KR102259974B1 - Method for producing target antigen-specific antibody using recombinant antigen - Google Patents
Method for producing target antigen-specific antibody using recombinant antigen Download PDFInfo
- Publication number
- KR102259974B1 KR102259974B1 KR1020180137394A KR20180137394A KR102259974B1 KR 102259974 B1 KR102259974 B1 KR 102259974B1 KR 1020180137394 A KR1020180137394 A KR 1020180137394A KR 20180137394 A KR20180137394 A KR 20180137394A KR 102259974 B1 KR102259974 B1 KR 102259974B1
- Authority
- KR
- South Korea
- Prior art keywords
- antigen
- glu
- leu
- lys
- ser
- Prior art date
Links
- 102000036639 antigens Human genes 0.000 title claims abstract description 122
- 108091007433 antigens Proteins 0.000 title claims abstract description 118
- 239000000427 antigen Substances 0.000 title claims abstract description 117
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 40
- 230000004927 fusion Effects 0.000 claims abstract description 77
- 238000000034 method Methods 0.000 claims abstract description 31
- 241000700605 Viruses Species 0.000 claims abstract description 23
- 241000124008 Mammalia Species 0.000 claims abstract description 13
- 239000000203 mixture Substances 0.000 claims abstract description 7
- 208000036142 Viral infection Diseases 0.000 claims abstract description 5
- 230000009385 viral infection Effects 0.000 claims abstract description 5
- 241000699670 Mus sp. Species 0.000 claims description 22
- 241001465754 Metazoa Species 0.000 claims description 14
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 14
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 11
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims description 11
- 230000016784 immunoglobulin production Effects 0.000 claims description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 6
- 102000052866 Amino Acyl-tRNA Synthetases Human genes 0.000 claims description 5
- 108700028939 Amino Acyl-tRNA Synthetases Proteins 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 230000002708 enhancing effect Effects 0.000 claims description 4
- 230000004570 RNA-binding Effects 0.000 claims description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 2
- 241000272517 Anseriformes Species 0.000 claims description 2
- 241000282472 Canis lupus familiaris Species 0.000 claims description 2
- 241000283086 Equidae Species 0.000 claims description 2
- 241000282326 Felis catus Species 0.000 claims description 2
- 241000287828 Gallus gallus Species 0.000 claims description 2
- 235000013330 chicken meat Nutrition 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 230000027455 binding Effects 0.000 abstract description 8
- 238000001514 detection method Methods 0.000 abstract description 8
- 230000001965 increasing effect Effects 0.000 abstract description 8
- 238000011160 research Methods 0.000 abstract description 5
- 230000001225 therapeutic effect Effects 0.000 abstract description 3
- 108090000623 proteins and genes Proteins 0.000 description 80
- 102000004169 proteins and genes Human genes 0.000 description 77
- 235000018102 proteins Nutrition 0.000 description 63
- 238000002965 ELISA Methods 0.000 description 48
- 101710154606 Hemagglutinin Proteins 0.000 description 34
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 34
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 34
- 101710176177 Protein A56 Proteins 0.000 description 34
- 208000025370 Middle East respiratory syndrome Diseases 0.000 description 30
- 238000001262 western blot Methods 0.000 description 21
- 150000001413 amino acids Chemical group 0.000 description 17
- 210000004408 hybridoma Anatomy 0.000 description 17
- 102000037865 fusion proteins Human genes 0.000 description 16
- 108020001507 fusion proteins Proteins 0.000 description 16
- 239000000185 hemagglutinin Substances 0.000 description 16
- 238000011081 inoculation Methods 0.000 description 16
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 description 15
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 15
- 241000699666 Mus <mouse, genus> Species 0.000 description 14
- 241000712461 unidentified influenza virus Species 0.000 description 14
- 210000004027 cell Anatomy 0.000 description 13
- 241000588724 Escherichia coli Species 0.000 description 12
- 108091005634 SARS-CoV-2 receptor-binding domains Proteins 0.000 description 12
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 11
- 238000010586 diagram Methods 0.000 description 10
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 102000017737 Lysine-tRNA Ligase Human genes 0.000 description 9
- 108010092041 Lysine-tRNA Ligase Proteins 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 8
- 108010076818 TEV protease Proteins 0.000 description 7
- 108010005233 alanylglutamic acid Proteins 0.000 description 7
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 7
- 208000037797 influenza A Diseases 0.000 description 7
- 108010009298 lysylglutamic acid Proteins 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 239000013598 vector Substances 0.000 description 7
- JJKKWYQVHRUSDG-GUBZILKMSA-N Glu-Ala-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O JJKKWYQVHRUSDG-GUBZILKMSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 101000908391 Homo sapiens Dipeptidyl peptidase 4 Proteins 0.000 description 6
- 241000880493 Leptailurus serval Species 0.000 description 6
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 6
- 239000002671 adjuvant Substances 0.000 description 6
- 108010028295 histidylhistidine Proteins 0.000 description 6
- 102000045598 human DPP4 Human genes 0.000 description 6
- 108010034529 leucyl-lysine Proteins 0.000 description 6
- 238000007500 overflow downdraw method Methods 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- ZRHDPZAAWLXXIR-SRVKXCTJSA-N Leu-Lys-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O ZRHDPZAAWLXXIR-SRVKXCTJSA-N 0.000 description 5
- JIHDFWWRYHSAQB-GUBZILKMSA-N Leu-Ser-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O JIHDFWWRYHSAQB-GUBZILKMSA-N 0.000 description 5
- RMLLCGYYVZKKRT-CIUDSAMLSA-N Met-Ser-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O RMLLCGYYVZKKRT-CIUDSAMLSA-N 0.000 description 5
- 108010047495 alanylglycine Proteins 0.000 description 5
- 108010047857 aspartylglycine Proteins 0.000 description 5
- 206010022000 influenza Diseases 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 210000004988 splenocyte Anatomy 0.000 description 5
- 108010073969 valyllysine Proteins 0.000 description 5
- OZRFYUJEXYKQDV-UHFFFAOYSA-N 2-[[2-[[2-[(2-amino-3-carboxypropanoyl)amino]-3-carboxypropanoyl]amino]-3-carboxypropanoyl]amino]butanedioic acid Chemical compound OC(=O)CC(N)C(=O)NC(CC(O)=O)C(=O)NC(CC(O)=O)C(=O)NC(CC(O)=O)C(O)=O OZRFYUJEXYKQDV-UHFFFAOYSA-N 0.000 description 4
- HMRWQTHUDVXMGH-GUBZILKMSA-N Ala-Glu-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN HMRWQTHUDVXMGH-GUBZILKMSA-N 0.000 description 4
- JREOBWLIZLXRIS-GUBZILKMSA-N Asn-Glu-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O JREOBWLIZLXRIS-GUBZILKMSA-N 0.000 description 4
- QXHVOUSPVAWEMX-ZLUOBGJFSA-N Asp-Asp-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O QXHVOUSPVAWEMX-ZLUOBGJFSA-N 0.000 description 4
- 238000011725 BALB/c mouse Methods 0.000 description 4
- XXLBHPPXDUWYAG-XQXXSGGOSA-N Gln-Ala-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XXLBHPPXDUWYAG-XQXXSGGOSA-N 0.000 description 4
- HPCOBEHVEHWREJ-DCAQKATOSA-N Gln-Lys-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O HPCOBEHVEHWREJ-DCAQKATOSA-N 0.000 description 4
- AMSSKPUHBUQBOQ-SRVKXCTJSA-N Leu-Ser-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N AMSSKPUHBUQBOQ-SRVKXCTJSA-N 0.000 description 4
- CLBGMWIYPYAZPR-AVGNSLFASA-N Lys-Arg-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O CLBGMWIYPYAZPR-AVGNSLFASA-N 0.000 description 4
- QLFAPXUXEBAWEK-NHCYSSNCSA-N Lys-Val-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QLFAPXUXEBAWEK-NHCYSSNCSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 108010078144 glutaminyl-glycine Proteins 0.000 description 4
- 230000036039 immunity Effects 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 4
- 230000009257 reactivity Effects 0.000 description 4
- 108010008595 sarcoma-associated antigen S1 Proteins 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 4
- GSCLWXDNIMNIJE-ZLUOBGJFSA-N Ala-Asp-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O GSCLWXDNIMNIJE-ZLUOBGJFSA-N 0.000 description 3
- 108010011667 Ala-Phe-Ala Proteins 0.000 description 3
- SUEIIIFUBHDCCS-PBCZWWQYSA-N Asn-His-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SUEIIIFUBHDCCS-PBCZWWQYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- VZRAXPGTUNDIDK-GUBZILKMSA-N Gln-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N VZRAXPGTUNDIDK-GUBZILKMSA-N 0.000 description 3
- YPMDZWPZFOZYFG-GUBZILKMSA-N Gln-Leu-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YPMDZWPZFOZYFG-GUBZILKMSA-N 0.000 description 3
- PAOHIZNRJNIXQY-XQXXSGGOSA-N Gln-Thr-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O PAOHIZNRJNIXQY-XQXXSGGOSA-N 0.000 description 3
- ZOXBSICWUDAOHX-GUBZILKMSA-N Glu-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O ZOXBSICWUDAOHX-GUBZILKMSA-N 0.000 description 3
- LVCHEMOPBORRLB-DCAQKATOSA-N Glu-Gln-Lys Chemical compound NCCCC[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O LVCHEMOPBORRLB-DCAQKATOSA-N 0.000 description 3
- BUZMZDDKFCSKOT-CIUDSAMLSA-N Glu-Glu-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BUZMZDDKFCSKOT-CIUDSAMLSA-N 0.000 description 3
- SWQALSGKVLYKDT-UHFFFAOYSA-N Gly-Ile-Ala Natural products NCC(=O)NC(C(C)CC)C(=O)NC(C)C(O)=O SWQALSGKVLYKDT-UHFFFAOYSA-N 0.000 description 3
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 3
- 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 3
- ZTLGVASZOIKNIX-DCAQKATOSA-N Leu-Gln-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZTLGVASZOIKNIX-DCAQKATOSA-N 0.000 description 3
- XVZCXCTYGHPNEM-UHFFFAOYSA-N Leu-Leu-Pro Natural products CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(O)=O XVZCXCTYGHPNEM-UHFFFAOYSA-N 0.000 description 3
- HVAUKHLDSDDROB-KKUMJFAQSA-N Lys-Lys-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O HVAUKHLDSDDROB-KKUMJFAQSA-N 0.000 description 3
- 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 3
- 241001529936 Murinae Species 0.000 description 3
- MGBRZXXGQBAULP-DRZSPHRISA-N Phe-Glu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 MGBRZXXGQBAULP-DRZSPHRISA-N 0.000 description 3
- BRKHVZNDAOMAHX-BIIVOSGPSA-N Ser-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N BRKHVZNDAOMAHX-BIIVOSGPSA-N 0.000 description 3
- OHKFXGKHSJKKAL-NRPADANISA-N Ser-Glu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OHKFXGKHSJKKAL-NRPADANISA-N 0.000 description 3
- RRRRCRYTLZVCEN-HJGDQZAQSA-N Thr-Leu-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O RRRRCRYTLZVCEN-HJGDQZAQSA-N 0.000 description 3
- STUAPCLEDMKXKL-LKXGYXEUSA-N Thr-Ser-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O STUAPCLEDMKXKL-LKXGYXEUSA-N 0.000 description 3
- FDKDGFGTHGJKNV-FHWLQOOXSA-N Tyr-Phe-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N FDKDGFGTHGJKNV-FHWLQOOXSA-N 0.000 description 3
- CELJCNRXKZPTCX-XPUUQOCRSA-N Val-Gly-Ala Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O CELJCNRXKZPTCX-XPUUQOCRSA-N 0.000 description 3
- 108010087924 alanylproline Proteins 0.000 description 3
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 3
- 108010068265 aspartyltyrosine Proteins 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000007910 cell fusion Effects 0.000 description 3
- 239000013592 cell lysate Substances 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 238000012790 confirmation Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 108010089804 glycyl-threonine Proteins 0.000 description 3
- 108010050848 glycylleucine Proteins 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 108010012058 leucyltyrosine Proteins 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000001668 nucleic acid synthesis Methods 0.000 description 3
- 239000013600 plasmid vector Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- 238000003118 sandwich ELISA Methods 0.000 description 3
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 3
- 108010026333 seryl-proline Proteins 0.000 description 3
- 229960005486 vaccine Drugs 0.000 description 3
- CWFMWBHMIMNZLN-NAKRPEOUSA-N (2s)-1-[(2s)-2-[[(2s,3s)-2-amino-3-methylpentanoyl]amino]propanoyl]pyrrolidine-2-carboxylic acid Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CWFMWBHMIMNZLN-NAKRPEOUSA-N 0.000 description 2
- OMMDTNGURYRDAC-NRPADANISA-N Ala-Glu-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OMMDTNGURYRDAC-NRPADANISA-N 0.000 description 2
- PNALXAODQKTNLV-JBDRJPRFSA-N Ala-Ile-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O PNALXAODQKTNLV-JBDRJPRFSA-N 0.000 description 2
- GFEDXKNBZMPEDM-KZVJFYERSA-N Ala-Met-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GFEDXKNBZMPEDM-KZVJFYERSA-N 0.000 description 2
- DXQIQUIQYAGRCC-CIUDSAMLSA-N Arg-Asp-Gln Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)CN=C(N)N DXQIQUIQYAGRCC-CIUDSAMLSA-N 0.000 description 2
- DPLFNLDACGGBAK-KKUMJFAQSA-N Arg-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N DPLFNLDACGGBAK-KKUMJFAQSA-N 0.000 description 2
- PRLPSDIHSRITSF-UNQGMJICSA-N Arg-Phe-Thr 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)O)C(O)=O PRLPSDIHSRITSF-UNQGMJICSA-N 0.000 description 2
- DXVMJJNAOVECBA-WHFBIAKZSA-N Asn-Gly-Asn Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O DXVMJJNAOVECBA-WHFBIAKZSA-N 0.000 description 2
- FHETWELNCBMRMG-HJGDQZAQSA-N Asn-Leu-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FHETWELNCBMRMG-HJGDQZAQSA-N 0.000 description 2
- TZFQICWZWFNIKU-KKUMJFAQSA-N Asn-Leu-Tyr Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 TZFQICWZWFNIKU-KKUMJFAQSA-N 0.000 description 2
- KNDCWFXCFKSEBM-AVGNSLFASA-N Asp-Tyr-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O KNDCWFXCFKSEBM-AVGNSLFASA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 description 2
- PHZYLYASFWHLHJ-FXQIFTODSA-N Gln-Asn-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PHZYLYASFWHLHJ-FXQIFTODSA-N 0.000 description 2
- CITDWMLWXNUQKD-FXQIFTODSA-N Gln-Gln-Asn Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N CITDWMLWXNUQKD-FXQIFTODSA-N 0.000 description 2
- VPKBCVUDBNINAH-GARJFASQSA-N Glu-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)O)N)C(=O)O VPKBCVUDBNINAH-GARJFASQSA-N 0.000 description 2
- PVBBEKPHARMPHX-DCAQKATOSA-N Glu-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCC(O)=O PVBBEKPHARMPHX-DCAQKATOSA-N 0.000 description 2
- UMZHHILWZBFPGL-LOKLDPHHSA-N Glu-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O UMZHHILWZBFPGL-LOKLDPHHSA-N 0.000 description 2
- MXXXVOYFNVJHMA-IUCAKERBSA-N Gly-Arg-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CN MXXXVOYFNVJHMA-IUCAKERBSA-N 0.000 description 2
- UPADCCSMVOQAGF-LBPRGKRZSA-N Gly-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)CN)C(O)=O)=CNC2=C1 UPADCCSMVOQAGF-LBPRGKRZSA-N 0.000 description 2
- AAHSHTLISQUZJL-QSFUFRPTSA-N Gly-Ile-Ile Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O AAHSHTLISQUZJL-QSFUFRPTSA-N 0.000 description 2
- IGOYNRWLWHWAQO-JTQLQIEISA-N Gly-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IGOYNRWLWHWAQO-JTQLQIEISA-N 0.000 description 2
- 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 2
- SFOXOSKVTLDEDM-HOTGVXAUSA-N Gly-Trp-Leu Chemical compound C1=CC=C2C(C[C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CN)=CNC2=C1 SFOXOSKVTLDEDM-HOTGVXAUSA-N 0.000 description 2
- RYAOJUMWLWUGNW-QMMMGPOBSA-N Gly-Val-Gly Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O RYAOJUMWLWUGNW-QMMMGPOBSA-N 0.000 description 2
- STOOMQFEJUVAKR-KKUMJFAQSA-N His-His-His Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)C1=CNC=N1 STOOMQFEJUVAKR-KKUMJFAQSA-N 0.000 description 2
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 2
- SAVXZJYTTQQQDD-QEWYBTABSA-N Ile-Phe-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N SAVXZJYTTQQQDD-QEWYBTABSA-N 0.000 description 2
- 241000712431 Influenza A virus Species 0.000 description 2
- QLQHWWCSCLZUMA-KKUMJFAQSA-N Leu-Asp-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QLQHWWCSCLZUMA-KKUMJFAQSA-N 0.000 description 2
- LOLUPZNNADDTAA-AVGNSLFASA-N Leu-Gln-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LOLUPZNNADDTAA-AVGNSLFASA-N 0.000 description 2
- FMEICTQWUKNAGC-YUMQZZPRSA-N Leu-Gly-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O FMEICTQWUKNAGC-YUMQZZPRSA-N 0.000 description 2
- DBSLVQBXKVKDKJ-BJDJZHNGSA-N Leu-Ile-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O DBSLVQBXKVKDKJ-BJDJZHNGSA-N 0.000 description 2
- WXUOJXIGOPMDJM-SRVKXCTJSA-N Leu-Lys-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O WXUOJXIGOPMDJM-SRVKXCTJSA-N 0.000 description 2
- IDGZVZJLYFTXSL-DCAQKATOSA-N Leu-Ser-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IDGZVZJLYFTXSL-DCAQKATOSA-N 0.000 description 2
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 2
- RZHLIPMZXOEJTL-AVGNSLFASA-N Lys-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N RZHLIPMZXOEJTL-AVGNSLFASA-N 0.000 description 2
- QKXZCUCBFPEXNK-KKUMJFAQSA-N Lys-Leu-His Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 QKXZCUCBFPEXNK-KKUMJFAQSA-N 0.000 description 2
- QQPSCXKFDSORFT-IHRRRGAJSA-N Lys-Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN QQPSCXKFDSORFT-IHRRRGAJSA-N 0.000 description 2
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 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 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- BBDSZDHUCPSYAC-QEJZJMRPSA-N Phe-Ala-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BBDSZDHUCPSYAC-QEJZJMRPSA-N 0.000 description 2
- BIYWZVCPZIFGPY-QWRGUYRKSA-N Phe-Gly-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CO)C(O)=O BIYWZVCPZIFGPY-QWRGUYRKSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- QCARZLHECSFOGG-CIUDSAMLSA-N Pro-Glu-Cys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CS)C(=O)O QCARZLHECSFOGG-CIUDSAMLSA-N 0.000 description 2
- ULIWFCCJIOEHMU-BQBZGAKWSA-N Pro-Gly-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 ULIWFCCJIOEHMU-BQBZGAKWSA-N 0.000 description 2
- GURGCNUWVSDYTP-SRVKXCTJSA-N Pro-Leu-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O GURGCNUWVSDYTP-SRVKXCTJSA-N 0.000 description 2
- XYSXOCIWCPFOCG-IHRRRGAJSA-N Pro-Leu-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O XYSXOCIWCPFOCG-IHRRRGAJSA-N 0.000 description 2
- RMODQFBNDDENCP-IHRRRGAJSA-N Pro-Lys-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O RMODQFBNDDENCP-IHRRRGAJSA-N 0.000 description 2
- OWQXAJQZLWHPBH-FXQIFTODSA-N Pro-Ser-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O OWQXAJQZLWHPBH-FXQIFTODSA-N 0.000 description 2
- HQTKVSCNCDLXSX-BQBZGAKWSA-N Ser-Arg-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O HQTKVSCNCDLXSX-BQBZGAKWSA-N 0.000 description 2
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 2
- 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 2
- ILZAUMFXKSIUEF-SRVKXCTJSA-N Ser-Ser-Phe Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 ILZAUMFXKSIUEF-SRVKXCTJSA-N 0.000 description 2
- UBTNVMGPMYDYIU-HJPIBITLSA-N Ser-Tyr-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UBTNVMGPMYDYIU-HJPIBITLSA-N 0.000 description 2
- KIEIJCFVGZCUAS-MELADBBJSA-N Ser-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CO)N)C(=O)O KIEIJCFVGZCUAS-MELADBBJSA-N 0.000 description 2
- HSWXBJCBYSWBPT-GUBZILKMSA-N Ser-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)C(O)=O HSWXBJCBYSWBPT-GUBZILKMSA-N 0.000 description 2
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 2
- GMXIJHCBTZDAPD-QPHKQPEJSA-N Thr-Ile-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N GMXIJHCBTZDAPD-QPHKQPEJSA-N 0.000 description 2
- MEJHFIOYJHTWMK-VOAKCMCISA-N Thr-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)[C@@H](C)O MEJHFIOYJHTWMK-VOAKCMCISA-N 0.000 description 2
- JLNMFGCJODTXDH-WEDXCCLWSA-N Thr-Lys-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O JLNMFGCJODTXDH-WEDXCCLWSA-N 0.000 description 2
- 101710120037 Toxin CcdB Proteins 0.000 description 2
- AKFLVKKWVZMFOT-IHRRRGAJSA-N Tyr-Arg-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O AKFLVKKWVZMFOT-IHRRRGAJSA-N 0.000 description 2
- WVRUKYLYMFGKAN-IHRRRGAJSA-N Tyr-Glu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 WVRUKYLYMFGKAN-IHRRRGAJSA-N 0.000 description 2
- MNWINJDPGBNOED-ULQDDVLXSA-N Tyr-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=C(O)C=C1 MNWINJDPGBNOED-ULQDDVLXSA-N 0.000 description 2
- FZADUTOCSFDBRV-RNXOBYDBSA-N Tyr-Tyr-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=C(O)C=C1 FZADUTOCSFDBRV-RNXOBYDBSA-N 0.000 description 2
