000 description 20
- 102000004169 proteins and genes Human genes 0.000 description 18
- 239000000243 solution Substances 0.000 description 18
- 239000000872 buffer Substances 0.000 description 15
- 238000012216 screening Methods 0.000 description 15
- 239000000523 sample Substances 0.000 description 12
- 238000005406 washing Methods 0.000 description 12
- 241000283707 Capra Species 0.000 description 11
- 102000036639 antigens Human genes 0.000 description 10
- 108091007433 antigens Proteins 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 10
- 239000008280 blood Substances 0.000 description 10
- 108020004414 DNA Proteins 0.000 description 9
- HQYVQDRYODWONX-DCAQKATOSA-N Val-His-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CO)C(=O)O)N HQYVQDRYODWONX-DCAQKATOSA-N 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 9
- 238000001514 detection method Methods 0.000 description 9
- 239000000427 antigen Substances 0.000 description 8
- 238000007857 nested PCR Methods 0.000 description 8
- 239000013598 vector Substances 0.000 description 8
- 238000004113 cell culture Methods 0.000 description 7
- 239000012228 culture supernatant Substances 0.000 description 7
- 238000001976 enzyme digestion Methods 0.000 description 7
- 238000000338 in vitro Methods 0.000 description 7
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 6
- 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 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 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 6
- 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 6
- 108010081404 acein-2 Proteins 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 6
- 210000004180 plasmocyte Anatomy 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 238000012795 verification Methods 0.000 description 6
- SOEGEPHNZOISMT-BYPYZUCNSA-N Gly-Ser-Gly Chemical compound NCC(=O)N[C@@H](CO)C(=O)NCC(O)=O SOEGEPHNZOISMT-BYPYZUCNSA-N 0.000 description 5
- 241000880493 Leptailurus serval Species 0.000 description 5
- JGUWRQWULDWNCM-FXQIFTODSA-N Ser-Val-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O JGUWRQWULDWNCM-FXQIFTODSA-N 0.000 description 5
- 230000003321 amplification Effects 0.000 description 5
- 210000003719 b-lymphocyte Anatomy 0.000 description 5
- 230000029087 digestion Effects 0.000 description 5
- 238000003199 nucleic acid amplification method Methods 0.000 description 5
- 239000008188 pellet Substances 0.000 description 5
- 238000011084 recovery Methods 0.000 description 5
- 239000012096 transfection reagent Substances 0.000 description 5
- GYWQGGUCMDCUJE-DLOVCJGASA-N Asp-Phe-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(O)=O GYWQGGUCMDCUJE-DLOVCJGASA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- BPHKULHWEIUDOB-FXQIFTODSA-N Cys-Gln-Gln Chemical compound SC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O BPHKULHWEIUDOB-FXQIFTODSA-N 0.000 description 4
- 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 4
- RWQCWSGOOOEGPB-FXQIFTODSA-N Gln-Ser-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O RWQCWSGOOOEGPB-FXQIFTODSA-N 0.000 description 4
- ITVBKCZZLJUUHI-HTUGSXCWSA-N Glu-Phe-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ITVBKCZZLJUUHI-HTUGSXCWSA-N 0.000 description 4
- 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 4
- ADJDNJCSPNFFPI-FXQIFTODSA-N Ser-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO ADJDNJCSPNFFPI-FXQIFTODSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- YOPQYBJJNSIQGZ-JNPHEJMOSA-N Thr-Tyr-Tyr Chemical compound C([C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 YOPQYBJJNSIQGZ-JNPHEJMOSA-N 0.000 description 4
- GBIUHAYJGWVNLN-UHFFFAOYSA-N Val-Ser-Pro Natural products CC(C)C(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O GBIUHAYJGWVNLN-UHFFFAOYSA-N 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 108010089804 glycyl-threonine Proteins 0.000 description 4
- 108010010147 glycylglutamine Proteins 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 238000010839 reverse transcription Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 230000003612 virological effect Effects 0.000 description 4
- QDGMZAOSMNGBLP-MRFFXTKBSA-N Ala-Trp-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)N QDGMZAOSMNGBLP-MRFFXTKBSA-N 0.000 description 3
- 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 3
- OBFTYSPXDRROQO-SRVKXCTJSA-N Arg-Gln-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCCN=C(N)N OBFTYSPXDRROQO-SRVKXCTJSA-N 0.000 description 3
- AOHKLEBWKMKITA-IHRRRGAJSA-N Arg-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AOHKLEBWKMKITA-IHRRRGAJSA-N 0.000 description 3
- 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 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- AVZHGSCDKIQZPQ-CIUDSAMLSA-N Glu-Arg-Ala Chemical compound C[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O AVZHGSCDKIQZPQ-CIUDSAMLSA-N 0.000 description 3
- 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 3
- 108010065920 Insulin Lispro Proteins 0.000 description 3
- NRFGTHFONZYFNY-MGHWNKPDSA-N Leu-Ile-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NRFGTHFONZYFNY-MGHWNKPDSA-N 0.000 description 3
- 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 3
- LJBVRCDPWOJOEK-PPCPHDFISA-N Leu-Thr-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LJBVRCDPWOJOEK-PPCPHDFISA-N 0.000 description 3
- 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 3
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 3
- 101710144111 Non-structural protein 3 Proteins 0.000 description 3
- 108700026244 Open Reading Frames Proteins 0.000 description 3
- 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 3
- JMVQDLDPDBXAAX-YUMQZZPRSA-N Pro-Gly-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 JMVQDLDPDBXAAX-YUMQZZPRSA-N 0.000 description 3
- RTQKBZIRDWZLDF-BZSNNMDCSA-N Pro-Pro-Trp Chemical compound C([C@H]1C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)O)CCN1C(=O)[C@@H]1CCCN1 RTQKBZIRDWZLDF-BZSNNMDCSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 101710185720 Putative ethidium bromide resistance protein Proteins 0.000 description 3
- 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 3
- FIDMVVBUOCMMJG-CIUDSAMLSA-N Ser-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO FIDMVVBUOCMMJG-CIUDSAMLSA-N 0.000 description 3
- 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 3
- 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 3
- 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 3
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 3
- CAGTXGDOIFXLPC-KZVJFYERSA-N Thr-Arg-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CCCN=C(N)N CAGTXGDOIFXLPC-KZVJFYERSA-N 0.000 description 3
- IMULJHHGAUZZFE-MBLNEYKQSA-N Thr-Gly-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IMULJHHGAUZZFE-MBLNEYKQSA-N 0.000 description 3
- BIBYEFRASCNLAA-CDMKHQONSA-N Thr-Phe-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 BIBYEFRASCNLAA-CDMKHQONSA-N 0.000 description 3
- CNLKDWSAORJEMW-KWQFWETISA-N Tyr-Gly-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](C)C(O)=O CNLKDWSAORJEMW-KWQFWETISA-N 0.000 description 3
- PZTZYZUTCPZWJH-FXQIFTODSA-N Val-Ser-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PZTZYZUTCPZWJH-FXQIFTODSA-N 0.000 description 3
- 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 3
- 230000004913 activation Effects 0.000 description 3
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 3
- 108010087924 alanylproline Proteins 0.000 description 3
- 239000011324 bead Substances 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 238000010276 construction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 238000012917 library technology Methods 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 108010017391 lysylvaline Proteins 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 229940125645 monoclonal antibody drug Drugs 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- OMDNCNKNEGFOMM-BQBZGAKWSA-N Ala-Met-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O OMDNCNKNEGFOMM-BQBZGAKWSA-N 0.000 description 2
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 2
- 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 2
- VYZBPPBKFCHCIS-WPRPVWTQSA-N Arg-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N VYZBPPBKFCHCIS-WPRPVWTQSA-N 0.000 description 2
- JRVABKHPWDRUJF-UBHSHLNASA-N Asn-Asn-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N JRVABKHPWDRUJF-UBHSHLNASA-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
- ANRZCQXIXGDXLR-CWRNSKLLSA-N Asn-Trp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CC(=O)N)N)C(=O)O ANRZCQXIXGDXLR-CWRNSKLLSA-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
- YFXFOZPXVFPBDH-VZFHVOOUSA-N Cys-Ala-Thr Chemical compound C[C@@H](O)[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)CS)C(O)=O YFXFOZPXVFPBDH-VZFHVOOUSA-N 0.000 description 2
- 101710118188 DNA-binding protein HU-alpha Proteins 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 102100038132 Endogenous retrovirus group K member 6 Pro protein Human genes 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- WPJDPEOQUIXXOY-AVGNSLFASA-N Gln-Tyr-Asn Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O WPJDPEOQUIXXOY-AVGNSLFASA-N 0.000 description 2
- MXOODARRORARSU-ACZMJKKPSA-N Glu-Ala-Ser Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)O)N MXOODARRORARSU-ACZMJKKPSA-N 0.000 description 2
- 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 2
- 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 2
- 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 2
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 2
- JYGYNWYVKXENNE-OALUTQOASA-N Gly-Tyr-Trp Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O JYGYNWYVKXENNE-OALUTQOASA-N 0.000 description 2
- WCHONUZTYDQMBY-PYJNHQTQSA-N His-Pro-Ile Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WCHONUZTYDQMBY-PYJNHQTQSA-N 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
- 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
- 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
- 206010067125 Liver injury Diseases 0.000 description 2
- 101710144128 Non-structural protein 2 Proteins 0.000 description 2
- 101800001014 Non-structural protein 5A Proteins 0.000 description 2
- 101710199667 Nuclear export protein Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 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
- VXCHGLYSIOOZIS-GUBZILKMSA-N Pro-Ala-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 VXCHGLYSIOOZIS-GUBZILKMSA-N 0.000 description 2
- VYWNORHENYEQDW-YUMQZZPRSA-N Pro-Gly-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 VYWNORHENYEQDW-YUMQZZPRSA-N 0.000 description 2
- HAAQQNHQZBOWFO-LURJTMIESA-N Pro-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H]1CCCN1 HAAQQNHQZBOWFO-LURJTMIESA-N 0.000 description 2
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 2
- SUENWIFTSTWUKD-AVGNSLFASA-N Pro-Leu-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O SUENWIFTSTWUKD-AVGNSLFASA-N 0.000 description 2
- VBZXFFYOBDLLFE-HSHDSVGOSA-N Pro-Trp-Thr Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H]([C@H](O)C)C(O)=O)C(=O)[C@@H]1CCCN1 VBZXFFYOBDLLFE-HSHDSVGOSA-N 0.000 description 2
- ICHZYBVODUVUKN-SRVKXCTJSA-N Ser-Asn-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ICHZYBVODUVUKN-SRVKXCTJSA-N 0.000 description 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
- JIPVNVNKXJLFJF-BJDJZHNGSA-N Ser-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N JIPVNVNKXJLFJF-BJDJZHNGSA-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
- WLJPJRGQRNCIQS-ZLUOBGJFSA-N Ser-Ser-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O WLJPJRGQRNCIQS-ZLUOBGJFSA-N 0.000 description 2
- 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 2
- 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 2
- KZTLZZQTJMCGIP-ZJDVBMNYSA-N Thr-Val-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KZTLZZQTJMCGIP-ZJDVBMNYSA-N 0.000 description 2
- HKIUVWMZYFBIHG-KKUMJFAQSA-N Tyr-Arg-Gln Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O HKIUVWMZYFBIHG-KKUMJFAQSA-N 0.000 description 2
- PZXUIGWOEWWFQM-SRVKXCTJSA-N Tyr-Asn-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O PZXUIGWOEWWFQM-SRVKXCTJSA-N 0.000 description 2
- ANHVRCNNGJMJNG-BZSNNMDCSA-N Tyr-Tyr-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CS)C(=O)O)N)O ANHVRCNNGJMJNG-BZSNNMDCSA-N 0.000 description 2
- 108091023045 Untranslated Region Proteins 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003443 antiviral agent Substances 0.000 description 2
- 108010008355 arginyl-glutamine Proteins 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 238000010370 cell cloning Methods 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 231100000753 hepatic injury Toxicity 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 231100000283 hepatitis Toxicity 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- 108010064235 lysylglycine Proteins 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000011325 microbead Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 210000005087 mononuclear cell Anatomy 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 108010051242 phenylalanylserine Proteins 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 239000003161 ribonuclease inhibitor Substances 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 108010084932 tryptophyl-proline Proteins 0.000 description 2
- 108010038745 tryptophylglycine Proteins 0.000 description 2
- 108010044292 tryptophyltyrosine Proteins 0.000 description 2
- LNAZSHAWQACDHT-XIYTZBAFSA-N (2r,3r,4s,5r,6s)-4,5-dimethoxy-2-(methoxymethyl)-3-[(2s,3r,4s,5r,6r)-3,4,5-trimethoxy-6-(methoxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6r)-4,5,6-trimethoxy-2-(methoxymethyl)oxan-3-yl]oxyoxane Chemical compound CO[C@@H]1[C@@H](OC)[C@H](OC)[C@@H](COC)O[C@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@H](O[C@H]2[C@@H]([C@@H](OC)[C@H](OC)O[C@@H]2COC)OC)O[C@@H]1COC LNAZSHAWQACDHT-XIYTZBAFSA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- UZOVYGYOLBIAJR-UHFFFAOYSA-N 4-isocyanato-4'-methyldiphenylmethane Chemical compound C1=CC(C)=CC=C1CC1=CC=C(N=C=O)C=C1 UZOVYGYOLBIAJR-UHFFFAOYSA-N 0.000 description 1
- YWWATNIVMOCSAV-UBHSHLNASA-N Ala-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YWWATNIVMOCSAV-UBHSHLNASA-N 0.000 description 1
- BUDNAJYVCUHLSV-ZLUOBGJFSA-N Ala-Asp-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O BUDNAJYVCUHLSV-ZLUOBGJFSA-N 0.000 description 1
- MAZZQZWCCYJQGZ-GUBZILKMSA-N Ala-Pro-Arg Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MAZZQZWCCYJQGZ-GUBZILKMSA-N 0.000 description 1
- DCVYRWFAMZFSDA-ZLUOBGJFSA-N Ala-Ser-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DCVYRWFAMZFSDA-ZLUOBGJFSA-N 0.000 description 1
- VJVQKGYHIZPSNS-FXQIFTODSA-N Ala-Ser-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N VJVQKGYHIZPSNS-FXQIFTODSA-N 0.000 description 1
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 1
- HCBKAOZYACJUEF-XQXXSGGOSA-N Ala-Thr-Gln Chemical compound N[C@@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCC(N)=O)C(=O)O HCBKAOZYACJUEF-XQXXSGGOSA-N 0.000 description 1
- LSMDIAAALJJLRO-XQXXSGGOSA-N Ala-Thr-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O LSMDIAAALJJLRO-XQXXSGGOSA-N 0.000 description 1
- AAWLEICNDUHIJM-MBLNEYKQSA-N Ala-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C)N)O AAWLEICNDUHIJM-MBLNEYKQSA-N 0.000 description 1
- VNFSAYFQLXPHPY-CIQUZCHMSA-N Ala-Thr-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNFSAYFQLXPHPY-CIQUZCHMSA-N 0.000 description 1
- IOFVWPYSRSCWHI-JXUBOQSCSA-N Ala-Thr-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C)N IOFVWPYSRSCWHI-JXUBOQSCSA-N 0.000 description 1
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 1
- 108010082126 Alanine transaminase Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- GIVATXIGCXFQQA-FXQIFTODSA-N Arg-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N GIVATXIGCXFQQA-FXQIFTODSA-N 0.000 description 1
- PQWTZSNVWSOFFK-FXQIFTODSA-N Arg-Asp-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)CN=C(N)N PQWTZSNVWSOFFK-FXQIFTODSA-N 0.000 description 1
- YHQGEARSFILVHL-HJGDQZAQSA-N Arg-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N)O YHQGEARSFILVHL-HJGDQZAQSA-N 0.000 description 1
- LVMUGODRNHFGRA-AVGNSLFASA-N Arg-Leu-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O LVMUGODRNHFGRA-AVGNSLFASA-N 0.000 description 1
- JEXPNDORFYHJTM-IHRRRGAJSA-N Arg-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCN=C(N)N JEXPNDORFYHJTM-IHRRRGAJSA-N 0.000 description 1
- COXMUHNBYCVVRG-DCAQKATOSA-N Arg-Leu-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O COXMUHNBYCVVRG-DCAQKATOSA-N 0.000 description 1
- NPAVRDPEFVKELR-DCAQKATOSA-N Arg-Lys-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O NPAVRDPEFVKELR-DCAQKATOSA-N 0.000 description 1
- NIELFHOLFTUZME-HJWJTTGWSA-N Arg-Phe-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NIELFHOLFTUZME-HJWJTTGWSA-N 0.000 description 1
- MFFOYNGMOYFPBD-DCAQKATOSA-N Asn-Arg-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O MFFOYNGMOYFPBD-DCAQKATOSA-N 0.000 description 1
- PNHQRQTVBRDIEF-CIUDSAMLSA-N Asn-Leu-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(=O)N)N PNHQRQTVBRDIEF-CIUDSAMLSA-N 0.000 description 1
- JBDLMLZNDRLDIX-HJGDQZAQSA-N Asn-Thr-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O JBDLMLZNDRLDIX-HJGDQZAQSA-N 0.000 description 1
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 1
- 108010028006 B-Cell Activating Factor Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 101100394003 Butyrivibrio fibrisolvens end1 gene Proteins 0.000 description 1
- 101100268671 Caenorhabditis elegans acc-4 gene Proteins 0.000 description 1
- 101100126625 Caenorhabditis elegans itr-1 gene Proteins 0.000 description 1
- 101710132601 Capsid protein Proteins 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- YUZPQIQWXLRFBW-ACZMJKKPSA-N Cys-Glu-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O YUZPQIQWXLRFBW-ACZMJKKPSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 244000239659 Eucalyptus pulverulenta Species 0.000 description 1
- 108010010803 Gelatin Proteins 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
- IOFDDSNZJDIGPB-GVXVVHGQSA-N Gln-Leu-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IOFDDSNZJDIGPB-GVXVVHGQSA-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
- MSHXWFKYXJTLEZ-CIUDSAMLSA-N Gln-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MSHXWFKYXJTLEZ-CIUDSAMLSA-N 0.000 description 1
- FQCILXROGNOZON-YUMQZZPRSA-N Gln-Pro-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O FQCILXROGNOZON-YUMQZZPRSA-N 0.000 description 1
- KUBFPYIMAGXGBT-ACZMJKKPSA-N Gln-Ser-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O KUBFPYIMAGXGBT-ACZMJKKPSA-N 0.000 description 1
- OSCLNNWLKKIQJM-WDSKDSINSA-N Gln-Ser-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O OSCLNNWLKKIQJM-WDSKDSINSA-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
- VLOLPWWCNKWRNB-LOKLDPHHSA-N Gln-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O VLOLPWWCNKWRNB-LOKLDPHHSA-N 0.000 description 1
- ZFBBMCKQSNJZSN-AUTRQRHGSA-N Gln-Val-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZFBBMCKQSNJZSN-AUTRQRHGSA-N 0.000 description 1
- 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 1
- HGJREIGJLUQBTJ-SZMVWBNQSA-N Glu-Trp-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(C)C)C(O)=O HGJREIGJLUQBTJ-SZMVWBNQSA-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
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- KRRMJKMGWWXWDW-STQMWFEESA-N Gly-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)CN)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KRRMJKMGWWXWDW-STQMWFEESA-N 0.000 description 1
- YZACQYVWLCQWBT-BQBZGAKWSA-N Gly-Cys-Arg Chemical compound [H]NCC(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O YZACQYVWLCQWBT-BQBZGAKWSA-N 0.000 description 1
- DHDOADIPGZTAHT-YUMQZZPRSA-N Gly-Glu-Arg Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DHDOADIPGZTAHT-YUMQZZPRSA-N 0.000 description 1
- XPJBQTCXPJNIFE-ZETCQYMHSA-N Gly-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)CN XPJBQTCXPJNIFE-ZETCQYMHSA-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
- ULZCYBYDTUMHNF-IUCAKERBSA-N Gly-Leu-Glu Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O ULZCYBYDTUMHNF-IUCAKERBSA-N 0.000 description 1
- 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 1
- WNZOCXUOGVYYBJ-CDMKHQONSA-N Gly-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)CN)O WNZOCXUOGVYYBJ-CDMKHQONSA-N 0.000 description 1
- WNGHUXFWEWTKAO-YUMQZZPRSA-N Gly-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN WNGHUXFWEWTKAO-YUMQZZPRSA-N 0.000 description 1
- DBUNZBWUWCIELX-JHEQGTHGSA-N Gly-Thr-Glu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DBUNZBWUWCIELX-JHEQGTHGSA-N 0.000 description 1
- 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
- ZZWUYQXMIFTIIY-WEDXCCLWSA-N Gly-Thr-Leu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O ZZWUYQXMIFTIIY-WEDXCCLWSA-N 0.000 description 1
- MREVELMMFOLESM-HOCLYGCPSA-N Gly-Trp-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C(C)C)C(O)=O MREVELMMFOLESM-HOCLYGCPSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 208000037357 HIV infectious disease Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 208000006083 Hypokinesia Diseases 0.000 description 1
- YOTNPRLPIPHQSB-XUXIUFHCSA-N Ile-Arg-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N YOTNPRLPIPHQSB-XUXIUFHCSA-N 0.000 description 1
- NULSANWBUWLTKN-NAKRPEOUSA-N Ile-Arg-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N NULSANWBUWLTKN-NAKRPEOUSA-N 0.000 description 1
- PFPUFNLHBXKPHY-HTFCKZLJSA-N Ile-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)O)N PFPUFNLHBXKPHY-HTFCKZLJSA-N 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
- PRTZQMBYUZFSFA-XEGUGMAKSA-N Ile-Tyr-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)NCC(=O)O)N PRTZQMBYUZFSFA-XEGUGMAKSA-N 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 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 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 108091026898 Leader sequence (mRNA) Proteins 0.000 description 1
- AXZGZMGRBDQTEY-SRVKXCTJSA-N Leu-Gln-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O AXZGZMGRBDQTEY-SRVKXCTJSA-N 0.000 description 1
- GPICTNQYKHHHTH-GUBZILKMSA-N Leu-Gln-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O GPICTNQYKHHHTH-GUBZILKMSA-N 0.000 description 1
- FEHQLKKBVJHSEC-SZMVWBNQSA-N Leu-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FEHQLKKBVJHSEC-SZMVWBNQSA-N 0.000 description 1
- IRMLZWSRWSGTOP-CIUDSAMLSA-N Leu-Ser-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O IRMLZWSRWSGTOP-CIUDSAMLSA-N 0.000 description 1
- XOWMDXHFSBCAKQ-SRVKXCTJSA-N Leu-Ser-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C XOWMDXHFSBCAKQ-SRVKXCTJSA-N 0.000 description 1
- SVBJIZVVYJYGLA-DCAQKATOSA-N Leu-Ser-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O SVBJIZVVYJYGLA-DCAQKATOSA-N 0.000 description 1
- JGKHAFUAPZCCDU-BZSNNMDCSA-N Leu-Tyr-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CC=C(O)C=C1 JGKHAFUAPZCCDU-BZSNNMDCSA-N 0.000 description 1
- QUCDKEKDPYISNX-HJGDQZAQSA-N Lys-Asn-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QUCDKEKDPYISNX-HJGDQZAQSA-N 0.000 description 1
- GQZMPWBZQALKJO-UWVGGRQHSA-N Lys-Gly-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O GQZMPWBZQALKJO-UWVGGRQHSA-N 0.000 description 1
- 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
- SBQDRNOLGSYHQA-YUMQZZPRSA-N Lys-Ser-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SBQDRNOLGSYHQA-YUMQZZPRSA-N 0.000 description 1