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 2
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 2
- CFSSLXZJEMERJY-NRPADANISA-N Val-Gln-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CFSSLXZJEMERJY-NRPADANISA-N 0.000 description 2
- RFKJNTRMXGCKFE-FHWLQOOXSA-N Val-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC(C)C)C(O)=O)=CNC2=C1 RFKJNTRMXGCKFE-FHWLQOOXSA-N 0.000 description 2
- YKNOJPJWNVHORX-UNQGMJICSA-N Val-Phe-Thr Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)O)C(O)=O)CC1=CC=CC=C1 YKNOJPJWNVHORX-UNQGMJICSA-N 0.000 description 2
- 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 2
- 230000006978 adaptation Effects 0.000 description 2
- 238000001042 affinity chromatography Methods 0.000 description 2
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 2
- 229940037003 alum Drugs 0.000 description 2
- 238000011091 antibody purification Methods 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 238000010241 blood sampling Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 2
- 238000002848 electrochemical method Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 2
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 description 2
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 2
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 2
- 108010081551 glycylphenylalanine Proteins 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 239000003547 immunosorbent Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 108010047926 leucyl-lysyl-tyrosine Proteins 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 108010051242 phenylalanylserine Proteins 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 230000012846 protein folding Effects 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 2
- 108010080629 tryptophan-leucine Proteins 0.000 description 2
- 108010038745 tryptophylglycine Proteins 0.000 description 2
- 108010020532 tyrosyl-proline Proteins 0.000 description 2
- GJLXVWOMRRWCIB-MERZOTPQSA-N (2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-acetamido-5-(diaminomethylideneamino)pentanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanamide Chemical compound C([C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(N)=O)C1=CC=C(O)C=C1 GJLXVWOMRRWCIB-MERZOTPQSA-N 0.000 description 1
- XVZCXCTYGHPNEM-IHRRRGAJSA-N (2s)-1-[(2s)-2-[[(2s)-2-amino-4-methylpentanoyl]amino]-4-methylpentanoyl]pyrrolidine-2-carboxylic acid Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(O)=O XVZCXCTYGHPNEM-IHRRRGAJSA-N 0.000 description 1
- DKJPOZOEBONHFS-ZLUOBGJFSA-N Ala-Ala-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O DKJPOZOEBONHFS-ZLUOBGJFSA-N 0.000 description 1
- UWQJHXKARZWDIJ-ZLUOBGJFSA-N Ala-Ala-Cys Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CS)C(O)=O UWQJHXKARZWDIJ-ZLUOBGJFSA-N 0.000 description 1
- LGQPPBQRUBVTIF-JBDRJPRFSA-N Ala-Ala-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LGQPPBQRUBVTIF-JBDRJPRFSA-N 0.000 description 1
- GFBLJMHGHAXGNY-ZLUOBGJFSA-N Ala-Asn-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O GFBLJMHGHAXGNY-ZLUOBGJFSA-N 0.000 description 1
- ZEXDYVGDZJBRMO-ACZMJKKPSA-N Ala-Asn-Gln Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N ZEXDYVGDZJBRMO-ACZMJKKPSA-N 0.000 description 1
- CVGNCMIULZNYES-WHFBIAKZSA-N Ala-Asn-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CVGNCMIULZNYES-WHFBIAKZSA-N 0.000 description 1
- OILNWMNBLIHXQK-ZLUOBGJFSA-N Ala-Cys-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O OILNWMNBLIHXQK-ZLUOBGJFSA-N 0.000 description 1
- NWVVKQZOVSTDBQ-CIUDSAMLSA-N Ala-Glu-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NWVVKQZOVSTDBQ-CIUDSAMLSA-N 0.000 description 1
- WKOBSJOZRJJVRZ-FXQIFTODSA-N Ala-Glu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WKOBSJOZRJJVRZ-FXQIFTODSA-N 0.000 description 1
- PAIHPOGPJVUFJY-WDSKDSINSA-N Ala-Glu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PAIHPOGPJVUFJY-WDSKDSINSA-N 0.000 description 1
- WGDNWOMKBUXFHR-BQBZGAKWSA-N Ala-Gly-Arg Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N WGDNWOMKBUXFHR-BQBZGAKWSA-N 0.000 description 1
- BLIMFWGRQKRCGT-YUMQZZPRSA-N Ala-Gly-Lys Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN BLIMFWGRQKRCGT-YUMQZZPRSA-N 0.000 description 1
- MQIGTEQXYCRLGK-BQBZGAKWSA-N Ala-Gly-Pro Chemical compound C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O MQIGTEQXYCRLGK-BQBZGAKWSA-N 0.000 description 1
- SIGTYDNEPYEXGK-ZANVPECISA-N Ala-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)[C@@H](N)C)C(O)=O)=CNC2=C1 SIGTYDNEPYEXGK-ZANVPECISA-N 0.000 description 1
- HQJKCXHQNUCKMY-GHCJXIJMSA-N Ala-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C)N HQJKCXHQNUCKMY-GHCJXIJMSA-N 0.000 description 1
- MFMDKJIPHSWSBM-GUBZILKMSA-N Ala-Lys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFMDKJIPHSWSBM-GUBZILKMSA-N 0.000 description 1
- PMQXMXAASGFUDX-SRVKXCTJSA-N Ala-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)CCCCN PMQXMXAASGFUDX-SRVKXCTJSA-N 0.000 description 1
- CHFFHQUVXHEGBY-GARJFASQSA-N Ala-Lys-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N CHFFHQUVXHEGBY-GARJFASQSA-N 0.000 description 1
- XRUJOVRWNMBAAA-NHCYSSNCSA-N Ala-Phe-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 XRUJOVRWNMBAAA-NHCYSSNCSA-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
- KLALXKYLOMZDQT-ZLUOBGJFSA-N Ala-Ser-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(N)=O KLALXKYLOMZDQT-ZLUOBGJFSA-N 0.000 description 1
- YYAVDNKUWLAFCV-ACZMJKKPSA-N Ala-Ser-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O YYAVDNKUWLAFCV-ACZMJKKPSA-N 0.000 description 1
- XQNRANMFRPCFFW-GCJQMDKQSA-N Ala-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C)N)O XQNRANMFRPCFFW-GCJQMDKQSA-N 0.000 description 1
- IETUUAHKCHOQHP-KZVJFYERSA-N Ala-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)[C@@H](C)O)C(O)=O IETUUAHKCHOQHP-KZVJFYERSA-N 0.000 description 1
- YEBZNKPPOHFZJM-BPNCWPANSA-N Ala-Tyr-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O YEBZNKPPOHFZJM-BPNCWPANSA-N 0.000 description 1
- BVBKBQRPOJFCQM-DCAQKATOSA-N Arg-Asn-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BVBKBQRPOJFCQM-DCAQKATOSA-N 0.000 description 1
- MZRBYBIQTIKERR-GUBZILKMSA-N Arg-Glu-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MZRBYBIQTIKERR-GUBZILKMSA-N 0.000 description 1
- AGVNTAUPLWIQEN-ZPFDUUQYSA-N Arg-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AGVNTAUPLWIQEN-ZPFDUUQYSA-N 0.000 description 1
- PYZPXCZNQSEHDT-GUBZILKMSA-N Arg-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N PYZPXCZNQSEHDT-GUBZILKMSA-N 0.000 description 1
- FVBZXNSRIDVYJS-AVGNSLFASA-N Arg-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCN=C(N)N FVBZXNSRIDVYJS-AVGNSLFASA-N 0.000 description 1
- FBXMCPLCVYUWBO-BPUTZDHNSA-N Arg-Ser-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)N FBXMCPLCVYUWBO-BPUTZDHNSA-N 0.000 description 1
- AUZAXCPWMDBWEE-HJGDQZAQSA-N Arg-Thr-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O AUZAXCPWMDBWEE-HJGDQZAQSA-N 0.000 description 1
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 1
- NVPHRWNWTKYIST-BPNCWPANSA-N Arg-Tyr-Ala Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 NVPHRWNWTKYIST-BPNCWPANSA-N 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- SWLOHUMCUDRTCL-ZLUOBGJFSA-N Asn-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N SWLOHUMCUDRTCL-ZLUOBGJFSA-N 0.000 description 1
- LEFKSBYHUGUWLP-ACZMJKKPSA-N Asn-Ala-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LEFKSBYHUGUWLP-ACZMJKKPSA-N 0.000 description 1
- SLKLLQWZQHXYSV-CIUDSAMLSA-N Asn-Ala-Lys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O SLKLLQWZQHXYSV-CIUDSAMLSA-N 0.000 description 1
- AKEBUSZTMQLNIX-UWJYBYFXSA-N Asn-Ala-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N AKEBUSZTMQLNIX-UWJYBYFXSA-N 0.000 description 1
- GMRGSBAMMMVDGG-GUBZILKMSA-N Asn-Arg-Arg Chemical compound C(C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)CN=C(N)N GMRGSBAMMMVDGG-GUBZILKMSA-N 0.000 description 1
- BDMIFVIWCNLDCT-CIUDSAMLSA-N Asn-Arg-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O BDMIFVIWCNLDCT-CIUDSAMLSA-N 0.000 description 1
- JZRLLSOWDYUKOK-SRVKXCTJSA-N Asn-Asp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N JZRLLSOWDYUKOK-SRVKXCTJSA-N 0.000 description 1
- XQQVCUIBGYFKDC-OLHMAJIHSA-N Asn-Asp-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XQQVCUIBGYFKDC-OLHMAJIHSA-N 0.000 description 1
- HLTLEIXYIJDFOY-ZLUOBGJFSA-N Asn-Cys-Asn Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O HLTLEIXYIJDFOY-ZLUOBGJFSA-N 0.000 description 1
- SQZIAWGBBUSSPJ-ZKWXMUAHSA-N Asn-Cys-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(=O)N)N SQZIAWGBBUSSPJ-ZKWXMUAHSA-N 0.000 description 1
- XVAPVJNJGLWGCS-ACZMJKKPSA-N Asn-Glu-Asn Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N XVAPVJNJGLWGCS-ACZMJKKPSA-N 0.000 description 1
- MECFLTFREHAZLH-ACZMJKKPSA-N Asn-Glu-Cys Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)N)N MECFLTFREHAZLH-ACZMJKKPSA-N 0.000 description 1
- QYXNFROWLZPWPC-FXQIFTODSA-N Asn-Glu-Gln Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QYXNFROWLZPWPC-FXQIFTODSA-N 0.000 description 1
- PBSQFBAJKPLRJY-BYULHYEWSA-N Asn-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N PBSQFBAJKPLRJY-BYULHYEWSA-N 0.000 description 1
- OOWSBIOUKIUWLO-RCOVLWMOSA-N Asn-Gly-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O OOWSBIOUKIUWLO-RCOVLWMOSA-N 0.000 description 1
- ODBSSLHUFPJRED-CIUDSAMLSA-N Asn-His-Asn Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N ODBSSLHUFPJRED-CIUDSAMLSA-N 0.000 description 1
- NKLRWRRVYGQNIH-GHCJXIJMSA-N Asn-Ile-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O NKLRWRRVYGQNIH-GHCJXIJMSA-N 0.000 description 1
- JQBCANGGAVVERB-CFMVVWHZSA-N Asn-Ile-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N JQBCANGGAVVERB-CFMVVWHZSA-N 0.000 description 1
- SPCONPVIDFMDJI-QSFUFRPTSA-N Asn-Ile-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O SPCONPVIDFMDJI-QSFUFRPTSA-N 0.000 description 1
- ZMUQQMGITUJQTI-CIUDSAMLSA-N Asn-Leu-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O ZMUQQMGITUJQTI-CIUDSAMLSA-N 0.000 description 1
- BZWRLDPIWKOVKB-ZPFDUUQYSA-N Asn-Leu-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BZWRLDPIWKOVKB-ZPFDUUQYSA-N 0.000 description 1
- FODVBOKTYKYRFJ-CIUDSAMLSA-N Asn-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)N)N FODVBOKTYKYRFJ-CIUDSAMLSA-N 0.000 description 1
- LZLCLRQMUQWUHJ-GUBZILKMSA-N Asn-Lys-Gln Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N LZLCLRQMUQWUHJ-GUBZILKMSA-N 0.000 description 1
- ZYPWIUFLYMQZBS-SRVKXCTJSA-N Asn-Lys-Lys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZYPWIUFLYMQZBS-SRVKXCTJSA-N 0.000 description 1
- NLDNNZKUSLAYFW-NHCYSSNCSA-N Asn-Lys-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O NLDNNZKUSLAYFW-NHCYSSNCSA-N 0.000 description 1
- IDUUACUJKUXKKD-VEVYYDQMSA-N Asn-Pro-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O IDUUACUJKUXKKD-VEVYYDQMSA-N 0.000 description 1
- GZXOUBTUAUAVHD-ACZMJKKPSA-N Asn-Ser-Glu Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O GZXOUBTUAUAVHD-ACZMJKKPSA-N 0.000 description 1
- UGXYFDQFLVCDFC-CIUDSAMLSA-N Asn-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O UGXYFDQFLVCDFC-CIUDSAMLSA-N 0.000 description 1
- UXHYOWXTJLBEPG-GSSVUCPTSA-N Asn-Thr-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UXHYOWXTJLBEPG-GSSVUCPTSA-N 0.000 description 1
- RTFXPCYMDYBZNQ-SRVKXCTJSA-N Asn-Tyr-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O RTFXPCYMDYBZNQ-SRVKXCTJSA-N 0.000 description 1
- LRCIOEVFVGXZKB-BZSNNMDCSA-N Asn-Tyr-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O LRCIOEVFVGXZKB-BZSNNMDCSA-N 0.000 description 1
- LMIWYCWRJVMAIQ-NHCYSSNCSA-N Asn-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N LMIWYCWRJVMAIQ-NHCYSSNCSA-N 0.000 description 1
- JGDBHIVECJGXJA-FXQIFTODSA-N Asp-Asp-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JGDBHIVECJGXJA-FXQIFTODSA-N 0.000 description 1
- VPSHHQXIWLGVDD-ZLUOBGJFSA-N Asp-Asp-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O VPSHHQXIWLGVDD-ZLUOBGJFSA-N 0.000 description 1
- FANQWNCPNFEPGZ-WHFBIAKZSA-N Asp-Asp-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O FANQWNCPNFEPGZ-WHFBIAKZSA-N 0.000 description 1
- NYQHSUGFEWDWPD-ACZMJKKPSA-N Asp-Gln-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)O)N NYQHSUGFEWDWPD-ACZMJKKPSA-N 0.000 description 1
- LJRPYAZQQWHEEV-FXQIFTODSA-N Asp-Gln-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O LJRPYAZQQWHEEV-FXQIFTODSA-N 0.000 description 1
- UMHUHHJMEXNSIV-CIUDSAMLSA-N Asp-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UMHUHHJMEXNSIV-CIUDSAMLSA-N 0.000 description 1
- PCJOFZYFFMBZKC-PCBIJLKTSA-N Asp-Phe-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O PCJOFZYFFMBZKC-PCBIJLKTSA-N 0.000 description 1
- RPUYTJJZXQBWDT-SRVKXCTJSA-N Asp-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](CC(=O)O)N RPUYTJJZXQBWDT-SRVKXCTJSA-N 0.000 description 1
- YFGUZQQCSDZRBN-DCAQKATOSA-N Asp-Pro-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YFGUZQQCSDZRBN-DCAQKATOSA-N 0.000 description 1
- KBJVTFWQWXCYCQ-IUKAMOBKSA-N Asp-Thr-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KBJVTFWQWXCYCQ-IUKAMOBKSA-N 0.000 description 1
- ALMIMUZAWTUNIO-BZSNNMDCSA-N Asp-Tyr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ALMIMUZAWTUNIO-BZSNNMDCSA-N 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- DCXGXDGGXVZVMY-GHCJXIJMSA-N Cys-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CS DCXGXDGGXVZVMY-GHCJXIJMSA-N 0.000 description 1
- UUERSUCTHOZPMG-SRVKXCTJSA-N Cys-Asn-Tyr Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 UUERSUCTHOZPMG-SRVKXCTJSA-N 0.000 description 1
- IIGHQOPGMGKDMT-SRVKXCTJSA-N Cys-Asp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CS)N IIGHQOPGMGKDMT-SRVKXCTJSA-N 0.000 description 1
- OZSBRCONEMXYOJ-AVGNSLFASA-N Cys-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CS)N OZSBRCONEMXYOJ-AVGNSLFASA-N 0.000 description 1
- BCFXQBXXDSEHRS-FXQIFTODSA-N Cys-Ser-Arg Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O BCFXQBXXDSEHRS-FXQIFTODSA-N 0.000 description 1
- UEHCDNYDBBCQEL-CIUDSAMLSA-N Cys-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CS)N UEHCDNYDBBCQEL-CIUDSAMLSA-N 0.000 description 1
- CLEFUAZULXANBU-MELADBBJSA-N Cys-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CS)N)C(=O)O CLEFUAZULXANBU-MELADBBJSA-N 0.000 description 1
- VRJZMZGGAKVSIQ-SRVKXCTJSA-N Cys-Tyr-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O VRJZMZGGAKVSIQ-SRVKXCTJSA-N 0.000 description 1
- VIOQRFNAZDMVLO-NRPADANISA-N Cys-Val-Glu Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O VIOQRFNAZDMVLO-NRPADANISA-N 0.000 description 1
- 241000672609 Escherichia coli BL21 Species 0.000 description 1
- HHWQMFIGMMOVFK-WDSKDSINSA-N Gln-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O HHWQMFIGMMOVFK-WDSKDSINSA-N 0.000 description 1
- 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 1
- BVELAHPZLYLZDJ-HGNGGELXSA-N Gln-His-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(O)=O BVELAHPZLYLZDJ-HGNGGELXSA-N 0.000 description 1
- ITZWDGBYBPUZRG-KBIXCLLPSA-N Gln-Ile-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O ITZWDGBYBPUZRG-KBIXCLLPSA-N 0.000 description 1
- CAXXTYYGFYTBPV-IUCAKERBSA-N Gln-Leu-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CAXXTYYGFYTBPV-IUCAKERBSA-N 0.000 description 1
- BJPPYOMRAVLXBY-YUMQZZPRSA-N Gln-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCC(=O)N)N BJPPYOMRAVLXBY-YUMQZZPRSA-N 0.000 description 1
- AQPZYBSRDRZBAG-AVGNSLFASA-N Gln-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N AQPZYBSRDRZBAG-AVGNSLFASA-N 0.000 description 1
- ZGHMRONFHDVXEF-AVGNSLFASA-N Gln-Ser-Phe Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZGHMRONFHDVXEF-AVGNSLFASA-N 0.000 description 1
- SGVGIVDZLSHSEN-RYUDHWBXSA-N Gln-Tyr-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O SGVGIVDZLSHSEN-RYUDHWBXSA-N 0.000 description 1
- UBRQJXFDVZNYJP-AVGNSLFASA-N Gln-Tyr-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O UBRQJXFDVZNYJP-AVGNSLFASA-N 0.000 description 1
- CSMHMEATMDCQNY-DZKIICNBSA-N Gln-Val-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CSMHMEATMDCQNY-DZKIICNBSA-N 0.000 description 1
- SRZLHYPAOXBBSB-HJGDQZAQSA-N Glu-Arg-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SRZLHYPAOXBBSB-HJGDQZAQSA-N 0.000 description 1
- CKRUHITYRFNUKW-WDSKDSINSA-N Glu-Asn-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CKRUHITYRFNUKW-WDSKDSINSA-N 0.000 description 1
- DSPQRJXOIXHOHK-WDSKDSINSA-N Glu-Asp-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O DSPQRJXOIXHOHK-WDSKDSINSA-N 0.000 description 1
- OBIHEDRRSMRKLU-ACZMJKKPSA-N Glu-Cys-Asp Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(=O)O)C(=O)O)N OBIHEDRRSMRKLU-ACZMJKKPSA-N 0.000 description 1
- OXEMJGCAJFFREE-FXQIFTODSA-N Glu-Gln-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O OXEMJGCAJFFREE-FXQIFTODSA-N 0.000 description 1
- IQACOVZVOMVILH-FXQIFTODSA-N Glu-Glu-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O IQACOVZVOMVILH-FXQIFTODSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- LRPXYSGPOBVBEH-IUCAKERBSA-N Glu-Gly-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O LRPXYSGPOBVBEH-IUCAKERBSA-N 0.000 description 1
- RAUDKMVXNOWDLS-WDSKDSINSA-N Glu-Gly-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O RAUDKMVXNOWDLS-WDSKDSINSA-N 0.000 description 1
- HILMIYALTUQTRC-XVKPBYJWSA-N Glu-Gly-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HILMIYALTUQTRC-XVKPBYJWSA-N 0.000 description 1
- ZWABFSSWTSAMQN-KBIXCLLPSA-N Glu-Ile-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O ZWABFSSWTSAMQN-KBIXCLLPSA-N 0.000 description 1
- LGYCLOCORAEQSZ-PEFMBERDSA-N Glu-Ile-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O LGYCLOCORAEQSZ-PEFMBERDSA-N 0.000 description 1
- VSRCAOIHMGCIJK-SRVKXCTJSA-N Glu-Leu-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O VSRCAOIHMGCIJK-SRVKXCTJSA-N 0.000 description 1
- BCYGDJXHAGZNPQ-DCAQKATOSA-N Glu-Lys-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O BCYGDJXHAGZNPQ-DCAQKATOSA-N 0.000 description 1
- ILWHFUZZCFYSKT-AVGNSLFASA-N Glu-Lys-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O ILWHFUZZCFYSKT-AVGNSLFASA-N 0.000 description 1
- HRBYTAIBKPNZKQ-AVGNSLFASA-N Glu-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(O)=O HRBYTAIBKPNZKQ-AVGNSLFASA-N 0.000 description 1
- PMSMKNYRZCKVMC-DRZSPHRISA-N Glu-Phe-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CCC(=O)O)N PMSMKNYRZCKVMC-DRZSPHRISA-N 0.000 description 1
- UERORLSAFUHDGU-AVGNSLFASA-N Glu-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N UERORLSAFUHDGU-AVGNSLFASA-N 0.000 description 1
- BFEZQZKEPRKKHV-SRVKXCTJSA-N Glu-Pro-Lys Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)O)N)C(=O)N[C@@H](CCCCN)C(=O)O BFEZQZKEPRKKHV-SRVKXCTJSA-N 0.000 description 1
- IDEODOAVGCMUQV-GUBZILKMSA-N Glu-Ser-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IDEODOAVGCMUQV-GUBZILKMSA-N 0.000 description 1
- VNCNWQPIQYAMAK-ACZMJKKPSA-N Glu-Ser-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O VNCNWQPIQYAMAK-ACZMJKKPSA-N 0.000 description 1
- HBMRTXJZQDVRFT-DZKIICNBSA-N Glu-Tyr-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O HBMRTXJZQDVRFT-DZKIICNBSA-N 0.000 description 1
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 1
- JVWPPCWUDRJGAE-YUMQZZPRSA-N Gly-Asn-Leu Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O JVWPPCWUDRJGAE-YUMQZZPRSA-N 0.000 description 1
- XRTDOIOIBMAXCT-NKWVEPMBSA-N Gly-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)CN)C(=O)O XRTDOIOIBMAXCT-NKWVEPMBSA-N 0.000 description 1
- RPLLQZBOVIVGMX-QWRGUYRKSA-N Gly-Asp-Phe Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O RPLLQZBOVIVGMX-QWRGUYRKSA-N 0.000 description 1
- PMNHJLASAAWELO-FOHZUACHSA-N Gly-Asp-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PMNHJLASAAWELO-FOHZUACHSA-N 0.000 description 1
- QGZSAHIZRQHCEQ-QWRGUYRKSA-N Gly-Asp-Tyr Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QGZSAHIZRQHCEQ-QWRGUYRKSA-N 0.000 description 1
- HDNXXTBKOJKWNN-WDSKDSINSA-N Gly-Glu-Asn Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O HDNXXTBKOJKWNN-WDSKDSINSA-N 0.000 description 1
- SWQALSGKVLYKDT-ZKWXMUAHSA-N Gly-Ile-Ala Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O SWQALSGKVLYKDT-ZKWXMUAHSA-N 0.000 description 1
- LUJVWKKYHSLULQ-ZKWXMUAHSA-N Gly-Ile-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN LUJVWKKYHSLULQ-ZKWXMUAHSA-N 0.000 description 1
- DENRBIYENOKSEX-PEXQALLHSA-N Gly-Ile-His Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 DENRBIYENOKSEX-PEXQALLHSA-N 0.000 description 1