- OHXUUQDOBQKSNB-AVGNSLFASA-N Lys-Val-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O OHXUUQDOBQKSNB-AVGNSLFASA-N 0.000 description 1
- UGCIQUYEJIEHKX-GVXVVHGQSA-N Lys-Val-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O UGCIQUYEJIEHKX-GVXVVHGQSA-N 0.000 description 1
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- SBSIKVMCCJUCBZ-GUBZILKMSA-N Met-Asn-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N SBSIKVMCCJUCBZ-GUBZILKMSA-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
- WYBVBIHNJWOLCJ-UHFFFAOYSA-N N-L-arginyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCCN=C(N)N WYBVBIHNJWOLCJ-UHFFFAOYSA-N 0.000 description 1
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-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
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 101800001020 Non-structural protein 4A Proteins 0.000 description 1
- 101800001019 Non-structural protein 4B Proteins 0.000 description 1
- 239000012124 Opti-MEM Substances 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- GXDPQJUBLBZKDY-IAVJCBSLSA-N Phe-Ile-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GXDPQJUBLBZKDY-IAVJCBSLSA-N 0.000 description 1
- YMIZSYUAZJSOFL-SRVKXCTJSA-N Phe-Ser-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O YMIZSYUAZJSOFL-SRVKXCTJSA-N 0.000 description 1
- 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 1
- BPIMVBKDLSBKIJ-FCLVOEFKSA-N Phe-Thr-Phe Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 BPIMVBKDLSBKIJ-FCLVOEFKSA-N 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 1
- IBGCFJDLCYTKPW-NAKRPEOUSA-N Pro-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]1CCCN1 IBGCFJDLCYTKPW-NAKRPEOUSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102000014961 Protein Precursors Human genes 0.000 description 1
- 108010078762 Protein Precursors Proteins 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 108090000944 RNA Helicases Proteins 0.000 description 1
- 102000004409 RNA Helicases Human genes 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 101800001554 RNA-directed RNA polymerase Proteins 0.000 description 1
- 206010061603 Respiratory syncytial virus infection Diseases 0.000 description 1
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 101100221606 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) COS7 gene Proteins 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- WTWGOQRNRFHFQD-JBDRJPRFSA-N Ser-Ala-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WTWGOQRNRFHFQD-JBDRJPRFSA-N 0.000 description 1
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 1
- IDCKUIWEIZYVSO-WFBYXXMGSA-N Ser-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C)C(O)=O)=CNC2=C1 IDCKUIWEIZYVSO-WFBYXXMGSA-N 0.000 description 1
- BLPYXIXXCFVIIF-FXQIFTODSA-N Ser-Cys-Arg Chemical compound C(C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CO)N)CN=C(N)N BLPYXIXXCFVIIF-FXQIFTODSA-N 0.000 description 1
- HVKMTOIAYDOJPL-NRPADANISA-N Ser-Gln-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O HVKMTOIAYDOJPL-NRPADANISA-N 0.000 description 1
- HJEBZBMOTCQYDN-ACZMJKKPSA-N Ser-Glu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O HJEBZBMOTCQYDN-ACZMJKKPSA-N 0.000 description 1
- 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 1
- JFWDJFULOLKQFY-QWRGUYRKSA-N Ser-Gly-Phe Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JFWDJFULOLKQFY-QWRGUYRKSA-N 0.000 description 1
- GJFYFGOEWLDQGW-GUBZILKMSA-N Ser-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GJFYFGOEWLDQGW-GUBZILKMSA-N 0.000 description 1
- FKYWFUYPVKLJLP-DCAQKATOSA-N Ser-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FKYWFUYPVKLJLP-DCAQKATOSA-N 0.000 description 1
- CUXJENOFJXOSOZ-BIIVOSGPSA-N Ser-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CO)N)C(=O)O CUXJENOFJXOSOZ-BIIVOSGPSA-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
- 108010022999 Serine Proteases Proteins 0.000 description 1
- 102000012479 Serine Proteases Human genes 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- OQCXTUQTKQFDCX-HTUGSXCWSA-N Thr-Glu-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N)O OQCXTUQTKQFDCX-HTUGSXCWSA-N 0.000 description 1
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 1
- IJVNLNRVDUTWDD-MEYUZBJRSA-N Thr-Leu-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IJVNLNRVDUTWDD-MEYUZBJRSA-N 0.000 description 1
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 1
- COYHRQWNJDJCNA-NUJDXYNKSA-N Thr-Thr-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O COYHRQWNJDJCNA-NUJDXYNKSA-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
- SPIFGZFZMVLPHN-UNQGMJICSA-N Thr-Val-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SPIFGZFZMVLPHN-UNQGMJICSA-N 0.000 description 1
- 108091036066 Three prime untranslated region Proteins 0.000 description 1
- 229920001615 Tragacanth Polymers 0.000 description 1
- 108090000340 Transaminases Proteins 0.000 description 1
- RWAYYYOZMHMEGD-XIRDDKMYSA-N Trp-Leu-Ser Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 RWAYYYOZMHMEGD-XIRDDKMYSA-N 0.000 description 1
- QUIXRGCMQOXUSV-SZMVWBNQSA-N Trp-Pro-Pro Chemical compound O=C([C@@H]1CCCN1C(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)N1CCC[C@H]1C(O)=O QUIXRGCMQOXUSV-SZMVWBNQSA-N 0.000 description 1
- HTGJDTPQYFMKNC-VFAJRCTISA-N Trp-Thr-Leu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C(O)=O)[C@@H](C)O)=CNC2=C1 HTGJDTPQYFMKNC-VFAJRCTISA-N 0.000 description 1
- UIDJDMVRDUANDL-BVSLBCMMSA-N Trp-Tyr-Arg 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](CCCNC(N)=N)C(O)=O UIDJDMVRDUANDL-BVSLBCMMSA-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
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 102100036922 Tumor necrosis factor ligand superfamily member 13B Human genes 0.000 description 1
- 108091005906 Type I transmembrane proteins Proteins 0.000 description 1
- BURPTJBFWIOHEY-UWJYBYFXSA-N Tyr-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 BURPTJBFWIOHEY-UWJYBYFXSA-N 0.000 description 1
- CDRYEAWHKJSGAF-BPNCWPANSA-N Tyr-Ala-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O CDRYEAWHKJSGAF-BPNCWPANSA-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
- NKUGCYDFQKFVOJ-JYJNAYRXSA-N Tyr-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NKUGCYDFQKFVOJ-JYJNAYRXSA-N 0.000 description 1
- 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 1
- JIODCDXKCJRMEH-NHCYSSNCSA-N Val-Arg-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N JIODCDXKCJRMEH-NHCYSSNCSA-N 0.000 description 1
- XGJLNBNZNMVJRS-NRPADANISA-N Val-Glu-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O XGJLNBNZNMVJRS-NRPADANISA-N 0.000 description 1
- 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 1
- DJEVQCWNMQOABE-RCOVLWMOSA-N Val-Gly-Asp Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N DJEVQCWNMQOABE-RCOVLWMOSA-N 0.000 description 1
- DIOSYUIWOQCXNR-ONGXEEELSA-N Val-Lys-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O DIOSYUIWOQCXNR-ONGXEEELSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 230000001745 anti-biotin effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 235000019868 cocoa butter Nutrition 0.000 description 1
- 229940110456 cocoa butter Drugs 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000029578 entry into host Effects 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 239000001761 ethyl methyl cellulose Substances 0.000 description 1
- 235000010944 ethyl methyl cellulose Nutrition 0.000 description 1
- 230000017188 evasion or tolerance of host immune response Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000005182 global health Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108010049041 glutamylalanine Proteins 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
- 239000001963 growth medium Substances 0.000 description 1
- SHFKGANKURXVMY-LCWPZEQJSA-N hcv e2 protein Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)CNC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)O)[C@@H](C)O)C(C)C)[C@@H](C)O)[C@@H](C)O)[C@@H](C)O)CC1=CC=CC=C1 SHFKGANKURXVMY-LCWPZEQJSA-N 0.000 description 1
- 208000005252 hepatitis A Diseases 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 206010019847 hepatosplenomegaly Diseases 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 238000013537 high throughput screening Methods 0.000 description 1
- 108010085325 histidylproline Proteins 0.000 description 1
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 1
- 229950010245 ibalizumab Drugs 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000011866 long-term treatment Methods 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 229960000402 palivizumab Drugs 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 210000001948 pro-b lymphocyte Anatomy 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 108010029020 prolylglycine Proteins 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000029610 recognition of host Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 208000030925 respiratory syncytial virus infectious disease Diseases 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 229960000329 ribavirin Drugs 0.000 description 1
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 1
- 238000002702 ribosome display Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 102000014898 transaminase activity proteins Human genes 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 210000005253 yeast cell Anatomy 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0684—Cells of the urinary tract or kidneys
- C12N5/0686—Kidney cells
-
- 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
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5767—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis non-A, non-B hepatitis
-
- 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
- C07K2317/50—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
- C07K2317/70—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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2510/00—Genetically modified cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2800/00—Nucleic acids vectors
- C12N2800/10—Plasmid DNA
- C12N2800/106—Plasmid DNA for vertebrates
- C12N2800/107—Plasmid DNA for vertebrates for mammalian
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A90/00—Technologies having an indirect contribution to adaptation to climate change
- Y02A90/10—Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Communicable Diseases (AREA)
- Cell Biology (AREA)
- General Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Physics & Mathematics (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Food Science & Technology (AREA)
- Plant Pathology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Oncology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof, belonging to the technical field of biological engineering. The amino acid sequence of the heavy chain variable region of the monoclonal antibody or the fragment thereof is shown as SEQ ID NO. 10; the amino acid sequence of the light chain variable region is shown as SEQ ID NO: 17. The invention also provides a coding gene, an expression vector, application and a composition of the antibody. The binding molecule of the invention can prevent hepatitis C virus from infecting susceptible cells, is fully human, has low immunogenicity and good affinity compared with other animal-derived (such as murine-derived) anti-hepatitis C virus molecules, has good treatment effect and low side effect, and simultaneously provides great guarantee for the standardized production of patent drugs of the antibody.
Description
Technical Field
The invention relates to the technical field of biological engineering, in particular to a fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof.
Background
In the eighties of the last century, researchers first isolated a hepatitis virus, distinct from hepatitis A and B, from the blood of chimpanzees, and this virus was subsequently named Hepatitis C Virus (HCV). Hepatitis c infection has become a global health problem, and people of different genders, ages, and races are susceptible to HCV. The current worldwide HCV infection is estimated to be more than 1.84 million people, and the global HCV infection rate estimated by seropositive rates is about 1.5% -3.5%, while most of the HCV-infected people live in the eastern and southern regions of asia, with about 5,000 million people (total number exceeding 1 million) in both regions. According to the estimation, the prevalence rate of hepatitis C in China is 1%, more than 1000 thousands of HCV carriers exist, and the number of hepatitis C patients is the first in the world.
The prevalence distribution in various regions varies widely around the globe due to the high degree of variation of HCV and immune escape mechanisms. At present, the hepatitis C virus strains which are mainly prevalent all over the world are mainly divided into 11 genotypes and more than 70 subtypes, and HCV viruses with different genotypes have different virulence and great difference in pathogenic capability. Through analysis of results of sample 32030 patients in 29 provinces and cities in China, hepatitis C subtypes which are mainly prevalent in China are 1b (n-16713, 52.18%), 2a (n-9188, 28.69%), 3b (n-2261, 7.06%), 6a (n-2052, 6.41%) and 3a (n-1479, 4.62%), and mixed infection of the main subtypes is found in other small part samples, and the combination is as follows: 1b-2a, 1b-3b, 1b-6a, 3a-3b, 1b-3a and 2a-6a, and the distribution of these HCV subtypes is closely related to the gender, age and geographical location of the population.
According to long-term clinical practice and laboratory technical tests, the clinical diagnosis standard of hepatitis c can be as follows: the clinical manifestations are general hypodynamia, anorexia, right monster costa discomfort, hepatosplenomegaly and so on; the laboratory tests show that glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase are slightly and moderately increased, and the anti-HCV antibody is positive and the HCV RNA is positive; further diagnosing according to the duration of the disease course of the patient and the existence of corresponding epidemiological history.
The main purpose of hepatitis C treatment is to eliminate viruses, keep the replication level of HCV in vivo at an extremely low load for a long time, fully relieve liver injury lesions and the like caused by HCV infection, and improve the long-term survival rate and the survival quality of patients. In the long-term hepatitis C treatment, the clinical classical treatment method is interferon combined with ribavirin, and the method has large side effect, is easy to generate drug resistance and has large effect difference on patients with different genotypes. With the advent of international novel drugs such as DAAs (Direct Anti-viral Agents) such as Dalatavir hydrochloride tablets and Soft Ashbyr capsules, most DAAs have been approved in China for clinical use, the main action mechanism of DAA drugs is to inhibit NS3/4A protease and NS5A/B polymerase in HCV particles, and the HCV infection can be effectively treated by identifying the HCV genotype infected by the patient before the Direct antiviral drugs are used and selecting a proper antiviral scheme in combination with other conditions of liver injury of the patient. In 2018, the 'propanesar' produced by Jilide corporation (Sofosbuweiweipatatavir tablets) formally enters the medicine catalog of China, and marks the coming of a new hepatitis C treatment era, and the new oral hepatitis C treatment medicine with pan-genotype and single tablet brings good news to patients, but the inevitable drug resistance and higher price thereof make some patients go against the battle.
In 1975, Milstein andthe hybridoma technology is invented for the first time, and the technical principle is that immunized mouse spleen cells and myeloma cells are fused, and hybridoma cells capable of stably secreting single antibodies are screened out after fusion. Meanwhile, the concept of the monoclonal antibody is also developed, and the antibody is an antibody which is highly uniform and only aims at a certain specific epitope and is generated by a single B cell clone, and has the advanced advantages of high purity, strong specificity, less cross reaction and the like. However, since the method produces the mouse-derived protein, the method has considerable immunogenicity to human bodies and cannot meet the requirement of long-term treatment. With the development of modern molecular biology and protein engineering techniques, the limitations of the techniques have been addressed to varying degrees. Initially, researchers achieved the production of novel "chimeric" antibodies by substituting a Fragment crystalline Fragment (Fc) sequence of human origin for most of the murine Fc sequence. Later, the degree of humanization of monoclonal antibodies has been improved by further bioengineering murine antigen binding regions (fabs) grafted onto human immunoglobulin (IgG) frameworks, and today mature technology has enabled the production of fully human monoclonal antibodies.
At present, the technology of preparing the monoclonal antibody of the whole human source is various. There are mainly antibody library technologies, such as phage display antibody library technology, ribosome display antibody library technology, single cell cloning expression technology, etc. These techniques are superior and inferior, the antibody library technique can directly prepare specific and stable monoclonal antibodies in vitro, however, the previous library construction workload is very large, the required library capacity needs to cover the antibody diversity of certain animals, high-throughput screening can only obtain partial fragments of the antibodies, and the high affinity of the antibodies cannot be ensured at the later stage. The single cell cloning expression process includes flow cytometry to obtain specific B cell, and gene technology to separate the heavy and light chain variable region gene of the homologous antibody for expression in eukaryotic system. In this method, sorting of pre-B cells is particularly critical and requires highly specific and stable antigens as probes to be effectively sorted.
Monoclonal antibody drugs have been a hot spot for drug development in the last decade. As a novel macromolecular protein medicine, the monoclonal antibody has the advantages of strong specificity and obvious effect, and simultaneously, because the humanization level of the monoclonal antibody medicine is higher, the immunogenicity is relatively lower, the human body is not easy to generate rejection reaction, and the safety is greatly ensured. With the wide application of monoclonal antibodies in the tumor field, the research and development of monoclonal antibody drugs corresponding to infectious diseases are also actively promoted, palivizumab for respiratory syncytial virus infection and ibalizumab for resisting HIV infection are successively produced, and monoclonal antibody drugs for treating hepatitis C are not yet produced.
Disclosure of Invention
The inventor selects E2 protein with good immunogenicity on the HCV surface as an epitope through intensive research, extracts memory B cells from whole blood of rehabilitation people infected with hepatitis C virus pathogens, activates the cells into plasma cells in vitro, screens the plasma cells secreting specific antibodies by adopting a specific antigen, and screens specific antibody genes by utilizing the technologies of efficient extraction of a small amount of cell RNA, nested PCR and the like. The monoclonal antibody is of full human origin, and the variable regions and the constant regions of the heavy chain and the light chain of the monoclonal antibody are all derived from human genes, so the monoclonal antibody has the characteristics of low immunogenicity and high safety.
HCV is a small, enveloped, single-stranded RNA virus with a genome of 9.6kb in length. The HCV particle consists of a positive RNA genome with 5 'and 3' untranslated regions (UTRs) and an Open Reading Frame (ORF) encoding a protein precursor of approximately 3000 amino acids. The UTRs constitute highly conserved cis-acting RNA elements that regulate translation and replication of the viral genome. After assembly processing via the protein precursors encoded by the open reading frame, 10 structural and non-structural proteins were mainly formed (core, E1, E2, p7, NS2, NS3, NS4A, NS4B, NS5A and NS5B proteins). The outer envelope of HCV particle is mainly composed of E1 and E2 glycoprotein, the coding regions of the two proteins are respectively positioned at residue 192-383 site (E1) and 384-746 site (E2) at the N end of the HCV protein structure, and are used as highly glycosylated type I transmembrane proteins which can be combined to form a non-covalent heterodimer structure and have important functions on virus recognition and invasion of host cells. The (core) core protein of HCV particles, a multifunctional protein, has a major role in forming viral capsids, and plays a role in encapsulating and protecting genomic RNA when viruses are transferred between host cells. Other nonstructural proteins such as NS2 and NS3 have been shown to have RNA helicase activity and serine protease activity, and can assist in replication and assembly of viral genome together with other nonstructural proteins. The E1 and E2 proteins, which are envelope proteins on the surface of the virus, are the major viral antigens that elicit a protective immune response. The molecular weight of the two proteins is greatly different, the number of amino acids of the E2 protein is far more than that of the E1 protein, but compared with the E1 protein, the number of hypervariable regions (HVRs) of the E2 protein is less, and the protein structure is more stable, so that when an antigen target is selected, the E2 protein is selected as an epitope, and finally, the fully human monoclonal antibody with High safety and High affinity for the hepatitis C virus is obtained.
In one aspect, the present invention provides a high affinity fully human monoclonal antibody against hepatitis c virus having a heavy chain variable region and a light chain variable region:
(I) the amino acid sequence of the heavy chain variable region is shown as any one of SEQ ID NO 10-13; or has an amino acid sequence with the same function formed by replacing, deleting or adding one or more amino acids in the amino acid sequence shown in any one of SEQ ID NO 10-13;
(II) the light chain variable region has an amino acid sequence as shown in any one of SEQ ID NO 14-18; or an amino acid sequence with the same function formed by replacing, deleting or adding one or more amino acids in the amino acid sequence shown in any one of SEQ ID NO 14-18.
Preferably, the monoclonal antibody has any one of the following groups of heavy chain variable regions and light chain variable regions:
(i) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:10 (5H) 12 ) (ii) a The amino acid sequence of the light chain variable region is shown in SEQ ID NO:14 (7 kappa) 1 );
(ii) The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:10 (5H) 12 ) (ii) a The amino acid sequence of the light chain variable region is shown in SEQ ID NO:17 (7 kappa) 13 );
(iii) The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:11 (5H) 17 ) (ii) a The amino acid sequence of the light chain variable region is shown in SEQ ID NO:15 (7 kappa) 2 );
(iv) The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:12 (5H) 20 ) (ii) a The amino acid sequence of the light chain variable region is shown in SEQ ID NO:18 (7 kappa) 30 );
(v) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 13 (5H) 24 ) (ii) a The amino acid sequence of the light chain variable region is shown as SEQ ID NO:16 (7 kappa) 9 )。
The monoclonal antibody composed of the heavy chain variable region and the light chain variable region can be specifically combined with hepatitis C virus E2 protein.
In another aspect of the present invention, there is provided a nucleic acid molecule encoding the monoclonal antibody described above.
Preferably, the nucleic acid molecule has the nucleotide sequence of the heavy chain variable region as shown in any one of SEQ ID NO 1-4; and the variable region of the light chain as set forth in any one of SEQ ID Nos 5 to 9.
More preferably, the nucleic acid molecule has any one of the following sets of heavy chain variable region and light chain variable region:
(i) the nucleotide sequence of the heavy chain variable region is shown as SEQ ID NO:1 (5H) 12 ) (ii) a The variable region of the light chain has the nucleotide sequence shown in SEQ ID NO:5 (7 kappa) 1 );
(ii) The nucleotide sequence of the heavy chain variable region is shown as SEQ ID NO:1 (5H) 12 ) (ii) a The variable region of the light chain has the nucleotide sequence shown in SEQ ID NO:8 (7 kappa) 13 );
(iii) Of the variable region of the heavy chainThe nucleotide sequence is shown as SEQ ID NO:2 (5H) 17 ) (ii) a The variable region of the light chain has the nucleotide sequence shown in SEQ ID NO:6 (7 kappa) 2 );
(iv) The nucleotide sequence of the heavy chain variable region is shown as SEQ ID NO:3 (5H) 20 ) (ii) a The variable region of the light chain has the nucleotide sequence shown in SEQ ID NO. 9 (7 kappa) 30 );
(v) The nucleotide sequence of the heavy chain variable region is shown as SEQ ID NO. 4 (5H) 24 ) (ii) a The variable region in the light chain has the nucleotide sequence shown in SEQ ID NO:7 (7 kappa) 9 )。
In another aspect of the invention, there is provided an expression vector comprising the nucleic acid molecule described above.