- VLIJYPMATZSOLL-YUMQZZPRSA-N Gly-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN VLIJYPMATZSOLL-YUMQZZPRSA-N 0.000 description 1
- GMTXWRIDLGTVFC-IUCAKERBSA-N Gly-Lys-Glu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O GMTXWRIDLGTVFC-IUCAKERBSA-N 0.000 description 1
- NTBOEZICHOSJEE-YUMQZZPRSA-N Gly-Lys-Ser Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O NTBOEZICHOSJEE-YUMQZZPRSA-N 0.000 description 1
- JJGBXTYGTKWGAT-YUMQZZPRSA-N Gly-Pro-Glu Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O JJGBXTYGTKWGAT-YUMQZZPRSA-N 0.000 description 1
- 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 1
- LCRDMSSAKLTKBU-ZDLURKLDSA-N Gly-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN LCRDMSSAKLTKBU-ZDLURKLDSA-N 0.000 description 1
- FKYQEVBRZSFAMJ-QWRGUYRKSA-N Gly-Ser-Tyr Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FKYQEVBRZSFAMJ-QWRGUYRKSA-N 0.000 description 1
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-N 0.000 description 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
- MYXNLWDWWOTERK-BHNWBGBOSA-N Gly-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN)O MYXNLWDWWOTERK-BHNWBGBOSA-N 0.000 description 1
- FNXSYBOHALPRHV-ONGXEEELSA-N Gly-Val-Lys Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN FNXSYBOHALPRHV-ONGXEEELSA-N 0.000 description 1
- IZVICCORZOSGPT-JSGCOSHPSA-N Gly-Val-Tyr Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IZVICCORZOSGPT-JSGCOSHPSA-N 0.000 description 1
- DCRODRAURLJOFY-XPUUQOCRSA-N His-Ala-Gly Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)NCC(O)=O DCRODRAURLJOFY-XPUUQOCRSA-N 0.000 description 1
- AWASVTXPTOLPPP-MBLNEYKQSA-N His-Ala-Thr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AWASVTXPTOLPPP-MBLNEYKQSA-N 0.000 description 1
- DFHVLUKTTVTCKY-PBCZWWQYSA-N His-Asn-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N)O DFHVLUKTTVTCKY-PBCZWWQYSA-N 0.000 description 1
- VOEGKUNRHYKYSU-XVYDVKMFSA-N His-Asp-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O VOEGKUNRHYKYSU-XVYDVKMFSA-N 0.000 description 1
- IDQNVIWPPWAFSY-AVGNSLFASA-N His-His-Gln Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(N)=O)C(O)=O IDQNVIWPPWAFSY-AVGNSLFASA-N 0.000 description 1
- YEKYGQZUBCRNGH-DCAQKATOSA-N His-Pro-Ser Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CN=CN2)N)C(=O)N[C@@H](CO)C(=O)O YEKYGQZUBCRNGH-DCAQKATOSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000617823 Homo sapiens Solute carrier organic anion transporter family member 6A1 Proteins 0.000 description 1
- JRHFQUPIZOYKQP-KBIXCLLPSA-N Ile-Ala-Glu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O JRHFQUPIZOYKQP-KBIXCLLPSA-N 0.000 description 1
- AQCUAZTZSPQJFF-ZKWXMUAHSA-N Ile-Ala-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O AQCUAZTZSPQJFF-ZKWXMUAHSA-N 0.000 description 1
- VCYVLFAWCJRXFT-HJPIBITLSA-N Ile-Cys-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N VCYVLFAWCJRXFT-HJPIBITLSA-N 0.000 description 1
- SLQVFYWBGNNOTK-BYULHYEWSA-N Ile-Gly-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)N)C(=O)O)N SLQVFYWBGNNOTK-BYULHYEWSA-N 0.000 description 1
- AXNGDPAKKCEKGY-QPHKQPEJSA-N Ile-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N AXNGDPAKKCEKGY-QPHKQPEJSA-N 0.000 description 1
- KLBVGHCGHUNHEA-BJDJZHNGSA-N Ile-Leu-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)O)N KLBVGHCGHUNHEA-BJDJZHNGSA-N 0.000 description 1
- OUUCIIJSBIBCHB-ZPFDUUQYSA-N Ile-Leu-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O OUUCIIJSBIBCHB-ZPFDUUQYSA-N 0.000 description 1
- TVYWVSJGSHQWMT-AJNGGQMLSA-N Ile-Leu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N TVYWVSJGSHQWMT-AJNGGQMLSA-N 0.000 description 1
- FHPZJWJWTWZKNA-LLLHUVSDSA-N Ile-Phe-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N FHPZJWJWTWZKNA-LLLHUVSDSA-N 0.000 description 1
- XMYURPUVJSKTMC-KBIXCLLPSA-N Ile-Ser-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N XMYURPUVJSKTMC-KBIXCLLPSA-N 0.000 description 1
- QGXQHJQPAPMACW-PPCPHDFISA-N Ile-Thr-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)O)N QGXQHJQPAPMACW-PPCPHDFISA-N 0.000 description 1
- WCNWGAUZWWSYDG-SVSWQMSJSA-N Ile-Thr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)O)N WCNWGAUZWWSYDG-SVSWQMSJSA-N 0.000 description 1
- QHUREMVLLMNUAX-OSUNSFLBSA-N Ile-Thr-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)O)N QHUREMVLLMNUAX-OSUNSFLBSA-N 0.000 description 1
- GNXGAVNTVNOCLL-SIUGBPQLSA-N Ile-Tyr-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N GNXGAVNTVNOCLL-SIUGBPQLSA-N 0.000 description 1
- YWCJXQKATPNPOE-UKJIMTQDSA-N Ile-Val-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N YWCJXQKATPNPOE-UKJIMTQDSA-N 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- PWWVAXIEGOYWEE-UHFFFAOYSA-N Isophenergan Chemical compound C1=CC=C2N(CC(C)N(C)C)C3=CC=CC=C3SC2=C1 PWWVAXIEGOYWEE-UHFFFAOYSA-N 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- HASRFYOMVPJRPU-SRVKXCTJSA-N Leu-Arg-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O HASRFYOMVPJRPU-SRVKXCTJSA-N 0.000 description 1
- KTFHTMHHKXUYPW-ZPFDUUQYSA-N Leu-Asp-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KTFHTMHHKXUYPW-ZPFDUUQYSA-N 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
- QVFGXCVIXXBFHO-AVGNSLFASA-N Leu-Glu-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O QVFGXCVIXXBFHO-AVGNSLFASA-N 0.000 description 1
- HQUXQAMSWFIRET-AVGNSLFASA-N Leu-Glu-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN HQUXQAMSWFIRET-AVGNSLFASA-N 0.000 description 1
- WQWSMEOYXJTFRU-GUBZILKMSA-N Leu-Glu-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O WQWSMEOYXJTFRU-GUBZILKMSA-N 0.000 description 1
- APFJUBGRZGMQFF-QWRGUYRKSA-N Leu-Gly-Lys Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN APFJUBGRZGMQFF-QWRGUYRKSA-N 0.000 description 1
- AOFYPTOHESIBFZ-KKUMJFAQSA-N Leu-His-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O AOFYPTOHESIBFZ-KKUMJFAQSA-N 0.000 description 1
- KUIDCYNIEJBZBU-AJNGGQMLSA-N Leu-Ile-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O KUIDCYNIEJBZBU-AJNGGQMLSA-N 0.000 description 1
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 1
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 1
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 1
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 description 1
- FOBUGKUBUJOWAD-IHPCNDPISA-N Leu-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FOBUGKUBUJOWAD-IHPCNDPISA-N 0.000 description 1
- JLWZLIQRYCTYBD-IHRRRGAJSA-N Leu-Lys-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JLWZLIQRYCTYBD-IHRRRGAJSA-N 0.000 description 1
- RTIRBWJPYJYTLO-MELADBBJSA-N Leu-Lys-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N RTIRBWJPYJYTLO-MELADBBJSA-N 0.000 description 1
- VCHVSKNMTXWIIP-SRVKXCTJSA-N Leu-Lys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O VCHVSKNMTXWIIP-SRVKXCTJSA-N 0.000 description 1
- ONPJGOIVICHWBW-BZSNNMDCSA-N Leu-Lys-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 ONPJGOIVICHWBW-BZSNNMDCSA-N 0.000 description 1
- INCJJHQRZGQLFC-KBPBESRZSA-N Leu-Phe-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O INCJJHQRZGQLFC-KBPBESRZSA-N 0.000 description 1
- JDBQSGMJBMPNFT-AVGNSLFASA-N Leu-Pro-Val Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O JDBQSGMJBMPNFT-AVGNSLFASA-N 0.000 description 1
- IZPVWNSAVUQBGP-CIUDSAMLSA-N Leu-Ser-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IZPVWNSAVUQBGP-CIUDSAMLSA-N 0.000 description 1
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-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
- GOFJOGXGMPHOGL-DCAQKATOSA-N Leu-Ser-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(C)C GOFJOGXGMPHOGL-DCAQKATOSA-N 0.000 description 1
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 1
- 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
- LFSQWRSVPNKJGP-WDCWCFNPSA-N Leu-Thr-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(O)=O LFSQWRSVPNKJGP-WDCWCFNPSA-N 0.000 description 1
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 1
- FPFOYSCDUWTZBF-IHPCNDPISA-N Leu-Trp-Leu Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H]([NH3+])CC(C)C)C(=O)N[C@@H](CC(C)C)C([O-])=O)=CNC2=C1 FPFOYSCDUWTZBF-IHPCNDPISA-N 0.000 description 1
- RDFIVFHPOSOXMW-ACRUOGEOSA-N Leu-Tyr-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O RDFIVFHPOSOXMW-ACRUOGEOSA-N 0.000 description 1
- XZNJZXJZBMBGGS-NHCYSSNCSA-N Leu-Val-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XZNJZXJZBMBGGS-NHCYSSNCSA-N 0.000 description 1
- YNNPKXBBRZVIRX-IHRRRGAJSA-N Lys-Arg-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O YNNPKXBBRZVIRX-IHRRRGAJSA-N 0.000 description 1
- ABHIXYDMILIUKV-CIUDSAMLSA-N Lys-Asn-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ABHIXYDMILIUKV-CIUDSAMLSA-N 0.000 description 1
- QYOXSYXPHUHOJR-GUBZILKMSA-N Lys-Asn-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O QYOXSYXPHUHOJR-GUBZILKMSA-N 0.000 description 1
- HQVDJTYKCMIWJP-YUMQZZPRSA-N Lys-Asn-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O HQVDJTYKCMIWJP-YUMQZZPRSA-N 0.000 description 1
- DGWXCIORNLWGGG-CIUDSAMLSA-N Lys-Asn-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O DGWXCIORNLWGGG-CIUDSAMLSA-N 0.000 description 1
- XTONYTDATVADQH-CIUDSAMLSA-N Lys-Cys-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O XTONYTDATVADQH-CIUDSAMLSA-N 0.000 description 1
- RDIILCRAWOSDOQ-CIUDSAMLSA-N Lys-Cys-Asp Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(=O)O)C(=O)O)N RDIILCRAWOSDOQ-CIUDSAMLSA-N 0.000 description 1
- QQUJSUFWEDZQQY-AVGNSLFASA-N Lys-Gln-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CCCCN QQUJSUFWEDZQQY-AVGNSLFASA-N 0.000 description 1
- GJJQCBVRWDGLMQ-GUBZILKMSA-N Lys-Glu-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O GJJQCBVRWDGLMQ-GUBZILKMSA-N 0.000 description 1
- GCMWRRQAKQXDED-IUCAKERBSA-N Lys-Glu-Gly Chemical compound [NH3+]CCCC[C@H]([NH3+])C(=O)N[C@@H](CCC([O-])=O)C(=O)NCC([O-])=O GCMWRRQAKQXDED-IUCAKERBSA-N 0.000 description 1
- VEGLGAOVLFODGC-GUBZILKMSA-N Lys-Glu-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O VEGLGAOVLFODGC-GUBZILKMSA-N 0.000 description 1
- ULUQBUKAPDUKOC-GVXVVHGQSA-N Lys-Glu-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O ULUQBUKAPDUKOC-GVXVVHGQSA-N 0.000 description 1
- NKKFVJRLCCUJNA-QWRGUYRKSA-N Lys-Gly-Lys Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN NKKFVJRLCCUJNA-QWRGUYRKSA-N 0.000 description 1
- HAUUXTXKJNVIFY-ONGXEEELSA-N Lys-Gly-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HAUUXTXKJNVIFY-ONGXEEELSA-N 0.000 description 1
- SLQJJFAVWSZLBL-BJDJZHNGSA-N Lys-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCCCN SLQJJFAVWSZLBL-BJDJZHNGSA-N 0.000 description 1
- NCZIQZYZPUPMKY-PPCPHDFISA-N Lys-Ile-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NCZIQZYZPUPMKY-PPCPHDFISA-N 0.000 description 1
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 1
- AIRZWUMAHCDDHR-KKUMJFAQSA-N Lys-Leu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O AIRZWUMAHCDDHR-KKUMJFAQSA-N 0.000 description 1
- WRODMZBHNNPRLN-SRVKXCTJSA-N Lys-Leu-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O WRODMZBHNNPRLN-SRVKXCTJSA-N 0.000 description 1
- YUAXTFMFMOIMAM-QWRGUYRKSA-N Lys-Lys-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O YUAXTFMFMOIMAM-QWRGUYRKSA-N 0.000 description 1
- HYSVGEAWTGPMOA-IHRRRGAJSA-N Lys-Pro-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O HYSVGEAWTGPMOA-IHRRRGAJSA-N 0.000 description 1
- MGKFCQFVPKOWOL-CIUDSAMLSA-N Lys-Ser-Asp Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N MGKFCQFVPKOWOL-CIUDSAMLSA-N 0.000 description 1
- DIBZLYZXTSVGLN-CIUDSAMLSA-N Lys-Ser-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O DIBZLYZXTSVGLN-CIUDSAMLSA-N 0.000 description 1
- SQRLLZAQNOQCEG-KKUMJFAQSA-N Lys-Tyr-Ser Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CC1=CC=C(O)C=C1 SQRLLZAQNOQCEG-KKUMJFAQSA-N 0.000 description 1
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 1
- HKRYNJSKVLZIFP-IHRRRGAJSA-N Met-Asn-Tyr Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O HKRYNJSKVLZIFP-IHRRRGAJSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 101001028244 Onchocerca volvulus Fatty-acid and retinol-binding protein 1 Proteins 0.000 description 1
- 241000845082 Panama Species 0.000 description 1
- LXVFHIBXOWJTKZ-BZSNNMDCSA-N Phe-Asn-Tyr Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O LXVFHIBXOWJTKZ-BZSNNMDCSA-N 0.000 description 1
- LLGTYVHITPVGKR-RYUDHWBXSA-N Phe-Gln-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O LLGTYVHITPVGKR-RYUDHWBXSA-N 0.000 description 1
- HOYQLNNGMHXZDW-KKUMJFAQSA-N Phe-Glu-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O HOYQLNNGMHXZDW-KKUMJFAQSA-N 0.000 description 1
- MGECUMGTSHYHEJ-QEWYBTABSA-N Phe-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 MGECUMGTSHYHEJ-QEWYBTABSA-N 0.000 description 1
- HGNGAMWHGGANAU-WHOFXGATSA-N Phe-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 HGNGAMWHGGANAU-WHOFXGATSA-N 0.000 description 1
- GYEPCBNTTRORKW-PCBIJLKTSA-N Phe-Ile-Asp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O GYEPCBNTTRORKW-PCBIJLKTSA-N 0.000 description 1
- KRYSMKKRRRWOCZ-QEWYBTABSA-N Phe-Ile-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O KRYSMKKRRRWOCZ-QEWYBTABSA-N 0.000 description 1
- DMEYUTSDVRCWRS-ULQDDVLXSA-N Phe-Lys-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 DMEYUTSDVRCWRS-ULQDDVLXSA-N 0.000 description 1
- ZVRJWDUPIDMHDN-ULQDDVLXSA-N Phe-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=CC=C1 ZVRJWDUPIDMHDN-ULQDDVLXSA-N 0.000 description 1
- MVIJMIZJPHQGEN-IHRRRGAJSA-N Phe-Ser-Val Chemical compound CC(C)[C@@H](C([O-])=O)NC(=O)[C@H](CO)NC(=O)[C@@H]([NH3+])CC1=CC=CC=C1 MVIJMIZJPHQGEN-IHRRRGAJSA-N 0.000 description 1
- YDUGVDGFKNXFPL-IXOXFDKPSA-N Phe-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N)O YDUGVDGFKNXFPL-IXOXFDKPSA-N 0.000 description 1
- SHUFSZDAIPLZLF-BEAPCOKYSA-N Phe-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)O SHUFSZDAIPLZLF-BEAPCOKYSA-N 0.000 description 1
- NHHZWPNMYQUNEH-ACRUOGEOSA-N Phe-Tyr-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CC3=CN=CN3)C(=O)O)N NHHZWPNMYQUNEH-ACRUOGEOSA-N 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- OYEUSRAZOGIDBY-JYJNAYRXSA-N Pro-Arg-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OYEUSRAZOGIDBY-JYJNAYRXSA-N 0.000 description 1
- SBYVDRLQAGENMY-DCAQKATOSA-N Pro-Asn-His Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O SBYVDRLQAGENMY-DCAQKATOSA-N 0.000 description 1
- OZAPWFHRPINHND-GUBZILKMSA-N Pro-Cys-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O OZAPWFHRPINHND-GUBZILKMSA-N 0.000 description 1
- HJSCRFZVGXAGNG-SRVKXCTJSA-N Pro-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 HJSCRFZVGXAGNG-SRVKXCTJSA-N 0.000 description 1
- UAYHMOIGIQZLFR-NHCYSSNCSA-N Pro-Gln-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O UAYHMOIGIQZLFR-NHCYSSNCSA-N 0.000 description 1
- WVOXLKUUVCCCSU-ZPFDUUQYSA-N Pro-Glu-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WVOXLKUUVCCCSU-ZPFDUUQYSA-N 0.000 description 1
- FDINZVJXLPILKV-DCAQKATOSA-N Pro-His-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O FDINZVJXLPILKV-DCAQKATOSA-N 0.000 description 1
- UREQLMJCKFLLHM-NAKRPEOUSA-N Pro-Ile-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O UREQLMJCKFLLHM-NAKRPEOUSA-N 0.000 description 1
- HFNPOYOKIPGAEI-SRVKXCTJSA-N Pro-Leu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 HFNPOYOKIPGAEI-SRVKXCTJSA-N 0.000 description 1
- XQPHBAKJJJZOBX-SRVKXCTJSA-N Pro-Lys-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O XQPHBAKJJJZOBX-SRVKXCTJSA-N 0.000 description 1
- WOIFYRZPIORBRY-AVGNSLFASA-N Pro-Lys-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O WOIFYRZPIORBRY-AVGNSLFASA-N 0.000 description 1
- JFBJPBZSTMXGKL-JYJNAYRXSA-N Pro-Met-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JFBJPBZSTMXGKL-JYJNAYRXSA-N 0.000 description 1
- BGWKULMLUIUPKY-BQBZGAKWSA-N Pro-Ser-Gly Chemical compound OC(=O)CNC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 BGWKULMLUIUPKY-BQBZGAKWSA-N 0.000 description 1
- KWMZPPWYBVZIER-XGEHTFHBSA-N Pro-Ser-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWMZPPWYBVZIER-XGEHTFHBSA-N 0.000 description 1
- WWXNZNWZNZPDIF-SRVKXCTJSA-N Pro-Val-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 WWXNZNWZNZPDIF-SRVKXCTJSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- WDXYVIIVDIDOSX-DCAQKATOSA-N Ser-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N WDXYVIIVDIDOSX-DCAQKATOSA-N 0.000 description 1
- XVAUJOAYHWWNQF-ZLUOBGJFSA-N Ser-Asn-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O XVAUJOAYHWWNQF-ZLUOBGJFSA-N 0.000 description 1
- UCXDHBORXLVBNC-ZLUOBGJFSA-N Ser-Asn-Cys Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(O)=O UCXDHBORXLVBNC-ZLUOBGJFSA-N 0.000 description 1
- DKKGAAJTDKHWOD-BIIVOSGPSA-N Ser-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CO)N)C(=O)O DKKGAAJTDKHWOD-BIIVOSGPSA-N 0.000 description 1
- ICHZYBVODUVUKN-SRVKXCTJSA-N Ser-Asn-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ICHZYBVODUVUKN-SRVKXCTJSA-N 0.000 description 1
- CRZRTKAVUUGKEQ-ACZMJKKPSA-N Ser-Gln-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CRZRTKAVUUGKEQ-ACZMJKKPSA-N 0.000 description 1
- YPUSXTWURJANKF-KBIXCLLPSA-N Ser-Gln-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YPUSXTWURJANKF-KBIXCLLPSA-N 0.000 description 1
- SQBLRDDJTUJDMV-ACZMJKKPSA-N Ser-Glu-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQBLRDDJTUJDMV-ACZMJKKPSA-N 0.000 description 1
- UFKPDBLKLOBMRH-XHNCKOQMSA-N Ser-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N)C(=O)O UFKPDBLKLOBMRH-XHNCKOQMSA-N 0.000 description 1
- MIJWOJAXARLEHA-WDSKDSINSA-N Ser-Gly-Glu Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O MIJWOJAXARLEHA-WDSKDSINSA-N 0.000 description 1
- IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 description 1
- UIGMAMGZOJVTDN-WHFBIAKZSA-N Ser-Gly-Ser Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O UIGMAMGZOJVTDN-WHFBIAKZSA-N 0.000 description 1
- FYUIFUJFNCLUIX-XVYDVKMFSA-N Ser-His-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(O)=O FYUIFUJFNCLUIX-XVYDVKMFSA-N 0.000 description 1
- DOSZISJPMCYEHT-NAKRPEOUSA-N Ser-Ile-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O DOSZISJPMCYEHT-NAKRPEOUSA-N 0.000 description 1
- HEUVHBXOVZONPU-BJDJZHNGSA-N Ser-Leu-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HEUVHBXOVZONPU-BJDJZHNGSA-N 0.000 description 1
- VZQRNAYURWAEFE-KKUMJFAQSA-N Ser-Leu-Phe Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 VZQRNAYURWAEFE-KKUMJFAQSA-N 0.000 description 1
- IFLVBVIYADZIQO-DCAQKATOSA-N Ser-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N IFLVBVIYADZIQO-DCAQKATOSA-N 0.000 description 1
- BUYHXYIUQUBEQP-AVGNSLFASA-N Ser-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CO)N BUYHXYIUQUBEQP-AVGNSLFASA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- ZKBKUWQVDWWSRI-BZSNNMDCSA-N Ser-Phe-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZKBKUWQVDWWSRI-BZSNNMDCSA-N 0.000 description 1
- 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 1
- HHJFMHQYEAAOBM-ZLUOBGJFSA-N Ser-Ser-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O HHJFMHQYEAAOBM-ZLUOBGJFSA-N 0.000 description 1
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 1
- DKGRNFUXVTYRAS-UBHSHLNASA-N Ser-Ser-Trp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O DKGRNFUXVTYRAS-UBHSHLNASA-N 0.000 description 1
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 1
- BDMWLJLPPUCLNV-XGEHTFHBSA-N Ser-Thr-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BDMWLJLPPUCLNV-XGEHTFHBSA-N 0.000 description 1
- YXEYTHXDRDAIOJ-CWRNSKLLSA-N Ser-Trp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CO)N)C(=O)O YXEYTHXDRDAIOJ-CWRNSKLLSA-N 0.000 description 1
- ZWSZBWAFDZRBNM-UBHSHLNASA-N Ser-Trp-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CO)C(O)=O ZWSZBWAFDZRBNM-UBHSHLNASA-N 0.000 description 1
- HAYADTTXNZFUDM-IHRRRGAJSA-N Ser-Tyr-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O HAYADTTXNZFUDM-IHRRRGAJSA-N 0.000 description 1
- PCMZJFMUYWIERL-ZKWXMUAHSA-N Ser-Val-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PCMZJFMUYWIERL-ZKWXMUAHSA-N 0.000 description 1
- SGZVZUCRAVSPKQ-FXQIFTODSA-N Ser-Val-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CO)N SGZVZUCRAVSPKQ-FXQIFTODSA-N 0.000 description 1
- 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 1
- OJRNZRROAIAHDL-LKXGYXEUSA-N Thr-Asn-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OJRNZRROAIAHDL-LKXGYXEUSA-N 0.000 description 1
- XDARBNMYXKUFOJ-GSSVUCPTSA-N Thr-Asp-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XDARBNMYXKUFOJ-GSSVUCPTSA-N 0.000 description 1
- ASJDFGOPDCVXTG-KATARQTJSA-N Thr-Cys-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(O)=O ASJDFGOPDCVXTG-KATARQTJSA-N 0.000 description 1
- MMTOHPRBJKEZHT-BWBBJGPYSA-N Thr-Cys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O MMTOHPRBJKEZHT-BWBBJGPYSA-N 0.000 description 1
- GCXFWAZRHBRYEM-NUMRIWBASA-N Thr-Gln-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O GCXFWAZRHBRYEM-NUMRIWBASA-N 0.000 description 1
- HJOSVGCWOTYJFG-WDCWCFNPSA-N Thr-Glu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N)O HJOSVGCWOTYJFG-WDCWCFNPSA-N 0.000 description 1
- BNGDYRRHRGOPHX-IFFSRLJSSA-N Thr-Glu-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O BNGDYRRHRGOPHX-IFFSRLJSSA-N 0.000 description 1
- 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 1
- GXUWHVZYDAHFSV-FLBSBUHZSA-N Thr-Ile-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GXUWHVZYDAHFSV-FLBSBUHZSA-N 0.000 description 1
- UUSQVWOVUYMLJA-PPCPHDFISA-N Thr-Lys-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UUSQVWOVUYMLJA-PPCPHDFISA-N 0.000 description 1
- SPVHQURZJCUDQC-VOAKCMCISA-N Thr-Lys-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O SPVHQURZJCUDQC-VOAKCMCISA-N 0.000 description 1
- NYQIZWROIMIQSL-VEVYYDQMSA-N Thr-Pro-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O NYQIZWROIMIQSL-VEVYYDQMSA-N 0.000 description 1
- MROIJTGJGIDEEJ-RCWTZXSCSA-N Thr-Pro-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 MROIJTGJGIDEEJ-RCWTZXSCSA-N 0.000 description 1
- YRJOLUDFVAUXLI-GSSVUCPTSA-N Thr-Thr-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O YRJOLUDFVAUXLI-GSSVUCPTSA-N 0.000 description 1
- BZTSQFWJNJYZSX-JRQIVUDYSA-N Thr-Tyr-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O BZTSQFWJNJYZSX-JRQIVUDYSA-N 0.000 description 1
- QNXZCKMXHPULME-ZNSHCXBVSA-N Thr-Val-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O QNXZCKMXHPULME-ZNSHCXBVSA-N 0.000 description 1
- BTAJAOWZCWOHBU-HSHDSVGOSA-N Thr-Val-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)O)C(C)C)C(O)=O)=CNC2=C1 BTAJAOWZCWOHBU-HSHDSVGOSA-N 0.000 description 1
- NIWAGRRZHCMPOY-GMVOTWDCSA-N Trp-Ala-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N NIWAGRRZHCMPOY-GMVOTWDCSA-N 0.000 description 1
- DVIIYMVCSUQOJG-QEJZJMRPSA-N Trp-Glu-Asp Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O DVIIYMVCSUQOJG-QEJZJMRPSA-N 0.000 description 1
- YNJBFSFTYAPMTA-FNINZLGVSA-N Trp-Gly-Ile-His-His Chemical compound CCC(C)[C@H](NC(=O)CNC(=O)[C@@H](N)Cc1c[nH]c2ccccc12)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O YNJBFSFTYAPMTA-FNINZLGVSA-N 0.000 description 1
- CCZXBOFIBYQLEV-IHPCNDPISA-N Trp-Leu-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)Cc1c[nH]c2ccccc12)C(O)=O CCZXBOFIBYQLEV-IHPCNDPISA-N 0.000 description 1
- UKWSFUSPGPBJGU-VFAJRCTISA-N Trp-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O UKWSFUSPGPBJGU-VFAJRCTISA-N 0.000 description 1
- KOVPHHXMHLFWPL-BPUTZDHNSA-N Trp-Pro-Asn Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)N[C@@H](CC(=O)N)C(=O)O KOVPHHXMHLFWPL-BPUTZDHNSA-N 0.000 description 1
- ABRICLFKFRFDKS-IHPCNDPISA-N Trp-Ser-Tyr Chemical compound C([C@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)C(O)=O)C1=CC=C(O)C=C1 ABRICLFKFRFDKS-IHPCNDPISA-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
- ZKVANNIVSDOQMG-HKUYNNGSSA-N Trp-Tyr-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)NCC(=O)O)N ZKVANNIVSDOQMG-HKUYNNGSSA-N 0.000 description 1
- XGEUYEOEZYFHRL-KKXDTOCCSA-N Tyr-Ala-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 XGEUYEOEZYFHRL-KKXDTOCCSA-N 0.000 description 1
- GFZQWWDXJVGEMW-ULQDDVLXSA-N Tyr-Arg-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N)O GFZQWWDXJVGEMW-ULQDDVLXSA-N 0.000 description 1
- CKKFTIQYURNSEI-IHRRRGAJSA-N Tyr-Asn-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 CKKFTIQYURNSEI-IHRRRGAJSA-N 0.000 description 1
- AYHSJESDFKREAR-KKUMJFAQSA-N Tyr-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AYHSJESDFKREAR-KKUMJFAQSA-N 0.000 description 1
- XMNDQSYABVWZRK-BZSNNMDCSA-N Tyr-Asn-Phe 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](CC1=CC=CC=C1)C(O)=O XMNDQSYABVWZRK-BZSNNMDCSA-N 0.000 description 1
- NXRGXTBPMOGFID-CFMVVWHZSA-N Tyr-Ile-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O NXRGXTBPMOGFID-CFMVVWHZSA-N 0.000 description 1
- FJBCEFPCVPHPPM-STECZYCISA-N Tyr-Ile-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O FJBCEFPCVPHPPM-STECZYCISA-N 0.000 description 1
- BYAKMYBZADCNMN-JYJNAYRXSA-N Tyr-Lys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O BYAKMYBZADCNMN-JYJNAYRXSA-N 0.000 description 1
- NVZVJIUDICCMHZ-BZSNNMDCSA-N Tyr-Phe-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O NVZVJIUDICCMHZ-BZSNNMDCSA-N 0.000 description 1
- SZEIFUXUTBBQFQ-STQMWFEESA-N Tyr-Pro-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SZEIFUXUTBBQFQ-STQMWFEESA-N 0.000 description 1
- BIWVVOHTKDLRMP-ULQDDVLXSA-N Tyr-Pro-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O BIWVVOHTKDLRMP-ULQDDVLXSA-N 0.000 description 1
- SYFHQHYTNCQCCN-MELADBBJSA-N Tyr-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O SYFHQHYTNCQCCN-MELADBBJSA-N 0.000 description 1
- MQUYPYFPHIPVHJ-MNSWYVGCSA-N Tyr-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC3=CC=C(C=C3)O)N)O MQUYPYFPHIPVHJ-MNSWYVGCSA-N 0.000 description 1
- AEOFMCAKYIQQFY-YDHLFZDLSA-N Tyr-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AEOFMCAKYIQQFY-YDHLFZDLSA-N 0.000 description 1
- RVGVIWNHABGIFH-IHRRRGAJSA-N Tyr-Val-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O RVGVIWNHABGIFH-IHRRRGAJSA-N 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- UEOOXDLMQZBPFR-ZKWXMUAHSA-N Val-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N UEOOXDLMQZBPFR-ZKWXMUAHSA-N 0.000 description 1
- YFOCMOVJBQDBCE-NRPADANISA-N Val-Ala-Glu Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N YFOCMOVJBQDBCE-NRPADANISA-N 0.000 description 1
- VMRFIKXKOFNMHW-GUBZILKMSA-N Val-Arg-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N VMRFIKXKOFNMHW-GUBZILKMSA-N 0.000 description 1
- OGNMURQZFMHFFD-NHCYSSNCSA-N Val-Asn-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N OGNMURQZFMHFFD-NHCYSSNCSA-N 0.000 description 1
- HZYOWMGWKKRMBZ-BYULHYEWSA-N Val-Asp-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HZYOWMGWKKRMBZ-BYULHYEWSA-N 0.000 description 1
- DLYOEFGPYTZVSP-AEJSXWLSSA-N Val-Cys-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)N1CCC[C@@H]1C(=O)O)N DLYOEFGPYTZVSP-AEJSXWLSSA-N 0.000 description 1
- AAOPYWQQBXHINJ-DZKIICNBSA-N Val-Gln-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N AAOPYWQQBXHINJ-DZKIICNBSA-N 0.000 description 1
- AHHJARQXFFGOKF-NRPADANISA-N Val-Glu-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N AHHJARQXFFGOKF-NRPADANISA-N 0.000 description 1
- VLDMQVZZWDOKQF-AUTRQRHGSA-N Val-Glu-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N VLDMQVZZWDOKQF-AUTRQRHGSA-N 0.000 description 1
- SYOMXKPPFZRELL-ONGXEEELSA-N Val-Gly-Lys Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)O)N SYOMXKPPFZRELL-ONGXEEELSA-N 0.000 description 1
- GQMNEJMFMCJJTD-NHCYSSNCSA-N Val-Pro-Gln Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O GQMNEJMFMCJJTD-NHCYSSNCSA-N 0.000 description 1
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 1
- PGQUDQYHWICSAB-NAKRPEOUSA-N Val-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N PGQUDQYHWICSAB-NAKRPEOUSA-N 0.000 description 1
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-GUBZILKMSA-N 0.000 description 1
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 1
- OFTXTCGQJXTNQS-XGEHTFHBSA-N Val-Thr-Ser Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](C(C)C)N)O OFTXTCGQJXTNQS-XGEHTFHBSA-N 0.000 description 1
- JVGDAEKKZKKZFO-RCWTZXSCSA-N Val-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)N)O JVGDAEKKZKKZFO-RCWTZXSCSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 125000000266 alpha-aminoacyl group Chemical group 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 238000012801 analytical assay Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000000628 antibody-producing cell Anatomy 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010068380 arginylarginine Proteins 0.000 description 1
- 108010060035 arginylproline Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 150000001510 aspartic acids Chemical class 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 238000000749 co-immunoprecipitation Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000009260 cross reactivity Effects 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- 108010069495 cysteinyltyrosine Proteins 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 108010085059 glutamyl-arginyl-proline Proteins 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
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 108010090037 glycyl-alanyl-isoleucine Proteins 0.000 description 1
- 108010019832 glycyl-asparaginyl-glycine Proteins 0.000 description 1
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 1
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 1
- 108010048994 glycyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 108010050343 histidyl-alanyl-glutamine Proteins 0.000 description 1
- 150000002460 imidazoles Chemical class 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229960003971 influenza vaccine Drugs 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010027338 isoleucylcysteine Proteins 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 230000028744 lysogeny Effects 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 229940043515 other immunoglobulins in atc Drugs 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 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
- 239000002244 precipitate Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 108010070643 prolylglutamic acid Proteins 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 238000001799 protein solubilization Methods 0.000 description 1
- 230000007925 protein solubilization Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000001226 reprecipitation Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000003345 scintillation counting Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 108010084932 tryptophyl-proline Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000000007 visual effect Effects 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
-
- 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
- C07K16/1018—Orthomyxoviridae, e.g. influenza virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- 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
- G01N2333/11—Orthomyxoviridae, e.g. influenza virus
-
- 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
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Pulmonology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Microbiology (AREA)
- Communicable Diseases (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
Abstract
본 발명은 항체 생산 포유동물 유래 융합파트너; 및 목적 항원;으로 이루어진 재조합 항원을 이용하여 목적 항원 특이적인 항체를 제조하는 방법, 상기 방법으로 제조된 항체, 상기 항체를 포함하는 바이러스 감염 진단용 조성물 및 바이러스 검출방법에 관한 것이다.
본 발명에 의하면, 재조합 항원을 숙주세포에서 올바르게 접힌 형태로 수용성 발현시킬 수 있어 생산된 항체의 에피토프 인식 능력 및 항체의 결합력을 높일 수 있고, 융합파트너에 대한 교차면역을 최소화시킴으로써 목적 항원 특이적인 항체를 생산할 수 있으므로, 본 발명의 목적 항원 특이적인 항체는 진단 또는 치료 용도뿐 아니라 생명공학 연구를 비롯한 다양한 의학적 목적으로 유용하게 사용될 수 있다.The present invention relates to an antibody-producing mammal-derived fusion partner; and a target antigen; to a method for producing a target antigen-specific antibody using a recombinant antigen consisting of, the antibody prepared by the above method, a composition for diagnosing a viral infection comprising the antibody, and a virus detection method.
According to the present invention, it is possible to soluble expression of a recombinant antigen in a properly folded form in a host cell, thereby increasing the epitope recognition ability of the produced antibody and the binding force of the antibody, and by minimizing cross-immunity to the fusion partner, the target antigen-specific antibody can be produced, the antigen-specific antibody of the present invention can be usefully used for various medical purposes, including diagnostic or therapeutic uses, as well as biotechnology research.
Description
본 발명은 항체 생산 숙주동물 유래 융합파트너; 및 목적 항원으로 이루어진 재조합 단백질을 이용하여 목적 항원 특이적인 항체를 제조하는 방법, 상기 방법으로 제조된 항체, 상기 항체를 포함하는 바이러스 감염 진단용 조성물 및 바이러스 검출방법에 관한 것이다.The present invention relates to a fusion partner derived from an antibody-producing host animal; And it relates to a method for producing a target antigen-specific antibody using a recombinant protein consisting of a target antigen, the antibody prepared by the above method, a composition for diagnosing a viral infection comprising the antibody, and a virus detection method.
항체는 현대 생명공학연구에 널리 사용되는 물질로 그 응용범위는 연구목적을 넘어 질병의 진단, 예방, 치료 등의 목적으로 광범위하게 사용되고 있다. 이러한 항체를 생산하는 가장 널리 알려진 방법은 항체 제작을 목적으로 하는 단백질 항원을 생산 숙주에 접종하여 숙주 내 면역시스템을 통해 항체를 생산하고 이 중 목적항원 특이적인 항체만을 스크리닝 하여 정제해 내는 방법이다. 따라서 접종하는 목적항원의 품질이 최종 생산품인 항체의 품질에도 커다란 영향을 주게 된다.Antibodies are widely used materials in modern biotechnology research, and their range of application goes beyond research purposes and is widely used for the purposes of diagnosis, prevention, and treatment of diseases. The most widely known method for producing such an antibody is a method of inoculating a production host with a protein antigen for the purpose of antibody production, producing the antibody through the immune system in the host, and screening and purifying only the target antigen-specific antibody among them. Therefore, the quality of the target antigen to be inoculated greatly affects the quality of the final product, the antibody.
단백질 항원의 품질을 결정하는 요소에는 크게 물속에서의 용해도(Solubility)와 올바른 구조의 형성여부(folding, 단백질 접힘)가 포함된다. 생명체 현상은 물속에서 진행되며 따라서 항원 단백질은 우선적으로 물에서 용해도가 높은 수용성 유지가 중요하다. 항원이 불수용성으로 발현된다는 것은 일반적으로 항원이 올바른 3차원 구조를 이루지 못하고 있음을 뜻하기 때문이다. 항원이 불용성일 경우 특히 항원의 3차원 구조에서 중요한 conformational epitope의 형성에 심각한 영향을 끼친다. 이를 해결하기 위해 일반적으로 불용성 침전을 리폴딩(refolding)이라는 단백질 수용화 추가 공정을 거치게 된다. 이에는 막대한 생산 비용 증가와 더불어 목표항원의 최종 수율(yield) 감소 등 산업상 단점이 제기되고 있다(7). 이에 더하여 리폴딩 항원은 원래의 자연적 구조로의 전환 미비(8-10) 등 항원의 품질이 저하될 우려가 있으며 이를 사용하여 항체를 제작할 경우 항체의 결합력, conformational epitope 인식능력 저하 등 항체의 질에 문제를 야기할 수 있다.Factors that determine the quality of a protein antigen include solubility in water and the formation of a correct structure (folding, protein folding). The biological phenomenon proceeds in water, and therefore, it is important to maintain water solubility with high solubility in water for antigenic proteins. This is because the expression of an antigen in a water-insoluble manner generally means that the antigen does not form a correct three-dimensional structure. When an antigen is insoluble, it seriously affects the formation of an important conformational epitope, especially in the three-dimensional structure of the antigen. In order to solve this problem, the insoluble precipitation is generally subjected to an additional process of protein solubilization called refolding. In addition to a huge increase in production cost, industrial disadvantages such as a decrease in the final yield of the target antigen are raised (7). In addition, refolding antigens may deteriorate the quality of the antigen, such as insufficient conversion to the original natural structure (8-10). It can cause problems.
이러한 재조합 단백질항원의 불수용성 침전형성 문제와 구조 접힘 문제해결을 위한 일환으로 목표항원을 수용성이 높은 단백질(Soluble carrier)에 융합하여 발현시키는 방법이 개발되었다(12). 그러나 이러한 융합단백질을 항원으로 면역하여 항체 생산 시에 융합파트너 자체가 면역동물에 외래 단백질로 인식되어 융합파트너에 대한 항체가 크게 유도되는 문제가 생기게 된다. 경우에 따라서는 융합파트너에 대한 면역원성이 목적항원을 압도하게 되며 불필요한 융합파트너에 대한 항체가 더 많이 유도될 수 있다. 즉 목표항원 특이 (specific) 항체 외에 융합파트너에 교차반응(cross-reactive)이 높은 항원이 혼재하게 된다. 이러한 문제점을 피하기 위해 유전자재조합 방법으로 얻은 융합단백질을 프로테아제(protease) 등으로 처리하여 융합파트너을 제거할 수 있다(19, 20). 이 경우 고가의 단백질절단효소가 필요할 뿐 아니라 추가적인 정제공정이 요구되고, 융합파트너 제거 이후 목적항원의 수용성 및 구조가 유지되지 않아 재침전으로 형성되는 문제가 있다.As a part of solving the problem of water-insoluble precipitate formation and structural folding of the recombinant protein antigen, a method for expressing a target antigen by fusing it to a highly water-soluble protein (Soluble carrier) was developed (12). However, when immunizing such a fusion protein with an antigen and producing an antibody, the fusion partner itself is recognized as a foreign protein by the immunized animal, resulting in a problem in that the antibody against the fusion partner is largely induced. In some cases, the immunogenicity to the fusion partner overwhelms the target antigen, and more antibodies to the unnecessary fusion partner may be induced. That is, in addition to the target antigen-specific antibody, antigens with high cross-reactivity are mixed with the fusion partner. In order to avoid this problem, the fusion protein obtained by the gene recombination method can be treated with a protease or the like to remove the fusion partner (19, 20). In this case, expensive proteolytic enzymes are required, an additional purification process is required, and the water solubility and structure of the target antigen are not maintained after removal of the fusion partner, so there is a problem of re-precipitation.
상기 종래기술의 문제점을 해결하기 위하여, 본 발명은 목적 항원의 올바른 발현을 유도하는 융합파트너를 포함하여 재조합 항원을 수용성으로 생성하고 이를 적절한 숙주 동물에 투여하여 목적 항원의 에피토프만 특이적으로 인식하는 항체를 제조하는 방법을 제공하는 데 있다.In order to solve the problems of the prior art, the present invention provides a method for specifically recognizing only an epitope of a target antigen by producing a recombinant antigen in a water-soluble manner, including a fusion partner that induces correct expression of the target antigen, and administering it to an appropriate host animal. To provide a method for producing an antibody.
본 발명의 다른 목적은 상기 방법으로 제조된 목적 항원 특이적 항체, 이를 포함하는 바이러스 진단용 조성물 및 이를 이용하여 바이러스를 검출하는 방법을 제공하는 데 있다.Another object of the present invention is to provide a target antigen-specific antibody prepared by the above method, a composition for virus diagnosis comprising the same, and a method for detecting a virus using the same.
상기 목적을 달성하기 위하여 본 발명은 목적 항원의 발현 효율을 증진시키기 위한 융합파트너; 및 목적 항원으로 이루어지는 재조합 단백질을 항체 생산 숙주동물에 투여하는 단계를 포함하는 항체 제조방법을 제공한다.In order to achieve the above object, the present invention provides a fusion partner for enhancing the expression efficiency of a target antigen; and administering a recombinant protein composed of a target antigen to an antibody-producing host animal.
본 발명은 또한, 상기 제조방법에 의해 제조된, 서열번호 4, 서열번호 5 및 서열번호 6으로 이루어진 군에서 선택된 1종 이상의 펩타이드를 특이적으로 인지하는 목적 항원 특이적 항체를 제공한다.The present invention also provides a target antigen-specific antibody specifically recognizing one or more peptides selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, prepared by the above production method.
본 발명은 또한, 서열번호 4, 서열번호 5 및 서열번호 6으로 이루어진 군에서 선택된 1종 이상의 아미노산 서열로 이루어진 펩타이드를 특이적으로 인지하는 목적 항원 특이적 항체를 포함하는 바이러스 감염 진단용 조성물을 제공한다.The present invention also provides a composition for diagnosing viral infection comprising a target antigen-specific antibody that specifically recognizes a peptide consisting of one or more amino acid sequences selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6. .
본 발명은 또한, 샘플과 항체를 접촉시키는 단계; 및 상기 항체와 샘플의 반응을 검출하는 단계를 포함하는, 대상체의 바이러스 감염 여부에 대한 정보제공 방법을 제공한다.The present invention also provides a method comprising: contacting a sample with an antibody; And it provides a method of providing information on whether a subject is infected with a virus, comprising the step of detecting a reaction between the antibody and the sample.
본 발명의 항체 제조방법에 의하면, 융합파트너를 포함하여 재조합 항원을 숙주세포에서 올바르게 접힌 형태로 수용성 발현시킴으로써 항체의 에피토프 인식 능력 및 항체의 결합력을 높일 수 있다.According to the antibody production method of the present invention, the ability of the antibody to recognize the epitope and the binding ability of the antibody can be increased by soluble expression of the recombinant antigen including the fusion partner in a properly folded form in the host cell.
또한, 포유동물 유래 융합파트너를 사용하여 생성된 재조합 항원을 동일한 포유 동물에 투여하여 융합파트너에 대한 교차면역을 최소화함으로써 목적 항원 특이적인 항체를 생산할 수 있으며, 본 발명의 RID 융합파트너는 다양한 포유동물에 존재하는 단백질로 다양한 항체 생산 숙주동물에 알맞은 재조합 항원 생산에 사용될 수 있는 장점이 있다.In addition, by administering a recombinant antigen generated using a mammal-derived fusion partner to the same mammal, cross-immunity against the fusion partner can be minimized to produce an antigen-specific antibody of interest, and the RID fusion partner of the present invention can be used in a variety of mammals. It has the advantage that it can be used for the production of recombinant antigens suitable for various antibody-producing host animals as a protein present in
본 발명의 목적 항원 특이적인 항체는 진단 또는 치료 용도뿐 아니라 생명공학 연구를 비롯한 다양한 의학적 목적으로 유용하게 사용될 수 있다.The antigen-specific antibody of the present invention can be usefully used for various medical purposes, including not only diagnostic or therapeutic uses, but also biotechnology research.
도 1은 RID 융합을 통한 항원 및 항체 제작 방법에 대한 개략적 도면이다.
도 1A는 쥐 유래 RID를 융합단백질로 이용하여 목적단백질에 대한 쥐 유래 단일클론항체 및 다클론항체 생산과정의 개략적 도식이다.