The expression vector may contain, in addition to the nucleic acid molecule described above, a suitable promoter or control sequence. The vector may be used to transform an appropriate host cell so that it can express the protein.
In another aspect of the present invention, there is provided a host cell comprising the above-described expression vector.
The host cell may be a prokaryotic cell, such as a bacterial cell; or lower eukaryotic cells, such as yeast cells; or higher eukaryotic cells, such as mammalian cells. Representative examples are: bacterial cells such as E.coli, Streptomyces; salmonella typhimurium; fungal cells such as yeast; a plant cell; insect cells such as Drosophila S2 or Sf 9; animal cells such as CHO, COS7, NSO or Bowes melanoma cells, etc. Particularly suitable host cells for use in the present invention are eukaryotic host cells, especially mammalian cells, such as 293 cells.
In another aspect of the present invention, there is provided a method for preparing the monoclonal antibody, comprising the steps of:
(1) memory B cell sorting: separating peripheral blood mononuclear cells from a blood sample of a hepatitis C rehabilitation patient, and sorting the peripheral blood mononuclear cells to obtain memory B cells;
(2) in vitro activation culture and positive well screening of memory B cells: activating the memory B cells into plasma cells, detecting IgG antibodies secreted by the memory B cells after the memory B cells are activated into the plasma cells in vitro by ELISA, and screening to obtain positive holes;
(3) constructing and screening an antibody variable region gene library: carrying out reverse transcription on the positive hole cells to obtain cDNA, and amplifying genes of heavy chain variable regions and light chain variable regions of the antibodies;
(4) constructing a library of antibody gene heavy chain variable regions and light chain variable regions: respectively connecting the amplified genes of the heavy chain variable region and the light chain variable region of the antibody to an expression vector;
(5) screening of antibody heavy chain variable region and light chain variable region genes: the recombinant plasmid was co-transfected into HEK293T cells for expression.
In another aspect of the present invention, the monoclonal antibody or the binding fragment thereof is provided for use in the preparation of a medicament for detecting, treating, or preventing hepatitis c virus infection.
In another aspect of the present invention, there is provided a composition comprising an antibody mixture of one or more of said monoclonal antibodies in a therapeutically effective amount, and a pharmaceutically acceptable carrier.
The term "pharmaceutically acceptable" as used herein means that the molecular entities and compositions do not produce adverse, allergic, or other untoward reactions when properly administered to an animal or human. As used herein, a "pharmaceutically acceptable carrier" should be compatible with, i.e., capable of being blended with, the monoclonal antibody of the invention or fragment thereof without substantially reducing the effectiveness of the composition as is often the case.
Specific examples of some substances that may serve as pharmaceutically acceptable carriers or components thereof are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose; powdered gum tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter; polyols such as propylene glycol, glycerol, sorbitol, mannitol and polyethylene glycol; alginic acid; emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; a colorant; a flavoring agent; tabletting agents, stabilizers; an antioxidant; a preservative; pyrogen-free water; isotonic saline solution; and phosphate buffer, and the like.
The compositions of the present invention may be formulated into various dosage forms as desired, and may be administered by a physician in light of such factors as the type, age, weight and general condition of the patient, the mode of administration, and the like, of the dose to be administered to the patient. Administration may be by injection or other therapeutic means, for example.
The pharmaceutical composition may comprise two or more monoclonal antibodies or fragments thereof having neutralizing activity against hepatitis c virus.
In another aspect of the present invention, there is provided a kit for detecting hepatitis c virus, which comprises the monoclonal antibody or a fragment thereof.
Based on the monoclonal antibody or the fragment thereof, a kit for conveniently, quickly and accurately detecting the hepatitis C virus can be prepared. As one detection method of the present invention, an indirect ELISA method is used, in which an antigen to be detected is coated on a solid phase carrier, and detection is performed using the monoclonal antibody or the fragment thereof of the present invention. In a preferred embodiment of the present invention, the monoclonal antibody or the fragment thereof is an antibody, and is detected according to the principle of the double antibody sandwich method. The double antibody sandwich method is conventionally performed by immobilizing a primary antibody (e.g., the monoclonal antibody of the present invention) on a carrier, reacting the primary antibody with an antigen, washing, reacting with a secondary antibody (the secondary antibody carries a detectable signal or can bind to a substance carrying a detectable signal), and detecting a signal by a chemiluminescent or enzyme-linked chromogenic reaction. The double antibody sandwich method is particularly suitable for the detection of antigens having two or more epitopes.
For convenience in detection, the kit may further comprise, in addition to the monoclonal antibody or fragment thereof of the present invention, other detection reagents or auxiliary reagents, such as those conventionally used in ELISA kits, the properties of which and their formulation methods are well known to those skilled in the art, such as color developing agents, labels, secondary antibodies, anti-antibodies, sensitizers, and the like. It will be understood by those skilled in the art that various modifications of the detection kit are encompassed in the present invention, as long as the monoclonal antibody or fragment thereof of the present invention is utilized therein as an agent for recognizing hepatitis C virus.
In addition, instructions for use may be included in the kit to instruct the method of use of the reagents loaded therein.
After obtaining the monoclonal antibody or the fragment thereof provided by the present invention, various immunology-related methods can be used to detect the HCV-E2 protein in the sample, so as to determine whether the donor of the sample to be tested is infected with the hepatitis C virus, and these methods are all included in the present invention. Preferably, the method is for the purpose of non-disease diagnosis.
In another aspect of the invention, there is provided a method of non-therapeutically inhibiting hepatitis c virus, said method comprising administering to a patient an effective amount of said monoclonal antibody or fragment thereof.
In another aspect of the present invention, there is provided a method for non-therapeutic detection of hepatitis c virus, wherein said monoclonal antibody or fragment thereof is contacted with a test sample, and the presence and amount of hepatitis c virus is detected by detecting the binding of said monoclonal antibody or fragment thereof to the test sample.
As used herein, the term "test sample" encompasses a variety of sample types, including blood and other bodily fluid samples of biological origin, solid tissue samples such as biopsy tissue samples or tissue cultures, or cells derived therefrom or progeny thereof. The term also includes samples that have been treated by any means after they have been obtained, for example by treatment with reagents, solubilization, or enrichment for certain components such as proteins or polynucleotides. The term encompasses various clinical samples obtained from any species, also including cultured cells, cell supernatants and cell lysates.
Has the beneficial effects that: compared with the prior art, the fully human monoclonal antibody for resisting the hepatitis C virus is successfully prepared, has the characteristic of high affinity, can be specifically combined with the hepatitis C virus, and can obviously resist the hepatitis C virus. Compared with a mouse antibody, the gene of the fully human antibody is completely derived from human genes, has no components of other species, does not generate toxic or side effects such as anti-mouse antibody and the like in a human body, has better biocompatibility, is more suitable and has more potential to become a macromolecular drug for treating hepatitis C virus, and provides great guarantee for the standardized production of patent drugs of the antibodies.
Drawings
Figure 1ELISA screen positive memory B cell wells.
FIG. 2 is the electrophoresis diagram of the PCR amplification of the heavy chain variable region gene H, H' and the light chain variable region gene kappa, lambda chain of the antibody.
FIG. 3 shows the results of electrophoretic verification of the heavy chain variable region gene, the light chain variable region gene and the vector.
FIG. 4 shows the results of electrophoretic verification of the recombinant plasmid.
FIG. 5 shows a screening scheme for heavy and light chain variable region genes of antibodies.
FIG. 6 shows the results of screening the combination of heavy and light chain variable region genes of antibodies.
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.
Experimental procedures without specific conditions noted in the examples, generally following conventional conditions, such as Sambrook et al, molecular cloning: conditions described in the laboratory Manual (New York: Cold Spring harbor laboratory Press,1989), or according to the manufacturer's recommendations.
Example 1: preparation of fully human monoclonal antibody of hepatitis C virus
1. Sorting of memory B cells
1.1 acquisition of peripheral blood mononuclear cells
Blood is collected by professionals in a sterile environment, and fresh whole blood of hepatitis C recovery volunteers is extracted into disposable vacuum blood collection tubes, wherein the whole blood volume required by each person is 50 ml. The pre-cooled PBS solution after filtration through a 0.22 μm filter was diluted 1:1 with fresh anticoagulated whole blood. The diluted blood was slowly added to an equal volume of lymphocyte separation solution and centrifuged at 2000rpm, Acc4, Dec4 at room temperature for 30 min. After the centrifugation, the centrifuge tube was slowly removed, and the centrifuged second cloudy mononuclear cell layer was carefully aspirated into a 15ml centrifuge tube previously filled with a pre-cooled PBS solution. Centrifuging the sucked mononuclear cells at 1200rpm for 5min at room temperature, discarding the supernatant, and gently blowing and washing with a precooled PBS solution. The resulting solution was centrifuged again, and the collected cells were Peripheral Blood Mononuclear Cells (PBMC) and counted.
1.2 memory B cell sorting
(after cell counting, specific MS or LS columns were selected based on the PBMC obtained from the separation) the cells were centrifuged at 300g for 10min and the supernatant was removed completely. At a rate of 1 × 10 7 The amount of cells was 40. mu.l buffer, and the appropriate amount of buffer was proportionally pipetted to resuspend the cell pellet. At a rate of 1 × 10 7 The cells were added with 10. mu.l of nasal B cell Biotin-Antibody Cocktail as a unit, the cell pellet was mixed well and incubated in a refrigerator at 4 ℃ for 5 min. At a rate of 1 × 10 7 The cell pellet was mixed well with 30. mu.l of buffer and 20. mu.l of Anti-Biotin Micro Beads, and left in a refrigerator at 4 ℃ for 10 min. Adding 1-2 ml buffer solution to wash the cells, centrifuging for 10min at 300g, and discarding the supernatant. The cell pellet was resuspended with 500. mu.l buffer. The column was placed on a magnet and the column was washed with the appropriate volume of buffer (3 ml for LS column and 500. mu.l for MS column). The cell suspension was slowly added to the column and the eluted unlabeled cells were collected. The column was washed 3 times with the appropriate amount of buffer solution (3 ml volume buffer in LS column and 500. mu.l volume buffer in MS column) and the wash solution was added after the column was drained during the wash. Collecting all the eluent to obtain the suspension containing B cells. The column was removed from the separator, placed on a collection tube, and buffer was pipetted into the column and the plunger immediately forced to wash out the magnetic bead labeled cells. Counting cells, centrifuging at 300g for 10min, and completely removing supernatant every 10min 7 Total cells were resuspended in cell pellet with 80. mu.l buffer. To each end1×10 7 Adding a proper amount of labeled magnetic beads into the cells in units of 20 mu lCD27 Microbeads, fully mixing the solution, and incubating for 15min in a refrigerator at 4 ℃. At a rate of 1 × 10 7 The cells were washed in units of cell number plus 1ml buffer, centrifuged at 300g for 10min, and the supernatant was removed completely. When the number of cells is less than 1X 10 8 At this time, the cells were resuspended in 500. mu.l of buffer (if more cells are present, the buffer is doubled). The post was placed on the magnet. The column was washed with the appropriate volume of buffer. The cell suspension was applied to the column. The flow-through unlabeled cells were collected and the column was washed 3 times with the appropriate volume of buffer (3 ml for LS column and 500. mu.l for MS column). Collecting all the eluent, namely the suspension containing the unlabeled cells, and adding the washing liquid after the liquid in the column is drained in the washing process. The column was removed from the separator, immediately placed on a collection tube, an appropriate volume of buffer was pipetted into the column, and the plunger was immediately forced to wash out the magnetic bead labeled cells. The cell is a memory B cell. The sorted memory B cells were counted and cryopreserved.