도 1B는 Lysyl tRNA synthetase 중 RNA interaction domain(RID)의 위치에 대한 개략 도식과 더불어 쥐, 토끼, 인간 유래 RID의 아미노산 서열이다(24).
도 2는 본 발명을 통해 인플루엔자 재조합 헤마글루티닌(Hemagglutinin, HA) 단백질의 stalk 도메인을 대장균 발현 시스템에서의 발현 결과에 대한 도면이다. 발현하고자 하는 목적 단백질은 인플루엔자 A 바이러스 그룹 1에 해당하는 여러 종의 인플루엔자 바이러스의 HA 단백질 서열 중 공통 아미노산 서열을 포함하도록 디자인한 유니버설 HA 단백질이다.
도 2A는 유니버설 HA 단백질 발현을 위한 mRID 융합 벡터, mRID 융합 유니버설 HA 단백질(mRID-uHAstalk), mRID를 융합하지 않은 유니버설 HA 단백질(uHA stalk)의 모식도이다.
도 2B는 mRID-uHAstalk 37°C 와 20°C 에서의 발현 양상을 보여주는 도면이다. M은 단백질 크기 마커, T는 전체 단백질 분획, S는 수용성 단백질 분획, P는 불수용성 단백질 분획을 나타낸다. IPTG(Isopropyl β- D -1-thiogalactopyranoside)를 첨가 여부는 (-) 와 (+) 로 표기하였다. 도 2C는 mRID를 융합하지 않은 유니버설 HA 단백질(uHA stalk)의 37°C 와 20°C 에서의 발현 양상을 보여주는 SDS-PAGE(SDS-polyacrylamide gel electrophoresis) 결과이다.
도 3은 mRID-uHAstalk을 접종하여 쥐에서 생산된 하이브리도마 클론과 이중 정제된 단일클론항체 2종(1G5, 2C12)을 이용하여 융합단백질 mRID와 접종에 사용된 mRID-uHAstalk 대하여 ELISA을 진행한 결과이다.
도 3A는 항체가 포함된 하이브리도마 클론 15종을 이용한 ELSIA 결과이다.
도 3B는 1G5 항체를 이용한 ELISA 결과이며, 도 3C는 2C12 항체를 이용한 ELISA 결과이다.
도 4는 유니버설 HA 단백질에 대한 단일클론항체 1G5를 이용하여 실제 인플루엔자 바이러스의 HA 단백질에 대해 ELISA를 진행한 결과이다. 위 ELISA에서 사용된 인플루엔자라 바이러스 HA는 영국 NIBSC(National Institute for Biological Standards and Control)에서 공급된 표준 HA 단백질 항원이다.
도 5는 유니버설 HA 단백질에 대한 단일클론항체 2C12를 이용하여 실제 인플루엔자 바이러스의 HA 단백질에 대해 ELISA를 진행한 결과이다. 위 ELISA에서 사용된 인플루엔자라 바이러스 HA는 영국 NIBSC(National Institute for Biological Standards and Control)에서 공급된 표준 HA단백질 항원이다.
도 6은 인플루엔자 A/Puerto Rico/08/34(H1N1) 바이러스의 HA 단백질의 globular head domain(HAgd)을 대장균 발현 시스템에서의 발현 결과에 대한 도면이다.
도 6A는 HAgd 단백질의 발현을 위해 사용된 플라스미드 벡터와 RID 융합 HAgd 단백질(RID-HAgd)의 모식도이다.
도 6B는 RID-HAgd 과 RID를 융합하지 않은 HAgd의 20°C에서 발현 양상을 보여주는 SDS-PAGE 결과이다. M은 단백질 크기 마커, T는 전체 단백질 분획, S는 수용성 단백질 분획, P는 불수용성 단백질 분획을 나타낸다. IPTG(Isopropyl β- D -1-thiogalactopyranoside )를 첨가 여부는 (-) 와 (+) 로 표기하였다.
도 7은 mRID-HAgd 와 rRID-HAgd 단백질의 접힘 적합성, 위 단백질을 통해 생산된 항혈청 내 항체의 반응성을 보여주는 도면이다.
도 7A는 야생형 인플루엔자 A/Puerto Rico/08/34(H1N1) 바이러스를 쥐에 접종 후 얻은 항혈청을 이용해 mRID-HAgd, mRID, HAgd에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 7B는 야생형 인플루엔자 A/Puerto Rico/08/34(H1N1) 바이러스를 쥐에 접종 후 얻은 항혈청을 이용해 rRID-HAgd, rRID, HAgd에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 7C는 여러 종류의 인플루엔자 바이러스를 접종하여 얻은 쥐 항혈청을 이용해 mRID-HAgd에 대하여 ELISA를 수행한 결과이다. A/Puerto Rico/08/34 (H1N1), X-31 기반 저온적응 A/New Caledonia/20/99 (H1N1), X-31 기반 저온적응 A/Panama/07/99 (H3N2), X-31 기반 저온적응 A/Indonesia/5/2005 (H5N1), X-31 기반 저온적응 A/Korea/01/09 (pandemic H1N1) 를 쥐에 접종하여 얻은 항혈청(26, 27)이 사용되었다.
도 7D는 mRID-HAgd를 쥐에 접종한 후 얻은 항혈청을 이용해 mRID-HAgd, mRID, HAgd에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 7E는 rRID-HAgd를 토끼에 접종한 후 얻은 항혈청을 이용해 rRID-HAgd, rRID, HAgd에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 7F는 대장균 유래 Lysyl tRNA synthetase (LysRS) 를 융합한 HAgd(LysRS-HAgd)를 쥐에 접종한 후 얻은 항혈청을 이용해 LysRS-HAgd, LysRS에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 8은 mRID 및 rRID 융합 EGFP(Enhanced Green Fluorescent Protein)의 발현과 이에 대한 쥐 항혈청 내 항체의 반응성을 보여주는 도면이다.
도 8A는 RID융합 EGFP (RID-EGFP) 발현을 위해 사용된 플라스미드 벡터와 RID-EGFP의 모식도이다. 사용된 융합단백질은 mRID와 rRID이다.
도 8B는 mRID 융합 EGFP(mRID-EGFP), rRID 융합 EGFP(rRID-EGFP), 융합하지 않은 EGFP 단백질의 25°C에서의 발현양상을 보여주는 SDS-PAGE 결과이다. M은 단백질 크기 마커, T는 전체 단백질 분획, S는 수용성 단백질 분획, P는 불수용성 단백질 분획을 나타낸다. IPTG(Isopropyl β- D -1-thiogalactopyranoside )를 첨가 여부는 (-) 와 (+) 로 표기하였다.
도 8C는 mRID-EGFP 단백질을 쥐에 접종하여 얻은 항혈청을 이용해 mRID-EGFP, mRID, EGFP에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 8D는 rRID-EGFP 단백질을 토끼에 접종하여 얻은 항혈청을 이용해 rRID-EGFP, rRID, EGFP에 대하여 ELISA와 Western blot을 수행한 결과이다.
도 9는 mRID 융합 MERS(Middle East respiratory syndrome coronavirus)의 S1 항원의 RBD(Receptor Binding Domain) 발현과 이에 대한 단일클론항체의 반응성을 보여주는 도면이다.
도 9A는 mRID융합 MERS RBD(mRID-MERS RBD) 발현을 위해 사용된 플라스미드 벡터와 mRID-MERS RBD의 모식도이다.
도 9B는 mRID를 융합하지 않은 MERS-RBD의 16°C에서의 발현 양상을 보여주는 SDS-PAGE 결과이다. M은 단백질 크기 마커, T는 전체 단백질 분획, S는 수용성 단백질 분획, P는 불수용성 단백질 분획을 나타낸다. IPTG(Isopropyl - D -1-thiogalactopyranoside )를 첨가 여부는 (-) 와 (+) 로 표기하였다.
도 9C는 mRID-MERS RBD)의 37°C, 30°C, 20°C, 16°C 에서의 발현 양상을 보여주는 SDS-PAGE 결과이다. M은 단백질 크기 마커, T는 전체 단백질 분획, S는 수용성 단백질 분획, P는 불수용성 단백질 분획을 나타낸다. IPTG(Isopropyl β- D -1-thiogalactopyranoside )를 첨가 여부는 (-) 와 (+) 로 표기하였다.
도 9D는 293T 세포에서 생산된 RBD단백질, mRID-MERS RBD 단백질, mRID 단백질과 hDPP4(human dipeptidyl peptidase4) 단백질 사이의 결합 여부와 정도를 ELISA를 통해 분석한 결과이다.
도 9E는 mRID-MERS RBD단백질을 쥐에 접종하여 생산한 단일클론항체 2종(mAb29, mAb38)을 이용하여 Baculovirus-insect cell에서 생산된 RBD 단백질항원에 대한 ELISA를 진행한 결과이다.
도 9F는 mRID-MERS RBD 단백질에 대한 단일클론항체 mAb29를 이용하여 mRID-MERS RBD, mRID, 대장균 수용성 세포(competent cell) 내재 단백질에 대한 Western blot을 진행한 결과이다.1 is a schematic diagram of an antigen and antibody production method through RID fusion.
1A is a schematic diagram of the production process of a mouse-derived monoclonal antibody and a polyclonal antibody against a target protein using a mouse-derived RID as a fusion protein.
1B is a schematic diagram of the position of the RNA interaction domain (RID) in Lysyl tRNA synthetase, along with the amino acid sequence of RID derived from rat, rabbit, and human (24).
2 is a view showing the expression results of the stalk domain of influenza recombinant hemagglutinin (HA) protein in the E. coli expression system through the present invention. The target protein to be expressed is a universal HA protein designed to include a common amino acid sequence among the HA protein sequences of several influenza viruses corresponding to influenza
2A is a schematic diagram of an mRID fusion vector for universal HA protein expression, mRID fusion universal HA protein (mRID-uHAstalk), and universal HA protein (uHA stalk) without mRID fusion.
Figure 2B is a diagram showing the expression pattern of mRID-uHAstalk at 37 °C and 20 °C. M denotes a protein size marker, T denotes the total protein fraction, S denotes a soluble protein fraction, and P denotes a water-insoluble protein fraction. Whether or not IPTG (Isopropyl β-D -1-thiogalactopyranoside) was added was indicated by (-) and (+). Figure 2C is the result of SDS-PAGE (SDS-polyacrylamide gel electrophoresis) showing the expression pattern at 37 °C and 20 °C of the universal HA protein (uHA stalk) not fused with mRID.
Figure 3 shows the ELISA for the fusion protein mRID and mRID-uHAstalk used for inoculation using a hybridoma clone produced in mice by inoculation with mRID-uHAstalk and two types of double purified monoclonal antibodies (1G5, 2C12). is the result
3A is an ELSIA result using 15 hybridoma clones containing antibodies.
Fig. 3B is an ELISA result using the 1G5 antibody, and Fig. 3C is an ELISA result using the 2C12 antibody.
4 is a result of ELISA for the HA protein of the actual influenza virus using the monoclonal antibody 1G5 against the universal HA protein. Influenza virus HA used in the above ELISA is a standard HA protein antigen supplied by the British National Institute for Biological Standards and Control (NIBSC).
5 is a result of ELISA for the HA protein of the actual influenza virus using the monoclonal antibody 2C12 against the universal HA protein. Influenza virus HA used in the above ELISA is a standard HA protein antigen supplied by the National Institute for Biological Standards and Control (NIBSC) in the UK.
6 is a view showing the expression results of the globular head domain (HAgd) of the HA protein of influenza A / Puerto Rico / 08 / 34 (H1N1) virus in E. coli expression system.
6A is a schematic diagram of a plasmid vector and RID fusion HAgd protein (RID-HAgd) used for the expression of HAgd protein.
6B is an SDS-PAGE result showing the expression pattern of RID-HAgd and HAgd without RID fusion at 20 °C. M denotes a protein size marker, T denotes the total protein fraction, S denotes a soluble protein fraction, and P denotes a water-insoluble protein fraction. Whether or not IPTG (Isopropyl β-D -1-thiogalactopyranoside) was added was indicated by (-) and (+).
7 is a view showing the folding compatibility of mRID-HAgd and rRID-HAgd proteins, and the reactivity of antibodies in antisera produced through the above proteins.
7A shows the results of ELISA and Western blot for mRID-HAgd, mRID, and HAgd using antisera obtained after inoculation of wild-type influenza A/Puerto Rico/08/34 (H1N1) virus into mice.
7B shows the results of ELISA and Western blot for rRID-HAgd, rRID, and HAgd using antisera obtained after inoculation of wild-type influenza A/Puerto Rico/08/34 (H1N1) virus into mice.
7C is a result of performing ELISA for mRID-HAgd using mouse antiserum obtained by inoculating several types of influenza virus. A/Puerto Rico/08/34 (H1N1), X-31-based low-temperature adaptation A/New Caledonia/20/99 (H1N1), X-31-based low-temperature adaptation A/Panama/07/99 (H3N2), X-31 Antisera (26, 27) obtained by inoculating mice with cold-adapted A/Indonesia/5/2005 (H5N1) and X-31-based cold-adapted A/Korea/01/09 (pandemic H1N1) mice were used.
7D shows the results of ELISA and Western blot for mRID-HAgd, mRID, and HAgd using antisera obtained after inoculating mice with mRID-HAgd.
7E shows the results of ELISA and Western blot for rRID-HAgd, rRID, and HAgd using antisera obtained after inoculating rabbits with rRID-HAgd.
7F shows the results of ELISA and Western blot for LysRS-HAgd and LysRS using antiserum obtained after inoculating mice with HAgd (LysRS-HAgd) fused with E. coli-derived Lysyl tRNA synthetase (LysRS).
8 is a diagram showing the expression of mRID and rRID fusion Enhanced Green Fluorescent Protein (EGFP) and the reactivity of the antibody in rat antisera thereto.
8A is a schematic diagram of a plasmid vector and RID-EGFP used for RID fusion EGFP (RID-EGFP) expression. The fusion proteins used are mRID and rRID.
8B is an SDS-PAGE result showing the expression patterns of mRID fusion EGFP (mRID-EGFP), rRID fusion EGFP (rRID-EGFP), and unfused EGFP proteins at 25 °C. M denotes a protein size marker, T denotes the total protein fraction, S denotes a soluble protein fraction, and P denotes a water-insoluble protein fraction. Whether or not IPTG (Isopropyl β-D -1-thiogalactopyranoside) was added was indicated by (-) and (+).
FIG. 8C shows the results of ELISA and Western blot for mRID-EGFP, mRID, and EGFP using antisera obtained by inoculating mice with mRID-EGFP protein.
8D shows the results of ELISA and Western blot for rRID-EGFP, rRID, and EGFP using antisera obtained by inoculating rabbits with rRID-EGFP protein.
9 is a view showing the expression of receptor binding domain (RBD) of the S1 antigen of mRID fusion MERS (Middle East Respiratory Syndrome coronavirus) and the reactivity of the monoclonal antibody thereto.
9A is a schematic diagram of a plasmid vector and mRID-MERS RBD used for expression of mRID-fusion MERS RBD (mRID-MERS RBD).
9B is an SDS-PAGE result showing the expression pattern at 16 °C of MERS-RBD not fused with mRID. M denotes a protein size marker, T denotes the total protein fraction, S denotes a soluble protein fraction, and P denotes a water-insoluble protein fraction. Whether IPTG (Isopropyl -D -1-thiogalactopyranoside) was added was indicated by (-) and (+).
9C is an SDS-PAGE result showing the expression patterns of mRID-MERS RBD) at 37 °C, 30 °C, 20 °C, and 16 °C. M denotes a protein size marker, T denotes the total protein fraction, S denotes a soluble protein fraction, and P denotes a water-insoluble protein fraction. Whether or not IPTG (Isopropyl β-D -1-thiogalactopyranoside) was added was indicated by (-) and (+).
FIG. 9D shows the results of analyzing the binding between RBD protein, mRID-MERS RBD protein, mRID protein and hDPP4 (human dipeptidyl peptidase4) protein produced in 293T cells by ELISA.
9E is a result of ELISA for RBD protein antigen produced in Baculovirus-insect cells using two monoclonal antibodies (mAb29, mAb38) produced by inoculating mice with mRID-MERS RBD protein.
Figure 9F shows the results of Western blot for mRID-MERS RBD, mRID, and E. coli soluble cells (competent cell) intrinsic protein using the monoclonal antibody mAb29 against the mRID-MERS RBD protein.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명에서는 목적 항원의 발현 효율을 증진시키기 위한 융합파트너를 특정 길이 및 특정 서열로 포함하여 융합파트너-목적 항원의 재조합 단백질을 올바르게 접힌 형태로 수용성 발현시키고, 이를 포유동물에 투여하여 항체를 제조함에 있어서, 동일한 포유동물 유래의 융합파트너를 사용함으로써 생성된 항체가 목적 항원에만 특이적으로 결합하고, 융합파트너에는 결합하지 않는 것을 확인하였으며, 재조합 항원 단백질을 면역적으로 가치 있는 구조로 제시하여 진단용 또는 치료용 항체의 특이도를 높이는 것을 확인하고 본 발명을 완성하게 되었다.In the present invention, by including a fusion partner in a specific length and specific sequence to enhance the expression efficiency of the target antigen, the fusion partner-target antigen is soluble in a properly folded form to produce an antibody, and then administered to a mammal. In this case, it was confirmed that the antibody generated by using a fusion partner derived from the same mammal specifically binds only to the target antigen and does not bind to the fusion partner, and presents the recombinant antigen protein as an immunologically valuable structure for diagnostic or It was confirmed that the specificity of the therapeutic antibody was increased, and the present invention was completed.
이에, 본 발명은 목적 항원의 발현 효율을 증진시키기 위한 융합파트너; 및 목적 항원으로 이루어지는 재조합 단백질을 포유동물에 투여하는 단계를 포함하는 항체 제조방법에 있어서, 상기 융합파트너는 포유동물로부터 유래된 아미노아실 tRNA 합성효소에서 분리한 RNA 결합 도메인(Aminoacyl tRNA synthetase appended RNA interacting domain; RID)의 아미노산 서열을 포함하는 것인, 항체 제조방법을 제공한다.Accordingly, the present invention provides a fusion partner for enhancing the expression efficiency of a target antigen; and administering a recombinant protein comprising a target antigen to a mammal, wherein the fusion partner is an RNA-binding domain (Aminoacyl tRNA synthetase appended RNA interacting) isolated from an aminoacyl tRNA synthetase derived from a mammal. It provides a method for producing an antibody, including the amino acid sequence of the domain; RID).
하기 [도 1]은 마우스를 생산 숙주로 하여 마우스에서 유래된 RID를 융합 단백질로 이용하여 재조합 항원을 생산하는 과정 및 재조합 항원에 대한 단일클론항체 및 다클론항체를 제작하는 방법을 개략적으로 보여주는 개념도이다.The following [FIG. 1] is a conceptual diagram schematically showing a process for producing a recombinant antigen using a mouse as a production host and a mouse-derived RID as a fusion protein and a method for preparing monoclonal and polyclonal antibodies against the recombinant antigen. to be.
본 발명의 “목적 항원(target antigen)”은 당업자가 항체를 통해 검출하고자 하는 단백질로서, 재조합 발현벡터에 상기 단백질을 암호화하는 폴리뉴클레오티드를 삽입하여 숙주세포에서 발현이 가능한 모든 단백질을 의미한다. 본 발명에서 “목적 항원”은 숙주세포에서 융합파트너과 융합된 형태로 발현된다. 본 발명의 실시예에서는 “목적 항원”으로 인플루엔자바이러스 유래 uHAstalk(서열번호 4의 아미노산 서열로 표시됨)와 HAgd(서열번호 5의 아미노산 서열로 표시됨) 및 메르스 코로나 바이러스 유래 S1 단백질의 RBD(서열번호 6의 아미노산 서열로 표시됨)를 사용하였다.The "target antigen" of the present invention refers to any protein that can be expressed in a host cell by inserting a polynucleotide encoding the protein into a recombinant expression vector as a protein that those skilled in the art want to detect through an antibody. In the present invention, the "target antigen" is expressed in a form fused with a fusion partner in a host cell. In an embodiment of the present invention, as “target antigens”, influenza virus-derived uHAstalk (represented by the amino acid sequence of SEQ ID NO: 4) and HAgd (represented by the amino acid sequence of SEQ ID NO: 5) and MERS coronavirus-derived S1 protein RBD (SEQ ID NO: 6) was used.
본 발명의 "목적 단백질의 발현 효율을 증진시키기 위한 융합파트너"는 목적 단백질을 수용성 형태로 발현시킬 수 있다고 보고된 펩타이드로서, 예를 들어, 잘 알려진 융합파트너로서 GST, 말토스 결합 단백질, 유비퀴틴 등이 있으나 이들은 일반적인 포유동물에 접종 시 외래항원으로 인식되어 이에 대한 항체 유도가 발생된다. 본 발명에서 사용하고자 하는 융합 파트너는 일반적인 융합파트너와는 다르게 생산숙주 유래 단백질을 Aminoacyl tRNA 합성요소의 RNA interaction domain, 특히 라이실 tRNA 합성요소의 RNA interaction domain (RBD) 를 사용함으로써 융합파트너에 대한 면역유도를 최소화하며 목적항원에 대한 항체 생산을 최대화 할 수 있다.The "fusion partner for enhancing the expression efficiency of the target protein" of the present invention is a peptide reported to be capable of expressing the target protein in a water-soluble form, for example, as well-known fusion partners such as GST, maltose binding protein, ubiquitin, etc. However, they are recognized as foreign antigens when inoculated into general mammals, and antibody induction occurs. Unlike general fusion partners, the fusion partner to be used in the present invention is immune to the fusion partner by using the RNA interaction domain of the aminoacyl tRNA synthesizing element, particularly the RNA interaction domain (RBD) of the lysyl tRNA synthesizing element, for the protein derived from the production host. Induction can be minimized and antibody production against the target antigen can be maximized.
본 명세서에서 사용된 용어 “RID(RNA interacting domain)”은 RNA와 기타 단백질 간의 상호작용에 관여하는 고유한 N-말단 연장부위(70 아미노산)를 의미하며, 라이실 tRNA 합성 효소로부터 분리한 것이다. 본 발명의 바람직한 일 실시예에서는, RID에 태그 서열을 결합한 tag-RID를 박테리아에서 목적 단백질의 수용성 발현을 위한 결합 파트너로 사용하였다.As used herein, the term “RNA interacting domain (RID)” refers to a unique N-terminal extension (70 amino acids) involved in the interaction between RNA and other proteins, and is isolated from lysyl tRNA synthetase. In a preferred embodiment of the present invention, tag-RID in which a tag sequence is bound to RID was used as a binding partner for soluble expression of a target protein in bacteria.
본 발명의 바람직한 일실시예에서, 상기 RID는 RID-목적 항원을 도입하여 항체를 생산하기 위한 포유동물로부터 유래된 아미노아실 tRNA 합성효소에서 분리된 RNA 결합 도메인일 수 있고, 예를 들어 서열번호 1로 표시되는 마우스 유래 RID, 서열번호 2로 표시되는 토끼 유래 RID, 서열번호 3으로 표시되는 인간 유래 RID일 수 있다.In a preferred embodiment of the present invention, the RID may be an RNA binding domain isolated from an aminoacyl tRNA synthetase derived from a mammal for introducing a RID-target antigen to produce an antibody, for example, SEQ ID NO: 1 It may be a mouse-derived RID represented by , a rabbit-derived RID represented by SEQ ID NO: 2, or a human-derived RID represented by SEQ ID NO: 3.
본 발명의 항체의 제조방법에 있어서, 상기 언급된 융합단백질에 대한 항체 유도 문제를 해결하기 위해 항체 생산 숙주 동물로부터 유래한 자가항원(self-antigen)을 융합파트너로 사용할 수 있다. 이 경우 융합파트너가 자가항원('self ' antigen)으로 인식되어 이에 대한 면역반응이 일어나지 않게 되고, 면역동물에는 이종항원('foreign' antigen)인 목적 항원에 대해서만 면역이 유도되게 된다. 이에, 면역 숙주 동물 유래 단백질을 융합파트너로 이용해 항원 단백질의 수용성 향상과 올바른 구조로의 접힘을 도모함과 동시에 융합파트너에 대한 면역 유도를 방지하여 목적항원에 대한 특이적인 항체 생산이 가능하도록 할 수 있다.In the method for producing an antibody of the present invention, a self-antigen derived from an antibody-producing host animal may be used as a fusion partner in order to solve the above-mentioned problem of inducing the antibody to the fusion protein. In this case, the fusion partner is recognized as a 'self' antigen, and an immune response to it does not occur, and immunity is induced only to the target antigen, which is a 'foreign' antigen, in the immunized animal. Accordingly, by using an immune host animal-derived protein as a fusion partner, it is possible to improve the solubility of the antigen protein and to fold it into the correct structure, and at the same time prevent the induction of immunity to the fusion partner, thereby enabling the production of antibodies specific to the target antigen. .
다른 측면에서, 본 발명은 서열번호 4, 서열번호 5 및 서열번호 6으로 이루어진 군에서 선택된 1종 이상의 펩타이드를 특이적으로 인지하는 목적 항원 특이적 항체를 제공한다.In another aspect, the present invention provides a target antigen-specific antibody specifically recognizing one or more peptides selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.
상기 항체는 단일클론 항체 또는 다클론 항체 일 수 있다.The antibody may be a monoclonal antibody or a polyclonal antibody.
본 발명의 일 구현예에 의하면, 인플루엔자바이러스 유래 uHAstalk(서열번호 4의 아미노산 서열로 표시됨)와 HAgd(서열번호 5의 아미노산 서열로 표시됨) 및 메르스 코로나 바이러스 유래 S1 항원의 RBD(서열번호 6의 아미노산 서열로 표시됨)를 코딩하는 핵산을 벡터에 삽입하고, 이 벡터를 특정 대장균 균주로 접종하여 형질전환시킴으로써, 상기 단백질이 과도하게 발현된 균주로부터 수득한 재조합 단백질을 사용하여 생산 숙주 동물을 면역화하고 이로부터 적출한 비장세포와 골수종 세포를 융합하여 얻어진 하이브리도마 중에서 다양한 바이러스로부터 발현되는 상기 단백질을 항원으로 인식하여 이에 대해서만 특이적으로 반응하는 하이브리도마 세포주를 선별하였다.According to one embodiment of the present invention, influenza virus-derived uHAstalk (represented by the amino acid sequence of SEQ ID NO: 4) and HAgd (represented by the amino acid sequence of SEQ ID NO: 5) and RBD of the MERS coronavirus-derived S1 antigen (SEQ ID NO: 6) immunization of a production host animal with a recombinant protein obtained from a strain in which the protein is overexpressed by inserting a nucleic acid encoding the amino acid sequence) into a vector, inoculating the vector with a specific E. coli strain, and transforming the vector; Among the hybridomas obtained by fusion of splenocytes and myeloma cells extracted therefrom, hybridoma cell lines that recognize the proteins expressed from various viruses as antigens and respond specifically to them were selected.
본 발명은 대다수의 항체 생산에 일반적으로 사용하는 마우스 면역을 중심으로 기술되었다. 그러나 이는 용이하게 마우스 이외의 숙주 동물, 예를 들어 토끼, 말, 개, 고양이와 같은 포유류 동물, 닭, 오리 등과 같은 조류 등 다양한 숙주 동물로 확대 적용될 수 있다. 예를 들어 토끼를 생산 숙주로 하여 재조합 항원에 대한 토끼 유래 항체 제작 시 목적 항원에 토끼 유래 RID를 융합, 사용함으로서 목적항원 특이적 항체 생산의 효율성을 증가시킬 수 있다.The present invention has been described with a focus on mouse immunity commonly used for the production of the majority of antibodies. However, it can be easily applied to host animals other than mice, for example, mammals such as rabbits, horses, dogs, and cats, and various host animals such as birds such as chickens and ducks. For example, when producing a rabbit-derived antibody against a recombinant antigen using a rabbit as a production host, the efficiency of production of the target antigen-specific antibody can be increased by fusing and using the rabbit-derived RID to the target antigen.
본 발명의 단일클론항체는 본 기술분야에서 잘 알려져 있는 융합 방법에 의하여 제작될 수 있다. 주지한 것처럼 단일클론항체를 분비하는 하이브리도마 세포는 항원을 주사하여 면역시킨 마우스의 면역 세포와 적절한 암 세포주를 융합하는 방법으로 제작할 수 있다. 마우스의 면역 세포와 암 세포주의 세포 융합은 폴리에틸렌글리콜(PEG)와 같이 잘 알려진 방법을 이용할 수 있으며, 항체 생산 세포 역시 일반적인 배양 방법으로 증식시킨다.The monoclonal antibody of the present invention can be prepared by a fusion method well known in the art. As is well known, hybridoma cells secreting monoclonal antibodies can be prepared by fusing immune cells from mice immunized with antigen and an appropriate cancer cell line. Cell fusion of mouse immune cells and cancer cell lines can use a well-known method such as polyethylene glycol (PEG), and antibody-producing cells are also proliferated by a general culture method.
본 명세서에서 사용된 용어 '단일클론 항체'란 당해 분야에 공지된 용어로서 단일 항원성 부위에 대해서 지시되는 고도의 특이적인 항체를 의미한다. 단일클론 항체는 단세포군 항체라고 불리기도 한다. 통상적으로, 상이한 에피토프(항원결정기)들에 대해 지시되는 상이한 항체들을 포함하는 다클론 항체와는 다르게, 단일클론항체는 항원상의 단일 결정기에 대해서 지시된다. 단일클론 항체는 항원-항체 결합을 이용하는 진단 및 분석학적 분석법의 선택성과 특이성을 개선시키는 장점이 있으며, 또한 하이브리도마 배양에 의해 합성되기 때문에 다른 면역글로불린에 의해 오염되지 않는 또 다른 장점을 갖는다.As used herein, the term 'monoclonal antibody' is a term known in the art and refers to a highly specific antibody directed against a single antigenic site. Monoclonal antibodies are also called unicellular antibodies. Unlike polyclonal antibodies, which typically include different antibodies directed against different epitopes (epitopes), monoclonal antibodies are directed against a single determinant on an antigen. Monoclonal antibodies have the advantage of improving the selectivity and specificity of diagnostic and analytical assays using antigen-antibody binding, and also have another advantage of not being contaminated by other immunoglobulins because they are synthesized by hybridoma culture.
상기 하이브리도마가 생산하는 단일클론 항체는 정제하지 않고 사용할 수도 있으나, 최선의 결과를 얻기 위해서는 본 발명이 속하는 기술분야에 잘 알려져 있는 방법에 따라 고순도(예컨대, 95% 이상)로 정제하여 사용하는 것이 바람직하다. 이러한 정제 기술로는, 예를 들어 겔 전기영동, 투석, 염 침전, 이온교환 크로마토 그래피, 친화성 크로마토그래피 등의 정제 방법을 이용하여 배양 배지 또는 복수액으로부터 분리될 수 있다.The monoclonal antibody produced by the hybridoma may be used without purification, but in order to obtain the best results, it is purified with high purity (eg, 95% or more) according to a method well known in the art to which the present invention belongs. it is preferable As such a purification technique, for example, gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, affinity chromatography, etc. may be used to separate from the culture medium or ascites fluid.
본 발명의 일 구현예에서, 본 발명의 단일클론 항체는 인플루엔자 바이러스의 mRID-uHAstalk 항원을 사용하여 면역된 마우스 비장세포로부터 세포융합기술에 의해 획득한 하이브리도마 세포들로부터 생산되는 항체를 제작하였으며, 이를 1G5 및 2C12로 명명하였다. 본 발명의 또 다른 구현예에서, 본 발명의 단일클론 항체는 메르스 코로나 바이러스의 mRID-MERS RBD 항원을 사용하여 면역된 마우스 비장세포로부터 세포융합기술에 의해 획득한 하이브리도마 세포들로부터 생산되는 항체를 제작하였으며, 이를 각각 mAb29 및 mAb38로 명명하였다.In one embodiment of the present invention, the monoclonal antibody of the present invention was produced from hybridoma cells obtained by cell fusion technology from immunized mouse splenocytes using the mRID-uHAstalk antigen of influenza virus. , these were named 1G5 and 2C12. In another embodiment of the present invention, the monoclonal antibody of the present invention is produced from hybridoma cells obtained by cell fusion technology from mouse splenocytes immunized using the mRID-MERS RBD antigen of MERS coronavirus. Antibodies were prepared, and they were named mAb29 and mAb38, respectively.
본 발명은 또한, 상기 단일클론 항체 및/또는 다클론 항체를 포함하는 바이러스 감염 진단용 조성물을 제공한다.The present invention also provides a composition for diagnosing viral infection comprising the monoclonal antibody and/or polyclonal antibody.
상기 조성물은 유효성분으로 서열번호 4의 아미노산 서열로 이루어지는 uHAstalk에 특이적으로 결합하는 단일클론 항체, 서열번호 5의 아미노산 서열로 이루어지는 HAgd에 특이적으로 결합하는 다클론 항체, 또는 서열번호 6의 아미노산 서열로 이루어지는 MERS-RBD에 특이적으로 결합하는 단일클론 항체를 포함하며, 상기 항체를 이용하여 인플루엔자 바이러스 항원 또는 메르스 코로나 바이러스 항원에 대해 형성된 항원-항체 복합체를 검출함으로써 인플루엔자 바이러스를 면역학적으로 검출할 수 있다. 본 발명의 일 구현예에 의하면, 상기 서열번호 4의 아미노산 서열로 이루어진 펩타이드에 대한 항체는 A/Brisbane/59/2007 (IVR-148)(H1N1), A/California/07/09 (NYMC-X181)(H1N1), A/Michigan/45/2015 (NYMC-X275)(H1N1), A/Puerto Rico/8/34(H1N1), A/Singapore/GP1908/2015 (IVR-180)(H1N1), A/Singapore/01/57(H2N2), A/Anhui/01/05 (IBCDC-RG-6)(H5N1)등 인플루엔자 A 바이러스 그룹1 내 다양한 바이러스를 검출할 수 있다.The composition comprises a monoclonal antibody that specifically binds to uHAstalk consisting of the amino acid sequence of SEQ ID NO: 4 as an active ingredient, a polyclonal antibody that specifically binds to HAgd consisting of the amino acid sequence of SEQ ID NO: 5, or the amino acid of SEQ ID NO: 6 It includes a monoclonal antibody that specifically binds to MERS-RBD consisting of a sequence, and immunologically detects influenza virus by detecting an antigen-antibody complex formed against an influenza virus antigen or MERS coronavirus antigen using the antibody. can do. According to one embodiment of the present invention, the antibody against the peptide consisting of the amino acid sequence of SEQ ID NO: 4 is A/Brisbane/59/2007 (IVR-148)(H1N1), A/California/07/09 (NYMC-X181) )(H1N1), A/Michigan/45/2015 (NYMC-X275)(H1N1), A/Puerto Rico/8/34(H1N1), A/Singapore/GP1908/2015 (IVR-180)(H1N1), A Various viruses in influenza
또한, 본 발명은 상기 항체를 시료 샘플과 접촉시켜 형성된 항원-항체 복합체를 검출하는 단계를 포함하는 바이러스 검출방법을 제공한다.The present invention also provides a virus detection method comprising the step of detecting an antigen-antibody complex formed by contacting the antibody with a sample sample.
본 발명에서 용어 "항원-항체 복합체"란, 시료 중의 바이러스 항원과 이를 인지하는 본 발명에 따른 항체 또는 이의 절편의 결합물을 의미하며, 이러한 항원-항체 복합체는 비색법(colormetric method), 전기화학법(electrochemical method), 형광법(fluorimetric method), 발광법(luminometry), 입자계수법(particle counting method), 육안측정법(visual assessment) 및 섬광계수법(scintillation counting method)으로 이루어진 군에서 선택되는 임의의 방법으로 검출할 수 있다. 그러나 반드시 이들로만 제한되지 않고 다양한 응용과 적용이 가능하다.As used herein, the term "antigen-antibody complex" refers to a combination of a viral antigen in a sample and an antibody or fragment thereof according to the present invention that recognizes it, and the antigen-antibody complex is a colormetric method, an electrochemical method Detection by any method selected from the group consisting of electrochemical method, fluorimetric method, luminometry, particle counting method, visual assessment and scintillation counting method can do. However, it is not necessarily limited to these, and various applications and applications are possible.
본 발명에서는 항원-항체 복합체를 검출하기 위한 것으로 여러가지 표지체를 사용할 수 있다. 구체적인 예로는 효소, 형광물, 리간드, 발광물, 미소입자, 방사성 동위원소로 이루어진 그룹 중에서 선택될 수 있으며, 반드시 이들로만 한정되는 것은 아니다.In the present invention, various markers can be used to detect the antigen-antibody complex. Specific examples may be selected from the group consisting of enzymes, fluorescent substances, ligands, luminescent substances, microparticles, and radioactive isotopes, but are not necessarily limited thereto.
본 발명의 바이러스 검출방법에 있어서, 항원-항체 복합체를 검출하는 방법은 바람직하게는 효소면역흡착법(ELISA)을 이용하여 검출할 수 있으나, 이에 한정하는 것은 아니다. 효소면역흡착법은 고체 지지체에 부착된 항원을 인지하는 항체를 이용하는 직접적 ELISA, 고체 지지체에 부착된 항원을 인지하는 항체의 복합체에서 포획 항체를 인지하는 표지된 이차 항체를 이용하는 간접적 ELISA, 고체 지지체에 부착된 항체와 항원의 복합체에서 항원을 인지하는 표지된 또 다른 항체를 이용하는 직접적 샌드위치 ELISA, 고체 지지체에 부착된 항체와 항원의 복합체에서 항원을 인지하는 또 다른 항체와 반응시킨 후 이 항체를 인지하는 표지된 이차 항체를 이용하는 간접적 샌드위치 ELISA 등 다양한 ELISA 방법을 포함한다.In the virus detection method of the present invention, the method for detecting the antigen-antibody complex may preferably be detected using an enzyme immunosorbent assay (ELISA), but is not limited thereto. The enzyme immunosorbent method is a direct ELISA using an antibody recognizing an antigen attached to a solid support, indirect ELISA using a labeled secondary antibody recognizing a capture antibody in a complex of an antibody recognizing an antigen attached to a solid support, and attaching to a solid support. Direct sandwich ELISA using an antigen-recognizing antigen-recognizing antibody in a complex of an antibody and an antigen; direct sandwich ELISA using an antigen-recognizing antibody in a complex of an antibody attached to a solid support; It includes various ELISA methods such as indirect sandwich ELISA using a secondary antibody.
상기 단일클론항체 또는 이의 절편은 검출 표지체를 가질 수 있으며, 검출 표지체를 가지지 않을 경우는 이들 단일클론 항체 또는 이의 절편을 포획할 수 있고, 검출 표지체를 가지는 또 다른 항체를 처리하여 확인할 수 있다.The monoclonal antibody or fragment thereof may have a detection label, and when it does not have a detection label, these monoclonal antibodies or fragments thereof can be captured, and can be confirmed by treatment with another antibody having a detection label. have.
이하의 실시예는 발명의 이해를 돕기 위해 제시한 것으로 하기 내용이 발명의 보호범위를 한정하는 것은 아니다.The following examples are presented to help the understanding of the invention, and the following contents do not limit the protection scope of the invention.
생산 숙주 유래 RID 융합 항원에 대한 항체 생산Production of antibodies against RID fusion antigens from production hosts
본 발명에서 소개된 RID를 이용하여 항원을 생산하고 최종적인 항체를 생산하는 방법의 전체적 흐름은 도1A에 나타나 있다. 도 1은 마우스를 생산 숙주로 하여 마우스에서 유래된 RID를 융합 단백질로 이용한 재조합 항원의 생산과 이에 대한 단일클론항체 및 다클론항체를 제작하는 순서를 보여주고 있다. RID는 생산 숙주에 따라 적절한 종류로 선택하여 사용한다. RID는 다양한 생산 숙주에 널리 존재하고 있으며 그 중 도1B에서 쥐, 토끼, 인간의 RID 부분의 서열을 보여주고 있다. 항체의 생산은 기존에 알려진 hybridoma fusion 방법 이외 phage-display 등 다양한 방법이 적용될 수 있다.The overall flow of the method for producing antigen and final antibody using the RID introduced in the present invention is shown in Figure 1A. 1 shows the production of a recombinant antigen using a mouse-derived RID as a fusion protein using a mouse as a production host and a sequence of preparing monoclonal and polyclonal antibodies therefor. RID is used by selecting an appropriate type according to the production host. RIDs are widely present in various production hosts, and among them, the sequences of RID portions of mice, rabbits, and humans are shown in FIG. 1B. Various methods such as phage-display can be applied to the production of antibodies in addition to the known hybridoma fusion method.
서열번호 4의 유니버설 HA stalk 항원은 기존에 NCBI(National Center for Biotechnology Information)의 Influenza virus resource 데이터 베이스 내 존재하는 수천 개의 헤마글루티닌(HA) 단백질 서열정보를 분석하여 각 아미노산 서열 위치에 가장 여러 바이러스 주에서 빈번히 관찰되는 아미노산을 특정하여 이들을 모아 디자인한 새로운 형태의 인플루엔자 유니버설 항원이다. 디자인된 아미노산 서열은 이에 맞는 DNA 서열로 변환한 후 In vitro 에서의 핵산 합성을 통해 DNA를 제작하였다.The universal HA stalk antigen of SEQ ID NO: 4 analyzed the sequence information of thousands of hemagglutinin (HA) proteins existing in the Influenza virus resource database of the National Center for Biotechnology Information (NCBI), and the most numerous at each amino acid sequence position. It is a novel type of influenza universal antigen designed by collecting and designing amino acids frequently observed in virus strains. After the designed amino acid sequence was converted into a suitable DNA sequence, DNA was prepared through nucleic acid synthesis in vitro.
서열번호 5의 인플루엔자 HAgd 항원은 인플루엔자 A/Puerto Rico/08/1934 바이러스의 헤마글루티닌 항원 중 구조 상 globular domain으로 예측되는 부분만 선정하여 In vitro 에서의 핵산 합성을 통해 DNA를 제작하였다. Influenza HAgd antigen of SEQ ID NO: 5 was prepared by in vitro nucleic acid synthesis by selecting only a portion predicted to be a globular domain from among hemagglutinin antigens of influenza A/Puerto Rico/08/1934 virus.
서열번호 6의 MERS S1 항원 RBD는 MERS 바이러스의 S1 항원 중 구조상 Receptor binding domain 으로 예측되는 부분만 선정하여 In vitro 에서의 핵산 합성을 통해 DNA를 제작하였다.For the MERS S1 antigen RBD of SEQ ID NO: 6, only a portion predicted as a structural receptor binding domain among the S1 antigens of MERS virus was selected and DNA was prepared through nucleic acid synthesis in vitro.
유전자 클로닝-재조합 벡터의 제작Gene cloning - construction of recombinant vectors
본 발명에 사용된 발현벡터는 pGE-RID(4) 벡터이다. RID 융합 벡터는 T7 프로모터의 조절을 받으며 N-말단에 RID가 위치하고 이어 아스파르트산(Asp, D) 6개가 반복된 연결부분, TEV 프로테아제 인식서열, 다중 클로닝 부위(MCS; KpnI, BamHI, EcoRI, SalI, HindIII), 마지막으로 C-말단에 정제를 위한 6X 히스티딘(Histidine, His) tag 를 가지고 있다. 클로닝하고자 하는 목적단백질의 유전자는 다중 클로닝 부위(MCS; KpnI, BamHI, EcoRI, SalI, HindIII) 내 적절한 제한효소 2종의 절단 자리 사이에 삽입한다. 도2A, 도6A, 도8A, 도9A에서 각 목적 단백질의 유전자들은 중합효소 연쇄반응(PCR)을 통해 증폭된 이후 다중 클로닝 부위의 절단 자리에 삽입되었다. The expression vector used in the present invention is a pGE-RID(4) vector. The RID fusion vector is controlled by the T7 promoter, and RID is located at the N-terminus, followed by a linking portion with 6 repeated aspartic acids (Asp, D), TEV protease recognition sequence, multiple cloning site (MCS; KpnI, BamHI, EcoRI, SalI) , HindIII), and finally, a 6X histidine (His) tag for purification at the C-terminus. The gene of the target protein to be cloned is inserted between the cleavage sites of two appropriate restriction enzymes in the multiple cloning site (MCS; KpnI, BamHI, EcoRI, SalI, HindIII). 2A, 6A, 8A, and 9A, the genes of each target protein were amplified through polymerase chain reaction (PCR) and then inserted at the cleavage site of the multiple cloning site.
대장균 세포 내에서 RID융합 재조합 단백질의 수용성 발현 확인Confirmation of soluble expression of RID fusion recombinant protein in E. coli cells
재조합 단백질의 발현 양상을 보기 위한 실험은 15ml LB(Lysogeny broth)배지 에서의 배양된 대장균 내 단백질을 분석함으로써 이루어진다. 재조합 단백질은 대장균 숙주 BL21*(DE3)pLysS (Thermo Fisher)에서 과발현한다. 최초 배양은 암피실린 50μg/ml과 클로로암페니콜 30 μg/ml 이 포함된 3ml LB에서 37℃로 12시간 이상 배양 후 이중 500μl를 새 15ml LB배지에 옮겨 적정하여 OD600nm 가 0.5~0.7에 도달할 때까지 37℃에서 배양한다. 적정 OD600nm 값에 도달하면 다양한 온도 (예를 들어, 20℃, 16℃) 에서 1mM IPTG 를 첨가 후 적정 시간 배양한다. 배양이 끝난 배지는 3000g에서 원심분리를 통해 대장균 세포를 채취하며 이를 초음파 분해를 통해 세포 용해물을 얻어낸다. 얻어진 최초 세포 용해물내 전체 단백질을 T(Total)로 분류하며 세포용해물을 12,000g에서 원심분리를 통해 상등액과 침전된 부분으로 분리한다. 상등액 내 수용성 단백질은 S(Soluble)로 분류하며 침전된 부분내 불수용성 단백질은 P(Pellet)으로 분류한다(도2B, 도6B, 도8B, 도9B, 도9C). The experiment to see the expression pattern of the recombinant protein is made by analyzing the protein in E. coli cultured in 15ml LB (Lysogeny broth) medium. The recombinant protein is overexpressed in the E. coli host BL21*(DE3)pLysS (Thermo Fisher). The initial culture was carried out in 3 ml LB containing 50 μg/ml of ampicillin and 30 μg/ml of chloroamphenicol at 37° C. for more than 12 hours. Then, 500 μl of this was transferred to a new 15 ml LB medium and titrated to reach an OD 600 nm of 0.5-0.7. Incubate at 37°C until When an appropriate OD value of 600 nm is reached, 1 mM IPTG is added at various temperatures (eg, 20° C., 16° C.) and incubated for an appropriate time. E. coli cells are collected through centrifugation at 3000 g of the cultured medium, and a cell lysate is obtained through sonication. The total protein in the obtained first cell lysate is classified as T (Total), and the cell lysate is separated into a supernatant and a precipitated portion through centrifugation at 12,000 g. The water-soluble protein in the supernatant is classified as S (Soluble), and the water-insoluble protein in the precipitated portion is classified as P (Pellet) (Fig. 2B, Fig. 6B, Fig. 8B, Fig. 9B, Fig. 9C).