2. In-vitro activation culture and positive hole screening of memory B cells
And (4) inoculating the sorted memory B cells into a 96-well cell culture plate according to different cell density gradients. Mu.l of 10% FBS IMDM was added to each well as a base medium containing an appropriate concentration of IL-21 as a B cell activating factor. On the basis, a stable cell line of the Co-60 inactivated 3T3-CD40L cells is added into each well to be used as a feeder layer cell. Culturing at 37 deg.C under 5% CO2 for several days to activate memory B cell to become plasma cell and secrete antibody; ELISA detection of secreted IgG antibodies after activation of memory B cells into plasma cells in vitro: cell culture supernatants were collected, coated with a microplate containing Goat Anti-Human Kappa protein and Goat Anti-Human Lambda protein, and the culture supernatants of memory B cells were assayed by ELISA to determine the amount of IgG antibody secreted.
Dissolving Goat Anti-Human Kappa protein and Goat Anti-Human Lambda protein in 100 ng/well of 50 mul/well of coating solution to coat an enzyme label plate respectively, standing overnight at 4 ℃, washing the plate for 5 times by a plate washing machine, wherein the washing solution is PBS solution containing 0.05% Tween-20; sealing with 200 μ l/hole of sealing solution at 37 deg.C for 2h, and washing for 5 times; respectively adding cell culture supernatant or IgG standard substance diluted in gradient into corresponding wells, simultaneously using water or PBS as negative control, standing at 50 μ l/well at 37 deg.C for 1h, and washing for 5 times; adding 1:5000 diluted peroxidase-labeled Goat Anti-Human IgG antibody and 1:10000 diluted Goat x-Human IgG-Fc Fragment HRP Conjugated 50 μ l/well into corresponding wells, standing at 37 deg.C for 45min, and washing for 5 times; adding 100 μ l/hole of color developing solution, and developing at room temperature in dark place for 15 min; adding 2mol/L sulfuric acid 50 mu L/hole, stopping the color reaction, and detecting the OD value of the absorbance of each hole under the condition that the wavelength is 450 nm. And finally, determining positive memory B cell wells by comparing and analyzing the OD value of each well and the corresponding inoculation number of the memory B cells of each well. As can be seen from FIG. 1, in which A1 and A2 are feeder cells as control wells and the OD value is 0.0613, 12 positive wells were finally determined to be numbered A3-A14, wherein the OD values of three positive wells numbered A3, A6 and A7 are significantly higher than those of the other wells, and the OD values are 0.731, 0.4003 and 0.4179, respectively. The rest positive holes are also increased to a certain degree, and the OD values are all about 0.13.
3. Construction and screening of antibody variable region gene library
3.1 Positive well B cell RNA extraction
After microscopic observation, the cell culture supernatant was carefully aspirated away, washed once with PBS, digested with 40. mu.l of 0.25% trypsin per well, and the digestion was observed under a microscope, then the digestion was stopped with 5. mu.l of fetal calf serum per well, the digested cells were blown down and scraped with a pipette tip, and finally 2.25. mu.l of RNase inhibitor (40U/. mu.l) was added per tube. Quickly putting the digested cells into liquid nitrogen for more than 2 min; placing in a PCR instrument at 98 deg.C for 3min, and immediately placing in liquid nitrogen; after the quick freezing, 0.5. mu.l of proteinase K (20mg/ml) and 2.25. mu.l of RNase inhibitor (40U/. mu.l) were added to each tube, and the mixture was placed in a PCR apparatus at 53 ℃ for 1 hour. After the concentration of the extracted RNA was measured, cDNA was obtained by reverse transcription using Promega reverse transcription kit.
3.2 amplification of genes for heavy and light chain variable regions of the antibody
Amplifying antibody heavy chain variable region gene VH by a two-step RT-PCR method (in order to improve the experimental efficiency of each step, heavy chain variable region gene VH primers are divided into two categories of H and H', then PCR amplification and subsequent experiments are respectively carried out, and the two categories are mixed together to pick out single clone when recombinant plasmid is transformed) and light chain variable region gene V kappa or V lambda: taking the cDNA after reverse transcription as a PCR template to carry out a first reaction of nested PCR; then, the first reaction product of the nested PCR is used as a template to perform the second reaction of the nested PCR. Specific primer sequences, PCR reaction procedures, and PCR reaction system references (Nat Protoc. 2009; 4(3): 372-84.).
The PCR products of the heavy chain VH and light chains Vkappa and Vlambda were detected by agarose gel electrophoresis, the results are shown in FIG. 2, FIG. 2A: the results of two rounds of nested PCR amplification of antibody heavy chain (H, H' chain) variable region sequences; b, performing nested PCR two-round amplification on variable region sequences of antibody light chains (kappa chains and lambda chains); c: the results of nested PCR amplification verification of the heavy chain and light chain variable region sequences of samples A3, A4 and A7. M: marker DL 2000. The gel recovery target strip is specifically operated according to the instruction of the OMEGA gel recovery kit. Finally, the concentration is measured by using Nanodrop2000, and the product is stored for a long time at the temperature of-20 ℃. The result shows that the samples A3, A4 and A7 are subjected to two nested PCR amplifications (the kappa chain of the sample A3 is not amplified) to detect obvious target bands, and the positions of the target bands are between 250bp and 500bp, so that the samples A3, A4 and A7 are determined to be selected for subsequent experiments.
3.3 construction of library of heavy chain variable regions and light chain variable regions of antibody Gene
The amplified heavy chain variable region gene VH and light chain variable region genes Vkappa and Vlambda were ligated to expression vectors pIgH (AbVec-hIgG1 for VH), pIgK (AbVec-hIgKappa for Vkappa) and pIg lambda (IG-lambda expression vector for V lambda), respectively (NCBI GenBank accession Nos.: FJ475055, FJ475056, FJ 517647).
Sequentially using AgeI-HF and Sal I-HF to carry out enzyme digestion on VH and pIgH; cutting V lambda and pIg lambda by AgeI-HF and Xho I; the V kappa and the pIg kappa are cut by AgeI-HF, and then the V kappa and the pIg kappa are cut by BsiW I. The enzyme digestion product is detected by 1.2 percent agarose gel electrophoresis, the band is observed, the target gene band is at the position of 300bp-500bp, and the experimental result is shown in figure 3. The gel recovery target strip is specifically operated according to the instruction of the OMEGA gel recovery kit.
FIG. 3 shows the results of electrophoretic verification of the heavy chain variable region gene, the light chain variable region gene and the vector. Wherein VH, VH', V lambda and V kappa are all enzyme digestion verification. A: the enzyme digestion results of the variable region genes of sample heavy chains (H chain, H' chain) A3, A4 and A7; b: the results of the enzyme digestion of the variable region genes of No. A3, A4 and A7 light chain (kappa chain and lambda chain); c: cleavage of the vector plasmid (IgH, Ig κ, Ig λ), M: marker DL2000, M 2 :Marker DL10000。
According to the size of the target fragment, namely the gene fragment of the heavy chain and light chain variable regions of the antibody is 400bp, and the corresponding plasmid of the heavy chain and light chain vector is 5000bp, the successful enzyme digestion of the heavy chain and light chain variable regions of the A3, A4 and A7-like antibodies and the plasmid of the vector can be judged.
The target gene fragment after digestion is ligated with the corresponding constant region vector pIgH (AbVec-hIgG1 for VH), pIgkK (AbVec-hIgKappa for V kappa) or pIg lambda (IG-lambda expression vector for V lambda) after digestion, and enzymatically ligated at 16 ℃ overnight.
The ligated recombinant plasmid was transformed into TOP10 E.coli cells. The next day, positive clone colonies were picked up in Amp-containing cells + After shaking culture at 37 ℃ for 12h at 200rpm in LB medium, 50% sterilized glycerol was used for conservation of bacteria, and plasmids were extracted (see the OMEGA plasmid extraction kit for details). And (3) enzyme digestion verification of the positive clonobacterium antibody vector plasmids of the heavy chain H-IgH, the H' -IgH and the light chain lambda-Ig lambda. The experimental results are shown in FIG. 4, after the recombinant plasmids A3, A4 and A7 are digested, two bands can be observed at the positions of the band above 2000bp (5000bp) and the band 400bp, namely two bands of the plasmid gene and the antibody heavy-light chain variable region gene formed after the recombinant plasmids are digested, which shows that the early-stage A3, A4 and A7 sample heavy-chain plasmid is successfully enzymatically linked (M: DL 2000).
4. Screening of antibody heavy chain variable region and light chain variable region genes
After the recombinant plasmids are transformed, monoclonal plasmids are picked, a certain plasmid of an antibody heavy chain and a certain plasmid of an antibody lambda light chain or an antibody kappa light chain (two light chains are lambda and kappa; only one heavy chain is H, and any one of H, lambda or kappa can be combined to form an antibody) are cotransfected to HEK293T cells, the secretion amount of specific antibodies of HCV-E2 antigen protein (self-made) is detected by an enzyme-linked immunosorbent assay method (Elisa method), and an antibody recombinant plasmid combination with high expression of HCV-E2 specific IgG antibodies is screened.
The specific steps of cell transfection are as follows:
(1) inoculating cells: 0.25% of the cells after termination of the digestion with pancreatin-EDTA were added at 1X 10/well 4 Each cell was inoculated into a 96-well plate and supplemented with culture medium to 190. mu.L. Setting a control group, wherein the control group 1 is added with a transfection reagent and cells but not plasmids; control 2 was cell only, without transfection reagent and plasmid.
(2) The plasmid and transfection reagent were diluted separately with opti-MEM such that 0.15. mu.L of transfection reagent was added per well and 100ng of plasmid was added per well.
(3) According to the following steps: 1, mixing the diluted plasmid and diluted transfection reagent, adding them to the wells inoculated with cells after 5min, 37 deg.C, 5% CO 2 Culturing in an incubator for 24-72h, and detecting the content of the antibody.
(4) Elisa assay cell culture supernatant IgG secretion: collecting the cell culture supernatant after transfection in the step (3), and coating an enzyme label plate with HCV-E2 antigen protein (self-made) according to the coating amount of 100 ng/hole by using a coating solution (50 mu l/hole); the microplate was additionally coated with Goat Anti-Human Kappa + Goat Anti-Human Lambda (100 ng/well each) protein and incubated overnight at 4 ℃. The plate washing machine washes the plate 5 times. The wash solution was a PBS solution containing 0.05% Tween. Blocking with 200. mu.l/well of blocking solution at 37 ℃ for 2h, and washing the plate 5 times. Cell culture supernatants, IgG standards and anti-HCV-E2 protein antibodies (primary antibodies) were added to the corresponding wells, respectively, and the wells were incubated at 37 ℃ for 1 hour with water or PBS solution as a negative control for 5 times at 50. mu.l/well. Add 1:5000 dilution of Goat Anti-Human IgG and Goat Anti-Mouse IgG and 1:10000 dilution of Goat x-Human IgG-Fc Fragment HRP Conjugated 50. mu.l/well to the corresponding wells, leave them at 37 ℃ for 45min, wash the plate 5 times. Adding TMB color developing agent 100 μ l/hole, and developing at room temperature in dark for 20 min. Adding 50 mu L/hole of 2mol/L sulfuric acid, stopping the color reaction, and detecting OD/450nm with the reference wavelength of 630 nm.