도 2B에서 발현된 목적단백질 mRID-uHAstalk은 37℃ 발현 시 20℃ 보다 발현량이 더 높았으나 전체 목적 단백질 중 수용성으로 발현된 정도는 20℃에서 더 높았다. 37℃에서는 전체 목적단백질 중 약 20%가 수용성이었으나 20℃에서는 전체 중 약 40%가 수용성이었다. 하지만 도 2C에서 RID를 융합하지 않은 uHAstalk 단백질의 발현 결과를 보았을 때 37℃와 20℃에서 모두 매우 미미한 발현량을 보였으며 대부분이 불수용성으로 발현되었다.The expression level of the target protein mRID-uHAstalk expressed in FIG. 2B was higher at 37°C than at 20°C, but the degree of soluble expression of the total target protein was higher at 20°C. At 37°C, about 20% of the total target protein was water-soluble, but at 20°C, about 40% of the total protein was water-soluble. However, when looking at the expression results of the uHAstalk protein not fused with RID in FIG. 2C, the expression levels were very insignificant at both 37°C and 20°C, and most of them were expressed as insoluble.
도 6B에서는 인플루엔자 A/Puerto Rico/08/34(H1N1) HAgd에 대한 발현 양상을 보여주었다. 20℃ 발현 시 RID를 융합하지 않은 HAgd는 대부분 불수용성으로 발현되었으나 세 종의 RID를 융합한 HAgd는 모두 증가한 수용성 정도를 보였다. 전체 발현된 목적단백질 중 mRID-HAgd의 경우 약 40%, rRID-HAgd의 경우 약 45%, hRID-HAgd의 경우 약 35%의 단백질이 수용성으로 발현되었다.Figure 6B showed the expression pattern for influenza A / Puerto Rico / 08 / 34 (H1N1) HAgd. When expressed at 20°C, most of the HAgd not fused with RID was expressed as water-insoluble, but all HAgd fused with three kinds of RID showed increased water solubility. Of the total expressed target proteins, about 40% of mRID-HAgd, about 45% of rRID-HAgd, and about 35% of hRID-HAgd were expressed in a water-soluble manner.
도 8B에서는 EGFP 단백질의 발현 양상을 보여주었다. RID를 융합하지 않은 EGFP는 전체 단백질 중 약 40%가 수용성으로 발현이 되었다. mRID-EGFP의 경우 전체 중 약 47% 가 수용성으로 발현되어 그 수용성 정도는 RID융합에도 크게 변하지는 않았으나 mRID 융합 시 발현량이 증가하는 모습을 보였다.Figure 8B showed the expression pattern of the EGFP protein. About 40% of the total protein of EGFP not fused with RID was expressed as water soluble. In the case of mRID-EGFP, about 47% of the total was expressed as water soluble, and the degree of water solubility did not change significantly even with RID fusion, but the expression level increased during mRID fusion.
도9B, C에서는 MERS 바이러스의 RBD 발현양상을 보여주었다. RID 융합하지 않은 RBD의 경우 대부분 불수용성으로 발현됨을 볼 수 있었으나 mRID-MERS RBD의 경우 다양한 온도에서 발현 결과 증가된 수용성을 보였다. 37℃ 발현 시 약 58%, 30℃ 발현 시 약 67%, 20℃ 발현 시 약 68%, 16℃ 발현 시 거의 100% 가 수용성으로 발현되었다.9B, C shows the RBD expression pattern of the MERS virus. In the case of RBD not fused to RID, it was found that most of the expression was insoluble, but in the case of mRID-MERS RBD, the expression at various temperatures showed increased water solubility. At 37°C, about 58%, at 30°C, about 67%, at 20°C, about 68%, and at 16°C, almost 100% was expressed as water-soluble.
RID융합 재조합단백질의 정제Purification of RID fusion recombinant protein
RID 융합 재조합단백질들은 니켈 친화성 크로마토그래피를 이용해 정제하였다. 각각의 이미다졸(Imidazole)의 농도 조절을 위해 A 버퍼 [50 mM Tris-Cl(pH 7.5), 300 mM 염화 나트륨, 10% 글리세롤, 2mM 2-머캡토에탄올, Tween-20 0.05%, and 10mM 이미다졸] 과 B 버퍼 [50 mM Tris-Cl(pH 7.5), 300 mM 염화 나트륨, 10% 글리세롤, 2mM 2-머캡토에탄올, Tween-20 0.05%, and 300mM 이미다졸]을 적절한 비율로 섞으며 직선형으로 이미다졸 농도를 점차적으로 증가시켜 Ni-NTA 컬럼 레진(GE Healthcare Life Sciences, Little Chalfont, Buckinghamshire, UK)에 넣음으로써 RID융합 단백질을 정제한다.RID fusion recombinant proteins were purified using nickel affinity chromatography. To control the concentration of each imidazole, buffer A [50 mM Tris-Cl (pH 7.5), 300 mM sodium chloride, 10% glycerol, 2 mM 2-mercaptoethanol, Tween-20 0.05%, and 10 mM imidazole) Dazole] and B buffer [50 mM Tris-Cl (pH 7.5), 300 mM sodium chloride, 10% glycerol, 2 mM 2-mercaptoethanol, Tween-20 0.05%, and 300 mM imidazole] in appropriate proportions were mixed in a straight line. The RID fusion protein is purified by gradually increasing the imidazole concentration with Ni-NTA column resin (GE Healthcare Life Sciences, Little Chalfont, Buckinghamshire, UK).
mRID 융합 유니버설 헤마글루티닌 stalk 도메인(mRID-uHAstalk)에 대한 단일클론항체 생산 및 평가Production and evaluation of monoclonal antibody against mRID fusion universal hemagglutinin stalk domain (mRID-uHAstalk)
정제된 mRID-uHAstalk 단백질은 쥐에 접종된 후 기존에 알려진 하이브리도마 융합 방법(28, 29)을 통하여 접종된 단백질에 특이적인 클론을 선정하여 항체 정제를 진행하였다. 6주령 BALB/c 쥐에 mRID-uHAstalk 항원을 2주 간격으로 2회 접종하며 면역보강제(Adjuvant)로 프로이드 어주번트(Freund's adjuvant)를 사용한다. 2차 접종 3일 후 비장 세포를 채취하며 널리 하이브리도마 융합 방법(28, 29)을 이용하여 하이브리도마 세포를 제작한다. 면역된 항원에 대한 포지티브 클론(positive clone)은 해당 항원에 대한 ELISA를 통해 스크리닝 한다.After the purified mRID-uHAstalk protein was inoculated into mice, a clone specific for the inoculated protein was selected through the known hybridoma fusion method (28, 29) and antibody purification was performed. 6-week-old BALB/c mice are inoculated with mRID-uHAstalk antigen twice at an interval of 2 weeks, and Freund's adjuvant is used as an adjuvant. Splenocytes are harvested 3 days after the second inoculation, and hybridoma cells are prepared using the widely used hybridoma fusion method (28, 29). Positive clones for the immunized antigen are screened through ELISA for the antigen.
도 3A는 쥐에 면역 후 제작한 포지티브 클론 15종에 대하여 접종 항원인 mRID-uHA stalk과 융합단백질인 mRID에 대한 ELISA 결과 놀랍게도 15개의 모든 클론에서 접종항원에 대해서만 반응성을 보이며 반대로 융합파트너인 mRID에 대하서는 반응을 보이지 않았다. 이는 mRID가 쥐에는 자가항원으로서 항원성을 보이지 않음을 보여준다. 따라서 RID를 이용한 융합항원 생산방법은 융합파트너를 제거하지 않고 융합단백질의 형태로 접종하여도 높은 효율로 목적단백질 특이적 클론 생산이 가능함을 보여준다.3A shows the results of ELISA for the inoculation antigen mRID-uHA stalk and the fusion protein mRID for 15 positive clones prepared after immunization with mice. Surprisingly, all 15 clones showed reactivity only to the inoculation antigen. Daeha did not respond. This shows that mRID does not show antigenicity as an autoantigen in mice. Therefore, the fusion antigen production method using RID shows that it is possible to produce a target protein-specific clone with high efficiency even when inoculated in the form of a fusion protein without removing the fusion partner.
최종 선정된 클론은 Protein G resin (GE Healthcare, Little Chalfont, UK)을 통해 정제한다. 상기 실시예에서는 mRID-uHAstalk에 대한 단일클론항체 1G5와 2C12를 최종 생산하였다. 위 항체 생산은 에이티젠(경기도 성남, 대한민국)에서 이루어졌다. 1G5와 2C12는 접종된 단백질(mRID-uHAstalk)과 융합단백질(mRID)에 대한 ELISA를 통해 유니버설 HA stalk domain 특이적 항체인지 검증하였다. 도 3B는 1G5를 이용한 ELISA 결과이며 도 3C는 2C12를 이용한 ELISA 결과이다. 위 결과 두 단일클론항체는 융합파트너인 mRID에는 결합하지 않았으나 mRID-uHAstalk에는 결합하였다. 따라서 두 단일클론항체는 uHAstalk 항원에 특이적인 항체임을 알 수 있었으며 mRID는 쥐에 접종되어 항체를 유도하지 않음을 다시 한번 보여주었다.The final selected clones are purified through Protein G resin (GE Healthcare, Little Chalfont, UK). In the above example, monoclonal antibodies 1G5 and 2C12 against mRID-uHAstalk were finally produced. The above antibody production was made in ATIGEN (Seongnam, Gyeonggi-do, Korea). 1G5 and 2C12 were verified as universal HA stalk domain-specific antibodies through ELISA for inoculated protein (mRID-uHAstalk) and fusion protein (mRID). 3B is an ELISA result using 1G5, and FIG. 3C is an ELISA result using 2C12. As a result, the two monoclonal antibodies did not bind to mRID, a fusion partner, but bound to mRID-uHAstalk. Therefore, it was confirmed that the two monoclonal antibodies were specific for the uHAstalk antigen, and once again showed that mRID did not induce antibodies when inoculated into mice.
생산된 단일클론항체는 여러 종류의 인플루엔자 바이러스 HA 단백질에 대한 ELISA를 수행함으로써 항체가 실제 타겟과 잘 결합하는지를 평가하였다. 본 ELISA에 사용된 HA 단백질은 영국 NIBSC(National Institute for Biological Standards and Control)에서 공급된 인플루엔자 표준 HA 항원이며 사용된 바이러스 종은 A/Brisbane/59/2007 (IVR-148)(H1N1), A/California/07/09 (NYMC-X181)(H1N1), A/Michigan/45/2015 (NYMC-X275)(H1N1), A/Puerto Rico/8/34(H1N1), A/Singapore/GP1908/2015 (IVR-180)(H1N1), A/Singapore/01/57(H2N2), A/Anhui/01/05 (IBCDC-RG-6)(H5N1) 이다. ELISA 과정 중 HA stalk domain에 결합하는 항체인 1G5, 2C12의 항원 접근성 향상을 위하여 DTT(Dithiothreitol)처리를 통하여 HA의 head domain 과 stalk domain 사이의 disulfide bond를 제거하여 head domain을 제거하며 low pH 버퍼를 통해 단백질 구조를 약간 lose하게 해주는 항원 전처리를 진행하였다. 도 4A-G, 도 5A-G의 결과에 따르면 두 단일클론항체 1G5 및 2C12는 실제 야생형 인플루엔자 바이러스의 HA에 잘 결합하는 것으로 보인다. 따라서 두 단일클론항체는 실제 HA 단백질 항원에 결합할 수 있음을 확인하였다. 또한 위 결과를 통해 단일클론항체 생산에 사용된 mRID-uHAstalk 단백질이 올바른 구조를 이루었다고 생각 할 수 있다.The produced monoclonal antibody was evaluated whether the antibody binds well to the actual target by performing ELISA for several types of influenza virus HA protein. The HA protein used in this ELISA was an influenza standard HA antigen supplied by the National Institute for Biological Standards and Control (NIBSC), UK, and the virus species used were A/Brisbane/59/2007 (IVR-148)(H1N1), A/ California/07/09 (NYMC-X181)(H1N1), A/Michigan/45/2015 (NYMC-X275)(H1N1), A/Puerto Rico/8/34(H1N1), A/Singapore/GP1908/2015 ( IVR-180) (H1N1), A/Singapore/01/57 (H2N2), A/Anhui/01/05 (IBCDC-RG-6) (H5N1). During the ELISA process, to improve antigen accessibility of 1G5 and 2C12, which are antibodies that bind to the HA stalk domain, DTT (Dithiothreitol) treatment is used to remove the disulfide bond between the head domain and the stalk domain of HA to remove the head domain and use a low pH buffer. Antigen pretreatment was performed to slightly lose the protein structure. According to the results of FIGS. 4A-G and 5A-G, the two monoclonal antibodies 1G5 and 2C12 seem to bind well to the HA of the actual wild-type influenza virus. Therefore, it was confirmed that the two monoclonal antibodies could actually bind to the HA protein antigen. Also, from the above results, it can be considered that the mRID-uHAstalk protein used for monoclonal antibody production has the correct structure.
mRID 융합 HA globular domain (mRID-HAgd)의 단백질 접힘 적합성 확인Confirmation of protein folding suitability of mRID fusion HA globular domain (mRID-HAgd)
본 실시예는 인플루엔자 A/Puerto Rico/8/34 바이러스의 HA 단백질이 mRID 융합을 통해 적절한 구조를 잘 이루어 생산되었는지 검증하기 위한 실험에 대한 내용이다. 실제 인플루엔자 A/Puerto Rico/8/34 바이러스를 쥐에 접종하여 항혈청을 얻었다. 얻어낸 항혈청에는 해당 바이러스의 HA단백질 항원에 대한 항체들이 포함되어 있다. 이러한 쥐의 항혈청을 1차 항체로 사용하여 mRID-HAgd, rRID-HAgd 단백질에 대한 ELISA와 Western blot을 수행하였다. 이러한 결과는 도 7A 및 도 7B에 나타나 있다. 도 7A 에서의 ELISA 결과는 바이러스를 접종한 항혈청 내 항체가 mRID-HAgd에는 잘 결합하나 mRID, mRID가 융합되지 않은 HAgd에 대해서는 잘 결합하지 않는 모습을 보인다. 또한 HAgd는 RID 융합 없이는 대장균 내에서 올바른 구조로 생산되지 않음을 보여주었다. 이에 더해 Western blot 결과에서는 상기 ELISA 결과와 일맥상통하는 결과를 보이며 이에 더해 TEV 프로테아제를 통해 mRID-HAgd에서 mRID를 잘라낸 HAgd에도 항체에 잘 결합하는 것으로 보인다. 이를 통해 mRID 융합을 통해 일단 HAgd가 생산되면 이후에 mRID를 잘라내도 HAgd의 올바른 구조가 유지되는 것을 확인할 수 있다. 도 7B 에서는 쥐의 항혈청 내 항체가 rRID-HAgd에는 잘 결합하나 rRID, rRID가 융합되지 않은 HAgd에 대해서는 잘 결합하지 않는 것으로 보였다. 이를 통해 rRID의 융합으로 HAgd가 올바른 구조를 이루어 생산되었음을 알 수 있다. 또한 Western blot 결과에서도 위 ELISA와 유사한 결과를 보임과 더하여 TEV 프로테아제를 통해 rRID-HAgd에서 rRID를 잘라낸 HAgd에도 항체 잘 결합하는 것으로 보인다. 따라서 RID-HAgd의 구조가 실제 바이러스의 HAgd의 구조와 유사함을 확인할 수 있었다. 또한 도 7C 에서는 다양한 바이러스를 쥐에 접종하여 얻은 혈청을 이용하여 mRID-HAgd에 대한 ELISA를 진행한 결과 A/Puerto Rico/8/34 접종 한 쥐의 항혈청에서만 특이적 반응이 있는 것으로 보인다. 이를 통해 mRID-HAgd의 구조가 종(strain) 특이적 항원을 구분 가능할 수 있게 그 본래의 구조를 완벽히 이루고 있음을 보여준다.This example is about an experiment to verify that the HA protein of influenza A/Puerto Rico/8/34 virus was produced with an appropriate structure through mRID fusion. Antisera were obtained by inoculating mice with actual influenza A/Puerto Rico/8/34 virus. The obtained antiserum contains antibodies to the HA protein antigen of the virus. ELISA and Western blot for mRID-HAgd and rRID-HAgd proteins were performed using the rat antiserum as a primary antibody. These results are shown in Figures 7A and 7B. The ELISA result in FIG. 7A shows that the antibody in the antiserum inoculated with the virus binds well to mRID-HAgd, but does not bind well to HAgd to which mRID and mRID are not fused. We also showed that HAgd is not produced in the correct conformation in E. coli without RID fusion. In addition, Western blot results show results consistent with the above ELISA results, and in addition, it appears that HAgd, which is mRID-cleaved from mRID-HAgd through TEV protease, also binds well to the antibody. Through this, it can be confirmed that once HAgd is produced through mRID fusion, the correct structure of HAgd is maintained even after mRID is cut. In FIG. 7B , it was shown that the mouse antiserum antibody binds well to rRID-HAgd, but does not bind well to HAgd to which rRID and rRID are not fused. Through this, it can be seen that HAgd was produced with the correct structure through the fusion of rRID. In addition, Western blot results showed similar results to the above ELISA, and in addition, the antibody seemed to bind well to HAgd, which was cut from rRID-HAgd by TEV protease. Therefore, it was confirmed that the structure of RID-HAgd was similar to that of HAgd of the actual virus. In addition, in FIG. 7C , as a result of ELISA for mRID-HAgd using serum obtained by inoculating mice with various viruses, it seems that only the antisera of mice inoculated with A/Puerto Rico/8/34 had a specific reaction. This shows that the structure of mRID-HAgd perfectly forms its original structure to be able to distinguish a strain-specific antigen.
mRID 및 rRID 융합 HA globular domain (mRID-HAgd)에 대한 다클론항체를 포함한 항혈청의 생산 및 평가Production and evaluation of antisera including polyclonal antibodies against mRID and rRID fusion HA globular domain (mRID-HAgd)
mRID-HAgd에 대한 쥐 항혈청은 상기 항원을 6주령 BALB/c 쥐에 접종하였다. 20μg의 mRID-HAgd 단백질은 50μg의 백반 보충제(Thermo Fisher)을 섞어 접종하며 2주 간격으로 총 3회 복강내로 접종된다. 실험에 사용된 항혈청은 3차 접종 후 2주 뒤에 안와채혈을 통해 얻어졌다.For murine antisera against mRID-HAgd, the antigen was inoculated into 6-week-old BALB/c mice. 20μg of mRID-HAgd protein is inoculated with 50μg of alum supplement (Thermo Fisher), and inoculated intraperitoneally 3 times at intervals of 2 weeks. The antiserum used in the experiment was obtained through
rRID-HAgd에 대한 토끼 항혈청은 상기 항원 1mg을 프로이드 어주번트(Freund's adjuvant)와 함께 섞어 피내 주사로 접종하였다. 최초 접종 4주 후 2주 간격으로 2번 추가 접종하였으며 최종 접종 일주일 후 혈청을 채취하였다. 본 실험은 영인프론티어(서울시 가산동, 대한민국)에서 수행되었다.The rabbit antiserum against rRID-HAgd was inoculated by intradermal injection by mixing 1 mg of the antigen with Freund's adjuvant. Four weeks after the initial inoculation, two additional inoculations were performed at an interval of two weeks, and serum was collected one week after the last inoculation. This experiment was conducted at Youngin Frontier (Gasan-dong, Seoul, Korea).
도 7D는 mRID-HAgd에 대한 쥐 항혈청을 이용하여 융합파트너 mRID, mRID-HAgd에 대한 ELISA와 Western blot을 수행한 결과이다. ELISA 결과는 항혈청 내 항체가 mRID-HAgd에는 잘 결합하나 mRID에는 상대적으로 잘 결합하지 않는 모습을 보인다. 이에 더해 Western blot 결과에서는 mRID-HAgd와 TEV 프로테아제를 통해 mRID-HAgd에서 mRID를 잘라낸 HAgd에는 항체가 잘 결합하는 것으로 보였다. 하지만 mRID에는 결합하지 않았다. 이를 통해 혈청 내 mRID-HAgd 항체들은 HAgd에만 특이적이며 융합파트너 mRID에는 잘 결합하지 않는 것으로 보인다.7D shows the results of ELISA and Western blot for fusion partners mRID and mRID-HAgd using rat antiserum against mRID-HAgd. ELISA results show that the antibody in the antiserum binds well to mRID-HAgd but does not bind well to mRID. In addition, Western blot results showed that the antibody was well bound to mRID-HAgd and HAgd, which was cut from mRID-HAgd by TEV protease. However, it did not bind to mRID. From this, it seems that mRID-HAgd antibodies in the serum are specific only to HAgd and do not bind well to the fusion partner mRID.
도 7E는 rRID-HAgd에 대한 토끼 혈청을 이용하여 융합파트너 rRID, rRID-HAgd에 대한 ELISA와 Western blot을 수행한 결과이다. ELISA 결과는 항혈청 내 항체가 rRID-HAgd에는 잘 결합하나 rRID에는 상대적으로 잘 결합하지 않는 모습을 보인다. 이에 더해 Western blot 결과에서는 rRID-HAgd와 TEV 프로테아제를 통해 rRID-HAgd에서 rRID를 잘라낸 HAgd에는 항체가 잘 결합하는 것으로 보였다. 하지만 rRID에는 결합하지 않았다. 이를 통해 항혈청 내 rRID-HAgd 항체들은 HAgd에만 특이적이며 융합파트너 rRID에는 잘 결합하지 않는 것으로 보인다.7E shows the results of ELISA and Western blot for fusion partners rRID and rRID-HAgd using rabbit serum for rRID-HAgd. ELISA results show that the antibody in the antiserum binds well to rRID-HAgd but does not bind well to rRID. In addition, Western blot results showed that the antibody was well bound to rRID-HAgd and HAgd, which was cut from rRID-HAgd by TEV protease. However, it did not bind to rRID. From this, it seems that the rRID-HAgd antibodies in the antiserum are specific only to HAgd and do not bind well to the fusion partner rRID.
도 7F는 E.coli의 LysRS-HAgd에 대한 쥐 항혈청을 이용하여 융합단백질 LysRS, LysRS-HAgd에 대한 ELISA를 수행한 결과이다. 그 결과 해당 항혈청은 LysRS와 LysRS-HAgd에 모두 반응하는 것으로 보였다. 이는 융합 단백질인 LysRS에 대한 면역반응이 목적단백질에 대한 면역반응을 압도하여 융합단백질에 대한 항체만 생산되었음을 보여준다. 따라서 LysRS가 융합파트너로서 목적항원의 수용성 생산에는 좋은 기능을 할 수는 있으나 이에 대한 항체 생산의 경우에는 적합하지 않음을 보여주고 있다.7F shows the results of performing ELISA for LysRS and LysRS-HAgd fusion proteins using mouse antiserum against LysRS-HAgd of E. coli. As a result, the antisera appeared to respond to both LysRS and LysRS-HAgd. This shows that the immune response to LysRS, a fusion protein, overwhelms the immune response to the target protein, and only antibodies to the fusion protein were produced. Therefore, it is shown that LysRS as a fusion partner can function well for the water-soluble production of the target antigen, but is not suitable for the production of antibodies against it.
mRID 및 rRID 융합 EGFP (mRID-EGFP)에 대한 다클론항체를 포함한 항혈청의 생산과 평가Production and evaluation of antisera including polyclonal antibodies against mRID and rRID fusion EGFP (mRID-EGFP)
mRID-EGFP에 대한 쥐 항혈청은 상기 항원을 6주령 BALB/c 쥐에 접종하였다. mRID-EGFP 단백질은 백반 보충제(Thermo Fisher)을 섞어 접종하며 2주 간격으로 총 3회 복강 내로 접종된다. 실험에 사용된 항혈청은 3차 접종 후 2주 뒤에 안와채혈을 통해 얻어졌다.For murine antisera against mRID-EGFP, the antigen was inoculated into 6-week-old BALB/c mice. The mRID-EGFP protein is inoculated with alum supplement (Thermo Fisher) and inoculated intraperitoneally 3 times at 2 week intervals. The antiserum used in the experiment was obtained through
rRID-EGFP에 대한 토끼 항혈청은 상기 항원 1mg을 프로이드 어주번트(Freund's adjuvant)와 함께 섞어 피내 주사로 접종하였다. 최초 접종 4주 후 2주 간격으로 2번 추가 접종하였으며 최종 접종 일주일 후 혈청을 채취하였다. 본 실험은 영인프론티어(서울시 가산동, 대한민국)에서 수행되었다.The rabbit antiserum against rRID-EGFP was inoculated by intradermal injection by mixing 1 mg of the antigen with Freund's adjuvant. Four weeks after the initial inoculation, two additional inoculations were performed at an interval of two weeks, and serum was collected one week after the last inoculation. This experiment was conducted at Youngin Frontier (Gasan-dong, Seoul, Korea).