Using the screening format shown in FIG. 5, we performed ELISAPositive memory B cell wells were screened for specificity. Ranking is carried out in different batches of screening by taking the antibody affinity as a standard, the optimal heavy-light chain sequence is selected for carrying out next batch of screening after ranking, the monoclonal antibody with high affinity is obtained by screening layer by layer from the mixed sequence to the monoclonal sequence, and the effective monoclonal heavy chain and light chain combination is selected for carrying out the detection of the monoclonal antibody after identification and analysis (sequencing is completed by Jiangsu Kisry Biotech company). Screening to obtain heavy and light chain monoclonal sequences, selecting 5 heavy and light chain monoclonal sequences respectively, and performing in-vitro transfection expression in a one-to-one matching way, wherein the heavy chain monoclonal sequences of the 5 heavy chains: 5H 12 ,5H 16 ,5H 17 ,5H 20 ,5H 24 And 5 light chain monoclonal sequences: 7 kappa 1 ,7κ 2 ,7κ 9 ,7κ 13 ,7κ 30 And finally obtaining 5 monoclonal antibodies with high affinity after ELISA screening, wherein the antibody combination is as follows: 5H 12 +7κ 1 ,5H 12 +7κ 13 ,5H 17 +7κ 2 ,5H 20 +7κ 30 ,5H 24+ 7κ 9 (the results correspond to FIG. 6).
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
SEQUENCE LISTING
<110> university of Nanchang
<120> a fully human monoclonal antibody with high affinity against hepatitis C virus and use thereof
<130> 1
<160> 18
<170> PatentIn version 3.5
<210> 1
<211> 356
<212> DNA
<213> 5H12 heavy chain variable region nucleotide sequence
<400> 1
gtacattccg aggtgcagct ggtgcagtcg gggggaacct tggtacagcc tggggggtcc 60
ctgagactct cttgtgaagc ctctggattc acctttagcg actatgccat gggctgggtc 120
cgccagactc caggaaaggg gctggagtgg ctgtcggcta ttcgtaaaag tggcactacc 180
acatactacg cggactccgt gaagggccgg ttcatcatct ccagagacaa ttccaagaac 240
accctgtatc tgcaaatgaa taggctgagg gtcggcgaca cggccactta ttactgtgcg 300
actcacccca tcgcgggcta ctggggccag ggaaccacgg tcaccgtctc ctcagc 356
<210> 2
<211> 356
<212> DNA
<213> 5H17 heavy chain variable region nucleotide sequence
<400> 2
gtacattccc aggtccagct ggtacagtcg gggggaacct tggtacagcc tggggggtcc 60
ctgagactct cttgtgaagc ctctggattc acctttagca actatgccat gggctgggtc 120
cgccagactc caggaaaggg gctggagtgg ctgtcggcta ttcgtaaaag tggcactacc 180
acatactacg cggactccgt gaagggccgg ttcatcatct ccagagacaa ttccaagaac 240
accctgtatc tgcaaatgaa taggctgagg gtcggcgaca cggccactta ttactgtgcg 300
actcacccca tcgcgggcta ctggggccag ggaaccacgg tcaccgtctc ctcagc 356
<210> 3
<211> 374
<212> DNA
<213> 5H20 heavy chain variable region nucleotide sequence
<400> 3
gtacattcct cccaggtcca gctggtacag tcggggggaa ccttggtaca gcctgggggg 60
tccctgagac tctcttgtga agcctctgga ttcaccttta gcaactatgc catgggctgg 120
gtccgccaga ctccaggaaa ggggctggag tggctgtcgg ctattcgttc caaaagtggc 180
actaccacat actacgcgga ctccgtgaag ggccggttca tcatctccag agacaattcc 240
aagaacaccc tgtatctgca aatgaatagg ctgagggtcg gcgacacggc ctcggggact 300
tattactgtg cgactcaccc catcgcgtcg gggggctact ggggccaggg aaccacggtc 360
accgtctcct cagc 374
<210> 4
<211> 380
<212> DNA
<213> 5H24 heavy chain variable region nucleotide sequence
<400> 4
gtacattccg aggtgcagct ggtgcagtcg gcttcggggg gattgacctt ggtacagcct 60
ggggggtccc tgagactctc ttgtgaagcc tctggattca cctttagcaa ctatgccatg 120
ggctcggctt gggtccgcca gactccagga aaggggctgg agtggctgtc ggcttcggct 180
attcgtaaaa gtggcactac cacatactac gcggactccg tgaagggccg gttcatcatc 240
tccagagaca attccaagaa caccctgtat ctgcaaatga ataggctgag ggtcggcgac 300
acggccactt attactgtgc gactcacccc atcgcgggct actggggcca gggaaccacg 360
gtcaccgtct ttgcctcagc 380
<210> 5
<211> 357
<212> DNA
<213> 7 kappa 1 light chain variable region nucleotide sequence
<400> 5
gtacattcag aaatagtgat gacgcagtct ccagccaccc tgtctgtgtc tagggaccca 60
ggggaaagag ccaccctctc cttagggtgc aggttagggg ccagtcagag tgttagcagc 120
aacttagcct ggtaccggca gaaacctggc caggctccca ggctcctcat ctatggtgca 180
tccaccaggg ccactggtat cccagccagg ttcagtggca gtgggtctgg gacagagttc 240
actctcacca tcagcagcct gcagtctgaa gattttgcag tttattactg tcagcagtat 300
aataactggc ctccgtggac gttagggacc ggccaaggga ccaaggtgga aatcaaa 357
<210> 6
<211> 345
<212> DNA
<213> 7 kappa 2 light chain variable region nucleotide sequence
<400> 6
gtacattcag aaatagtgat gacgcagtct ccagccaccc tgtctgtgtc tccaggggaa 60
agagccaccc tctcctgcag ggccagtcag agtgttagca gcaacttagc ctggtaccgg 120
cagaaacctg gccaggctcc caggctcctc atctatggtg catccaccag ggccactatc 180
tatggtgcag gtatcccagc caggttcagt ggcagtgggt ctgggacaga gttcactctc 240
accatcagca gcctgcagtc tgaagatttt gcagtttatt actgtcagca gtataataac 300
tggcctccgt ggacgttcgg ccaagggacc aaggtggaaa tcaaa 345
<210> 7
<211> 339
<212> DNA
<213> 7 kappa 9 light chain variable region nucleotide sequence
<400> 7
gtacattcag aaatagtgat gacgcagtct ccactgtctg tgtctccagg ggaaagagcc 60
accctctcct gcagggccag tagggccact cagagtgtta gcagcaactt agcctggtac 120
cggcagaaac ctggccaggc tcccaggctc ctcatctatg gtgcatccac cagggccact 180
ccagccaggt tcagtggcag tgggtctggg acagagttca ctctcaccat cagcagcctg 240
cagtctgaag attttgcagt ttattactgt cagcagtata ataactggcc tccgtggacg 300
ttcggccaag ggaccaaggt gagggccact gaaatcaaa 339
<210> 8
<211> 345
<212> DNA
<213> 7 kappa 13 light chain variable region nucleotide sequence
<400> 8
gtacattcag aaatagtgat gacgcagtct tctccaccag ccaccctgtc tgtgtctcca 60
ggggaaagag ccaccctctc ctgcagggcc agtcagagtg ttagcagcaa cttagcctgg 120
taccggcaga aacctggcca ggctcccagg ctcctcatct atggtgcatc caccagggcc 180
actggtatcc cagccaggtt cagtggcagt gggtctggga cagagttcac tctcaccatc 240
agcagcctgc agtctgaaga ttttgcagtt tattactgtc agcagtataa taactggcct 300
ccgtggacgt tcggccaagg gaccaaggtg tctccagaaa tcaaa 345
<210> 9
<211> 363
<212> DNA
<213> 7 kappa 30 light chain variable region nucleotide sequence
<400> 9
gtacattcag aaatagtgat gacgcagtct ccagccccac tggtaaccct gtctgtgtct 60
ccaggggaaa gagccaccct ctcctgcagg gccagtcaga gtgccagtgt tagcagcaac 120
ttagcctggt accggcagaa acctggccag gctcccaggc tcctcatcta tggtgcatcc 180
accagggcca ctggtatccc accactggta gccaggttca gtggcagtgg gtctgggaca 240
gagttcactc tcaccatcag cagcctgcag tctgaagatt ttgcagttta ttactgtcag 300
cagtataata actggcctcc gtggacgttc ggccaaggga ccaaggtgga agccagtatc 360
aaa 363
<210> 10
<211> 115
<212> PRT
<213> 5H12 heavy chain variable region amino acid sequence
<400> 10
Val His Ser Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val
1 5 10 15
Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ala
20 25 30
Ser Val Ser Ser Asn Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala
35 40 45
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro
50 55 60
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile
65 70 75 80
Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
85 90 95
Asn Asn Trp Pro Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ala
100 105 110
Ser Ile Lys
115
<210> 11
<211> 118
<212> PRT
<213> 5H17 heavy chain variable region amino acid sequence
<400> 11
Val His Ser Gln Val Gln Leu Val Gln Ser Gly Gly Thr Leu Val Gln
1 5 10 15
Pro Gly Gly Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe
20 25 30
Ser Asn Tyr Ala Met Gly Trp Val Arg Gln Thr Pro Gly Lys Gly Leu
35 40 45
Glu Trp Leu Ser Ala Ile Arg Lys Ser Gly Thr Thr Thr Tyr Tyr Ala
50 55 60
Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asn
65 70 75 80
Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Val Gly Asp Thr Ala Thr
85 90 95
Tyr Tyr Cys Ala Thr His Pro Ile Ala Gly Tyr Trp Gly Gln Gly Thr
100 105 110
Thr Val Thr Val Ser Ser
115
<210> 12
<211> 124
<212> PRT
<213> 5H20 heavy chain variable region amino acid sequence
<400> 12
Val His Ser Ser Gln Val Gln Leu Val Gln Ser Gly Gly Thr Leu Val
1 5 10 15
Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr
20 25 30
Phe Ser Asn Tyr Ala Met Gly Trp Val Arg Gln Thr Pro Gly Lys Gly
35 40 45
Leu Glu Trp Leu Ser Ala Ile Arg Ser Lys Ser Gly Thr Thr Thr Tyr
50 55 60
Tyr Ala Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser
65 70 75 80
Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Val Gly Asp Thr
85 90 95
Ala Ser Gly Thr Tyr Tyr Cys Ala Thr His Pro Ile Ala Ser Gly Gly
100 105 110
Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 13
<211> 126
<212> PRT
<213> 5H24 heavy chain variable region amino acid sequence
<400> 13
Val His Ser Glu Val Gln Leu Val Gln Ser Ala Ser Gly Gly Leu Thr
1 5 10 15
Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly
20 25 30
Phe Thr Phe Ser Asn Tyr Ala Met Gly Ser Ala Trp Val Arg Gln Thr
35 40 45
Pro Gly Lys Gly Leu Glu Trp Leu Ser Ala Ser Ala Ile Arg Lys Ser
50 55 60
Gly Thr Thr Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Ile Ile
65 70 75 80
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu
85 90 95
Arg Val Gly Asp Thr Ala Thr Tyr Tyr Cys Ala Thr His Pro Ile Ala
100 105 110
Gly Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Phe Ala Ser
115 120 125
<210> 14
<211> 119
<212> PRT
<213> 7 kappa 1 light chain variable region amino acid sequence
<400> 14
Val His Ser Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val
1 5 10 15
Ser Arg Asp Pro Gly Glu Arg Ala Thr Leu Ser Leu Gly Cys Arg Leu
20 25 30
Gly Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Arg Gln Lys
35 40 45
Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala
50 55 60
Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe
65 70 75 80
Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr
85 90 95
Cys Gln Gln Tyr Asn Asn Trp Pro Pro Trp Thr Leu Gly Thr Gly Gln
100 105 110
Gly Thr Lys Val Glu Ile Lys
115
<210> 15
<211> 115
<212> PRT
<213> 7 kappa 2 light chain variable region amino acid sequence
<400> 15
Val His Ser Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val
1 5 10 15
Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val
20 25 30
Ser Ser Asn Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg
35 40 45
Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Ile Tyr Gly Ala Gly
50 55 60
Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu
65 70 75 80
Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln
85 90 95
Gln Tyr Asn Asn Trp Pro Pro Trp Thr Phe Gly Gln Gly Thr Lys Val
100 105 110
Glu Ile Lys
115
<210> 16
<211> 113
<212> PRT
<213> 7 kappa 9 light chain variable region amino acid sequence
<400> 16
Val His Ser Glu Ile Val Met Thr Gln Ser Pro Leu Ser Val Ser Pro
1 5 10 15
Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Arg Ala Thr Gln Ser
20 25 30
Val Ser Ser Asn Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro
35 40 45
Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu
65 70 75 80
Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp
85 90 95
Pro Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Arg Ala Thr Glu Ile
100 105 110
Lys
<210> 17
<211> 115
<212> PRT
<213> 7 kappa 13 light chain variable region amino acid sequence
<400> 17
Val His Ser Glu Ile Val Met Thr Gln Ser Ser Pro Pro Ala Thr Leu
1 5 10 15
Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln
20 25 30
Ser Val Ser Ser Asn Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala
35 40 45
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro
50 55 60
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile
65 70 75 80
Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
85 90 95
Asn Asn Trp Pro Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Ser Pro
100 105 110
Glu Ile Lys
115
<210> 18
<211> 121
<212> PRT
<213> 7 kappa 30 light chain variable region amino acid sequence
<400> 18
Val His Ser Glu Ile Val Met Thr Gln Ser Pro Ala Pro Leu Val Thr
1 5 10 15
Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser
20 25 30
Gln Ser Ala Ser Val Ser Ser Asn Leu Ala Trp Tyr Arg Gln Lys Pro
35 40 45
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr
50 55 60
Gly Ile Pro Pro Leu Val Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr
65 70 75 80
Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val
85 90 95
Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Pro Trp Thr Phe Gly Gln
100 105 110
Gly Thr Lys Val Glu Ala Ser Ile Lys
115 120
Claims (7)
1. A high affinity fully human monoclonal antibody against hepatitis C virus having the following heavy and light chain variable regions:
the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 10; the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 17.
2. The fully human monoclonal antibody of claim 1, wherein the antibody specifically binds hepatitis C virus E2 protein.
3. A nucleic acid molecule encoding the monoclonal antibody of claim 1, having the following heavy chain variable region and light chain variable region:
the nucleotide sequence of the heavy chain variable region is shown as SEQ ID NO: 1; the variable region in the light chain has the nucleotide sequence shown in SEQ ID NO 8.
4. An expression vector comprising the nucleic acid molecule of claim 3.
5. A host cell comprising the expression vector of claim 4.
6. Use of the monoclonal antibody or binding fragment thereof of claim 1 or 2 in the preparation of a medicament for detecting, treating, or preventing hepatitis c virus infection.
7. A composition comprising a therapeutically effective amount of the monoclonal antibody of claim 1, and a pharmaceutically acceptable carrier.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210682852.8A CN115028712B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110498647.1A CN113234147B (en) | 2021-05-08 | 2021-05-08 | Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof |
CN202210682852.8A CN115028712B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110498647.1A Division CN113234147B (en) | 2021-05-08 | 2021-05-08 | Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115028712A true CN115028712A (en) | 2022-09-09 |
CN115028712B CN115028712B (en) | 2024-02-09 |
Family
ID=77132506
Family Applications (5)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110498647.1A Active CN113234147B (en) | 2021-05-08 | 2021-05-08 | Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof |
CN202210682852.8A Active CN115028712B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
CN202210669038.2A Active CN115124617B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
CN202210676544.4A Active CN115028711B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
CN202210661696.7A Active CN114920836B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110498647.1A Active CN113234147B (en) | 2021-05-08 | 2021-05-08 | Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof |
Family Applications After (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210669038.2A Active CN115124617B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
CN202210676544.4A Active CN115028711B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
CN202210661696.7A Active CN114920836B (en) | 2021-05-08 | 2021-05-08 | High-affinity anti-hepatitis C virus fully-humanized monoclonal antibody and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (5) | CN113234147B (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111925434B (en) * | 2020-06-22 | 2023-06-27 | 南昌大学 | Screening method of monoclonal antibody |
CN114195887B (en) * | 2021-11-23 | 2024-02-20 | 武汉瑞法医疗器械有限公司 | Hepatitis B virus surface antigen adsorbent and application thereof |
CN114853886B (en) * | 2022-05-23 | 2023-11-28 | 南昌大学 | Fully human monoclonal antibody with high affinity against human cytomegalovirus and application thereof |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104974246A (en) * | 2014-04-01 | 2015-10-14 | 中国科学院上海巴斯德研究所 | Total-human monoclonal antibody for neutralizing hepatitis C virus and application of total-human monoclonal antibody |
CN105001325A (en) * | 2015-07-31 | 2015-10-28 | 北京泰诺迪生物科技有限公司 | Total-humanized anti-hepatitis B virus neutralizing antibody and preparing method and application thereof |
CN106749644A (en) * | 2016-11-14 | 2017-05-31 | 广州泰诺迪生物科技有限公司 | A kind of neutralizing antibody TRN1001 of full people source HCV-Ab IgG |
CN107011435A (en) * | 2016-06-01 | 2017-08-04 | 苏州银河生物医药有限公司 | The human monoclonal antibodies 8D6 of anti-hepatitis c virus |
CN107286237A (en) * | 2017-08-07 | 2017-10-24 | 广州泰诺迪生物科技有限公司 | The acquisition and application of a kind of anti-HCV |
CN109021098A (en) * | 2018-08-06 | 2018-12-18 | 南京鼓楼医院 | Full Humanized monoclonal antibodies and its preparation method and application |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6747136B2 (en) * | 1996-04-19 | 2004-06-08 | Karolinska Innovations Ab | Human monoclonal antibodies specific for hepatitis C virus (HCV) E2 antigen |
CN111620944B (en) * | 2020-06-01 | 2023-03-21 | 长春师范大学 | Fully humanized anti-hepatitis B virus monoclonal antibody, preparation method and application thereof |
-
2021
- 2021-05-08 CN CN202110498647.1A patent/CN113234147B/en active Active
- 2021-05-08 CN CN202210682852.8A patent/CN115028712B/en active Active
- 2021-05-08 CN CN202210669038.2A patent/CN115124617B/en active Active
- 2021-05-08 CN CN202210676544.4A patent/CN115028711B/en active Active
- 2021-05-08 CN CN202210661696.7A patent/CN114920836B/en active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104974246A (en) * | 2014-04-01 | 2015-10-14 | 中国科学院上海巴斯德研究所 | Total-human monoclonal antibody for neutralizing hepatitis C virus and application of total-human monoclonal antibody |
CN105001325A (en) * | 2015-07-31 | 2015-10-28 | 北京泰诺迪生物科技有限公司 | Total-humanized anti-hepatitis B virus neutralizing antibody and preparing method and application thereof |
CN107011435A (en) * | 2016-06-01 | 2017-08-04 | 苏州银河生物医药有限公司 | The human monoclonal antibodies 8D6 of anti-hepatitis c virus |
CN106749644A (en) * | 2016-11-14 | 2017-05-31 | 广州泰诺迪生物科技有限公司 | A kind of neutralizing antibody TRN1001 of full people source HCV-Ab IgG |
CN107286237A (en) * | 2017-08-07 | 2017-10-24 | 广州泰诺迪生物科技有限公司 | The acquisition and application of a kind of anti-HCV |
CN109021098A (en) * | 2018-08-06 | 2018-12-18 | 南京鼓楼医院 | Full Humanized monoclonal antibodies and its preparation method and application |
Also Published As
Publication number | Publication date |
---|---|
CN114920836A (en) | 2022-08-19 |
CN113234147B (en) | 2022-08-09 |
CN115028712B (en) | 2024-02-09 |
CN113234147A (en) | 2021-08-10 |
CN115028711B (en) | 2024-02-09 |
CN114920836B (en) | 2024-03-29 |
CN115028711A (en) | 2022-09-09 |
CN115124617B (en) | 2024-02-09 |
CN115124617A (en) | 2022-09-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113234147B (en) | Fully human monoclonal antibody with high affinity for resisting hepatitis C virus and application thereof | |
US20090104188A1 (en) | Anti-hepatitis c virus antibody and uses thereof | |
CN115260307B (en) | High-affinity anti-rabies virus fully human monoclonal antibody and application thereof | |
CN109265542B (en) | Antibody specifically binding norovirus GII.4 genotype VP1 protein or VLP, and preparation method and application thereof | |
WO2013043125A1 (en) | Enterovirus 71 specific antibodies and uses thereof | |
Petit et al. | Enveloped particles in the serum of chronic hepatitis C patients | |
ES2377968T3 (en) | Antibodies directed against the hepatitis C virus E1E2 complex and pharmaceutical compositions | |
CN117964746A (en) | Neutralizing antibody HY2 of A-type foot-and-mouth disease virus, preparation method and application thereof | |
CN114316040B (en) | Fully human monoclonal antibody for resisting novel coronavirus and application thereof | |
CN109535249B (en) | Monoclonal antibody ZKns3G2 and application thereof | |
CN109503712B (en) | Monoclonal antibody ZKns2E11 and application thereof | |
US10888615B2 (en) | Neutralizing human monoclonal antibody 8D6 against HCV infection | |
CN104119436B (en) | The neutrality human monoclonal antibodies of anti-hepatitis c virus | |
CN108794625A (en) | A kind of monoclonal antibody of anti-EV-D68 viruses and its preparation and application | |
CN114853886B (en) | Fully human monoclonal antibody with high affinity against human cytomegalovirus and application thereof | |
CN114591427B (en) | Mouse anti-MPT 32 protein hybridoma cell line 13B12, monoclonal antibody based on same and application thereof | |
CN109678955B (en) | Monoclonal antibody pair for detecting HCV-cAg, hybridoma cell strain secreting antibody pair and application of monoclonal antibody pair | |
CN118388642A (en) | Nanometer antibody targeting BVDV structural protein E2 and preparation and application thereof | |
CN115873122A (en) | Light chain and heavy chain variable regions of carbaryl-resistant monoclonal antibody and application thereof |
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 |