도 8C는 mRID-EGFP에 대한 쥐 항혈청을 이용하여 융합파트너 mRID, mRID-EGFP에 대한 ELISA와 Western blot을 수행한 결과이다. ELISA 결과는 항혈청 내 항체가 mRID-EGFP에는 잘 결합하나 mRID에는 상대적으로 잘 결합하지 않는 모습을 보인다. 이에 더해 Western blot 결과에서는 mRID-EGFP와 TEV 프로테아제를 통해 mRID-EGFP에서 mRID를 잘라낸 EGFP에는 항체가 잘 결합하는 것으로 보였다. 하지만 mRID에는 결합하지 않았다. 이를 통해 항혈청 내 mRID-EGFP 항체들은 EGFP에만 특이적이며 융합파트너 mRID에는 잘 결합하지 않는 것으로 보인다.8C shows the results of ELISA and Western blot for fusion partners mRID and mRID-EGFP using rat antiserum against mRID-EGFP. ELISA results show that the antibody in the antiserum binds well to mRID-EGFP but does not bind well to mRID. In addition, Western blot results showed that the antibody was well bound to EGFP, which was cut from mRID-EGFP by mRID-EGFP and TEV protease. However, it did not bind to mRID. From this, it seems that the mRID-EGFP antibodies in the antiserum are specific only to EGFP and do not bind well to the fusion partner mRID.
도 8D는 rRID-EGFP에 대한 토끼 혈청을 이용하여 융합파트너 rRID, rRID-EGFP에 대한 ELISA와 Western blot을 수행한 결과이다. ELISA 결과는 항혈청내 항체가 rRID-EGFP에는 잘 결합하나 rRID에는 상대적으로 잘 결합하지 않는 모습을 보인다. 이에 더해 Western blot 결과에서는 rRID-EGFP와 TEV 프로테아제를 통해 rRID-EGFP에서 rRID를 잘라낸 EGFP에는 항체가 잘 결합하는 것으로 보였다. 하지만 rRID에는 결합하지 않았다. 이를 통해 혈청 내 rRID-EGFP 항체들은 EGFP에만 특이적이며 융합파트너 rRID에는 잘 결합하지 않는 것으로 보인다.8D shows the results of ELISA and Western blot for fusion partners rRID and rRID-EGFP using rabbit serum for rRID-EGFP. ELISA results show that the antibody in the antiserum binds well to rRID-EGFP but does not bind well to rRID. In addition, Western blot results showed that the antibody was well bound to EGFP, which was cut from rRID-EGFP by rRID-EGFP and TEV protease. However, it did not bind to rRID. From this, it seems that the rRID-EGFP antibodies in the serum are specific only to EGFP and do not bind well to the fusion partner rRID.
mRID 융합 MERS S1 항원 RBD 접힘 적합성 확인Confirmation of mRID fusion MERS S1 antigen RBD folding suitability
본 실시예는 MERS S1 RBD단백질은 mRID 융합을 통해 생산하고 상기 단백질이 올바른 구조를 이루고 있는지 검증하기 위한 실험이다. 실제 MERS 바이러스의 RBD 단백질은 hDPP4(Human dipeptidyl peptidase4) 단백질과 결합한다고 알려져 있다(30, 31). 따라서 mRID-MERS RBD 단백질이 올바른 구조를 이루었는지 확인하기 위해 hDPP4와의 결합 여부를 확인해 보았다. 이를 위해 mRID-MERS RBD에 결합할 수 있는 Anti-6xHis tag antibody(Thermo Fisher)를 이용하여 Western blot과 ELISA를 통한 상호 면역 침전 법 분석(Co-immunoprecipitation analysis)을 진행하였다. 도 9D는 ELISA 결과로 mRID-MERS RBD는 293T 세포에서 생산된 RBD단백질과 유사한 정도로 hDPP4와 잘 결합하는 것으로 확인되었다. 따라서 위 결과에서도 mRID-MERS RBD가 올바른 구조를 이루고 있음을 보여주었다.This example is an experiment to verify that the MERS S1 RBD protein is produced through mRID fusion and the protein has a correct structure. It is known that the RBD protein of the MERS virus actually binds to the human dipeptidyl peptidase4 (hDPP4) protein (30, 31). Therefore, to check whether the mRID-MERS RBD protein had the correct structure, it was checked whether it binds to hDPP4. To this end, co-immunoprecipitation analysis was performed through Western blot and ELISA using an Anti-6xHis tag antibody (Thermo Fisher) capable of binding to mRID-MERS RBD. 9D shows the results of ELISA, it was confirmed that mRID-MERS RBD binds well to hDPP4 to a degree similar to the RBD protein produced in 293T cells. Therefore, the above results also showed that the mRID-MERS RBD had the correct structure.
mRID 융합 MERS RBD (mRID-MERS RBD)에 대한 단일클론항체 생산 및 평가Production and evaluation of monoclonal antibody against mRID fusion MERS RBD (mRID-MERS RBD)
정제된 mRID-MERS RBD 단백질은 쥐에 접종된 후 기존에 알려진 하이브리도마 융합 방법(28, 29)을 통하여 접종된 단백질에 특이적인 클론을 선정하여 항체 정제를 진행하였다. 6주령 BALB/c 쥐에 mRID-MERS RBD 항원을 2주 간격으로 2회 접종하며 면역보강제(Adjuvant)로 프로이드 어주번트(Freund's adjuvant)를 사용한다. 2차 접종 3일 후 비장 세포를 채취하며 널리 알려진 하이브리도마 융합 방법(28, 29)을 이용하여 하이브리도마 세포를 제작한다. 면역된 항원에 대한 포지티브 클론(positive clone)은 해당 항원에 대한 ELISA를 통해 스크리닝 한다. 최종 선정된 클론은 Protein G resin (GE Healthcare, Little Chalfont, UK)을 통해 정제한다. 상기 실시예에서는 mRID-MERS RBD에 대한 단일클론항체 mAb29와 mAb38을 생산하였다. 위 항원 생산은 에이티젠(경기도 성남, 대한민국)에서 이루어졌다.After the purified mRID-MERS RBD protein was inoculated into mice, a clone specific for the inoculated protein was selected through the known hybridoma fusion method (28, 29) and antibody purification was performed. 6-week-old BALB/c mice are inoculated with mRID-MERS RBD antigen twice at an interval of 2 weeks, and Freund's adjuvant is used as an adjuvant. Splenocytes are collected 3 days after the second inoculation, and hybridoma cells are prepared using the well-known hybridoma fusion method (28, 29). Positive clones for the immunized antigen are screened through ELISA for the antigen. The final selected clones are purified through Protein G resin (GE Healthcare, Little Chalfont, UK). In the above example, monoclonal antibodies mAb29 and mAb38 against mRID-MERS RBD were produced. The above antigen production was performed in ATIGEN (Seongnam, Gyeonggi-do, Korea).
도 9E에서는 mRID-MERS RBD에 대한 두 단일클론항체를 이용해 Sino Biological (베이징, 중국)에서 공급된 Baculo-insect expression에서 생산된 RBD 단백질에 대하여 ELISA를 수행한 결과이다. 상기 단백질에는 어떠한 융합단백질도 포함 되어있지 않다. ELISA 결과 두 단일클론항체는 RBD단백질과 결합할 수 있음을 알 수 있었다. 따라서 두 단일클론항체는 RBD 특이적 항체라고 할 수 있다.9E shows the results of performing ELISA on the RBD protein produced by Baculo-insect expression supplied from Sino Biological (Beijing, China) using two monoclonal antibodies against mRID-MERS RBD. The protein does not contain any fusion protein. As a result of ELISA, it was found that the two monoclonal antibodies could bind to the RBD protein. Therefore, both monoclonal antibodies can be said to be RBD-specific antibodies.
도 9F는 mRID-MERS RBD, mRID, 대장균 BL21*(DE3)pLysS (Thermo Fisher)competent cell 내재 단백질에 대하여 mAb29를 이용해 Western blot을 수행한 결과이다. 위 결과 mRID-MERS RBD에만 단일클론항체가 결합하였으며 mRID와 대장균 내재 단백질에 대해서는 결합하지 않는 것으로 보인다. 따라서 mRID는 쥐 내에서 면역을 유도하지 않는 융합파트너로 사용될 수 있음을 보였다.9F shows the results of Western blot using mAb29 for mRID-MERS RBD, mRID, E. coli BL21*(DE3)pLysS (Thermo Fisher) competent cell-intrinsic protein. As a result, the monoclonal antibody only bound to mRID-MERS RBD, but it seems that it did not bind to mRID and E. coli endogenous protein. Therefore, it was shown that mRID can be used as a fusion partner that does not induce immunity in mice.
<참고문헌><References>
1. Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975;256(5517):495-7.1. Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975;256(5517):495-7.
2. Singh S, Kumar N, Dwiwedi P, Charan J, Kaur R, Sidhu P, et al. Monoclonal Antibodies: A Review. Current clinical pharmacology. 2017.2. Singh S, Kumar N, Dwiwedi P, Charan J, Kaur R, Sidhu P, et al. Monoclonal Antibodies: A Review. Current clinical pharmacology. 2017.
3. Nahta R, Esteva FJ. Trastuzumab: triumphs and tribulations. Oncogene. 2007;26(25):3637-43.3. Nahta R, Esteva FJ. Trastuzumab: triumphs and tribulations. Oncogene. 2007;26(25):3637-43.
4. Lugering A, Schmidt M, Lugering N, Pauels HG, Domschke W, Kucharzik T. Infliximab induces apoptosis in monocytes from patients with chronic active Crohn's disease by using a caspase-dependent pathway. Gastroenterology. 2001;121(5):1145-57.4. Lugering A, Schmidt M, Lugering N, Pauels HG, Domschke W, Kucharzik T. Infliximab induces apoptosis in monocytes from patients with chronic active Crohn's disease by using a caspase-dependent pathway. Gastroenterology. 2001;121(5):1145-57.
5. Gurcan HM, Keskin DB, Stern JN, Nitzberg MA, Shekhani H, Ahmed AR. A review of the current use of rituximab in autoimmune diseases. International immunopharmacology. 2009;9(1):10-25.5. Gurcan HM, Keskin DB, Stern JN, Nitzberg MA, Shekhani H, Ahmed AR. A review of the current use of rituximab in autoimmune diseases. International immunopharmacology. 2009;9(1):10-25.
6. Keating GM. Bevacizumab: a review of its use in advanced cancer. Drugs. 2014;74(16):1891-925.6. Keating GM. Bevacizumab: a review of its use in advanced cancer. Drugs. 2014;74(16):1891-925.
7. Yamaguchi H, Miyazaki M. Refolding techniques for recovering biologically active recombinant proteins from inclusion bodies. Biomolecules. 2014;4(1):235-51.7. Yamaguchi H, Miyazaki M. Refolding techniques for recovering biologically active recombinant proteins from inclusion bodies. Biomolecules. 2014;4(1):235-51.
8. Yang Z, Zhang L, Zhang Y, Zhang T, Feng Y, Lu X, et al. Highly efficient production of soluble proteins from insoluble inclusion bodies by a two-step-denaturing and refolding method. PloS one. 2011;6(7):e22981.8. Yang Z, Zhang L, Zhang Y, Zhang T, Feng Y, Lu X, et al. Highly efficient production of soluble proteins from insoluble inclusion bodies by a two-step-denaturing and refolding method. PloS one. 2011;6(7):e22981.
9. Singh SM, Panda AK. Solubilization and refolding of bacterial inclusion body proteins. Journal of bioscience and bioengineering. 2005;99(4):303-10.9. Singh SM, Panda AK. Solubilization and refolding of bacterial inclusion body proteins. Journal of bioscience and bioengineering. 2005;99(4):303-10.
10. Basu A, Li X, Leong SS. Refolding of proteins from inclusion bodies: rational design and recipes. Applied microbiology and biotechnology. 2011;92(2):241-51.10. Basu A, Li X, Leong SS. Refolding of proteins from inclusion bodies: rational design and recipes. Applied microbiology and biotechnology. 2011;92(2):241-51.
11. Baneyx F. Recombinant protein expression in Escherichia coli. Current opinion in biotechnology. 1999;10(5):411-21.11. Baneyx F. Recombinant protein expression in Escherichia coli. Current opinion in biotechnology. 1999;10(5):411-21.
12. Esposito D, Chatterjee DK. Enhancement of soluble protein expression through the use of fusion tags. Current opinion in biotechnology. 2006;17(4):353-8.12. Esposito D, Chatterjee DK. Enhancement of soluble protein expression through the use of fusion tags. Current opinion in biotechnology. 2006;17(4):353-8.
13. Pryor KD, Leiting B. High-level expression of soluble protein in Escherichia coli using a His6-tag and maltose-binding-protein double-affinity fusion system. Protein expression and purification. 1997;10(3):309-19.13. Pryor KD, Leiting B. High-level expression of soluble protein in Escherichia coli using a His6-tag and maltose-binding-protein double-affinity fusion system. Protein expression and purification. 1997;10(3):309-19.
14. Fox JD, Routzahn KM, Bucher MH, Waugh DS. Maltodextrin-binding proteins from diverse bacteria and archaea are potent solubility enhancers. FEBS letters. 2003;537(1-3):53-7.14. Fox JD, Routzahn KM, Bucher MH, Waugh DS. Maltodextrin-binding proteins from diverse bacteria and archaea are potent solubility enhancers. FEBS letters. 2003;537(1-3):53-7.
15. De Marco V, Stier G, Blandin S, de Marco A. The solubility and stability of recombinant proteins are increased by their fusion to NusA. Biochemical and biophysical research communications. 2004;322(3):766-71.15. De Marco V, Stier G, Blandin S, de Marco A. The solubility and stability of recombinant proteins are increased by their fusion to NusA. Biochemical and biophysical research communications. 2004;322(3):766-71.
16. Kuo D, Nie M, Courey AJ. SUMO as a solubility tag and in vivo cleavage of SUMO fusion proteins with Ulp1. Methods in molecular biology (Clifton, NJ). 2014;1177:71-80.16. Kuo D, Nie M, Courey AJ. SUMO as a solubility tag and in vivo cleavage of SUMO fusion proteins with Ulp1. Methods in molecular biology (Clifton, NJ). 2014;1177:71-80.
17. Peroutka Iii RJ, Orcutt SJ, Strickler JE, Butt TR. SUMO fusion technology for enhanced protein expression and purification in prokaryotes and eukaryotes. Methods in molecular biology (Clifton, NJ). 2011;705:15-30.17. Peroutka III RJ, Orcutt SJ, Strickler JE, Butt TR. SUMO fusion technology for enhanced protein expression and purification in prokaryotes and eukaryotes. Methods in molecular biology (Clifton, NJ). 2011;705:15-30.
18. Schafer F, Seip N, Maertens B, Block H, Kubicek J. Purification of GST-Tagged Proteins. Methods in enzymology. 2015;559:127-39.18. Schafer F, Seip N, Maertens B, Block H, Kubicek J. Purification of GST-Tagged Proteins. Methods in enzymes. 2015;559:127-39.
19. Kuhnel B, Alcantara J, Boothe J, van Rooijen G, Moloney M. Precise and efficient cleavage of recombinant fusion proteins using mammalian aspartic proteases. Protein engineering. 2003;16(10):777-83.19. Kuhnel B, Alcantara J, Boothe J, van Rooijen G, Moloney M. Precise and efficient cleavage of recombinant fusion proteins using mammalian aspartic proteases. Protein engineering. 2003;16(10):777-83.
20. Malakhov MP, Mattern MR, Malakhova OA, Drinker M, Weeks SD, Butt TR. SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins. Journal of structural and functional genomics. 2004;5(1-2):75-86.20. Malakhov MP, Matttern MR, Malakhova OA, Drinker M, Weeks SD, Butt TR. SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins. Journal of structural and functional genomics. 2004;5(1-2):75-86.
21. Choi SI, Han KS, Kim CW, Ryu KS, Kim BH, Kim KH, et al. Protein solubility and folding enhancement by interaction with RNA. PloS one. 2008;3(7):e2677.21. Choi SI, Han KS, Kim CW, Ryu KS, Kim BH, Kim KH, et al. Protein solubility and folding enhancement by interaction with RNA. PloS one. 2008;3(7):e2677.
22. Choi SI, Ryu K, Seong BL. RNA-mediated chaperone type for de novo protein folding. RNA biology. 2009;6(1):21-4.22. Choi SI, Ryu K, Seong BL. RNA-mediated chaperone type for de novo protein folding. RNA biology. 2009;6(1):21-4.
23. Son A, Choi SI, Han G, Seong BL. M1 RNA is important for the in-cell solubility of its cognate C5 protein: Implications for RNA-mediated protein folding. RNA biology. 2015;12(11):1198-208.23. Son A, Choi SI, Han G, Seong BL. M1 RNA is important for the in-cell solubility of its cognate C5 protein: Implications for RNA-mediated protein folding. RNA biology. 2015;12(11):1198-208.
24. Yang SW, Jang YH, Kwon SB, Lee YJ, Chae W, Byun YH, et al. Harnessing an RNA-mediated chaperone for the assembly of influenza hemagglutinin in an immunologically relevant conformation. FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 2018;32(5):2658-75.24. Yang SW, Jang YH, Kwon SB, Lee YJ, Chae W, Byun YH, et al. Harnessing an RNA-mediated chaperone for the assembly of influenza hemagglutinin in an immunologically relevant conformation. FASEB journal: official publication of the Federation of American Societies for Experimental Biology. 2018;32(5):2658-75.
25. Kim YS, Son A, Kim J, Kwon SB, Kim MH, Kim P, et al. Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles. Frontiers in immunology. 2018;9:1093.25. Kim YS, Son A, Kim J, Kwon SB, Kim MH, Kim P, et al. Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles. Frontiers in immunology. 2018;9:1093.
26. Lee KH, Seo SU, Song JM, Lee CM, Kim HA, Seong BL. Characterization of live influenza vaccine donor strain derived from cold-adaptation of X-31 virus. Vaccine. 2006;24(11):1966-74.26. Lee KH, Seo SU, Song JM, Lee CM, Kim HA, Seong BL. Characterization of live influenza vaccine donor strain derived from cold-adaptation of X-31 virus. Vaccine. 2006;24(11):1966-74.
27. Jang YH, Jung EJ, Lee KH, Byun YH, Yang SW, Seong BL. Genetic analysis of attenuation markers of cold-adapted X-31 influenza live vaccine donor strain. Vaccine. 2016;34(11):1343-9.27. Jang YH, Jung EJ, Lee KH, Byun YH, Yang SW, Seong BL. Genetic analysis of attenuation markers of cold-adapted X-31 influenza live vaccine donor strain. Vaccine. 2016;34(11):1343-9.
28. National Research Council Committee on Methods of Producing Monoclonal A. The National Academies Collection: Reports funded by National Institutes of Health. Monoclonal Antibody Production. Washington (DC): National Academies Press (US) National Academy of Sciences.; 1999.28. National Research Council Committee on Methods of Producing Monoclonal A. The National Academies Collection: Reports funded by National Institutes of Health. Monoclonal Antibody Production. Washington (DC): National Academies Press (US) National Academy of Sciences.; 1999.
29. Holzlohner P, Hanack K. Generation of Murine Monoclonal Antibodies by Hybridoma Technology. Journal of visualized experiments : JoVE. 2017(119).29. Holzlohner P, Hanack K. Generation of Murine Monoclonal Antibodies by Hybridoma Technology. Journal of visualized experiments: JoVE. 2017(119).
30. Du L, Kou Z, Ma C, Tao X, Wang L, Zhao G, et al. A truncated receptor-binding domain of MERS-CoV spike protein potently inhibits MERS-CoV infection and induces strong neutralizing antibody responses: implication for developing therapeutics and vaccines. PloS one. 2013;8(12):e81587.30. Du L, Kou Z, Ma C, Tao X, Wang L, Zhao G, et al. A truncated receptor-binding domain of MERS-CoV spike protein potently inhibits MERS-CoV infection and induces strong neutralizing antibody responses: implication for developing therapeutics and vaccines. PloS one. 2013;8(12):e81587.
31. Raj VS, Mou H, Smits SL, Dekkers DH, Muller MA, Dijkman R, et al. Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC. Nature. 2013;495(7440):251-4.31. Raj VS, Mou H, Smits SL, Dekkers DH, Muller MA, Dijkman R, et al. Dipeptidyl
<110> Industry-Academic Cooperation Foundation. Yonsei University <120> Method for producing target antigen-specific antibody using recombinant antigen <130> 1064587 <160> 11 <170> KoPatentIn 3.0 <210> 1 <211> 69 <212> PRT <213> Artificial Sequence <220> <223> mRID <400> 1 Met Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly Glu Gln Lys 1 5 10 15 Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Leu 20 25 30 Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu Asn 35 40 45 Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly Val Gly Ala 50 55 60 Glu Glu Glu Thr Leu 65 <210> 2 <211> 70 <212> PRT <213> Artificial Sequence <220> <223> rRID <400> 2 Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Glu Pro Lys 1 5 10 15 Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Val 20 25 30 Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu Ser 35 40 45 Gln Ala Thr Ala Ala Val Ala Asn His Ala Thr Asp Asn Gly Val Gly 50 55 60 Ala Glu Glu Glu Ser Leu 65 70 <210> 3 <211> 71 <212> PRT <213> Artificial Sequence <220> <223> hRID <400> 3 Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro 1 5 10 15 Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys 20 25 30 Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu 35 40 45 Ser Gln Ala Thr Ala Ala Ala Thr Asn His Thr Thr Asp Asn Gly Val 50 55 60 Gly Pro Glu Glu Glu Ser Val 65 70 <210> 4 <211> 186 <212> PRT <213> Artificial Sequence <220> <223> uHA stalk <400> 4 Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly Gly Trp Glu Gly 1 5 10 15 Leu Val Asp Gly Trp Tyr Gly Tyr His His Gln Asn Glu Gln Gly Val 20 25 30 Gly Tyr Ala Ala Asp Leu Lys Ser Thr Gln Asn Ala Ile Asp Lys Ile 35 40 45 Thr Asn Lys Val Asn Ile Val Ile Glu Lys Leu Asn Lys Gln Phe Glu 50 55 60 Ala Val Gly Lys Glu Phe Asn His Leu Glu Lys Arg Ile Glu Asn Leu 65 70 75 80 Asn Lys Lys Val Asp Asp Gly Phe Leu Asp Ile Trp Ala Tyr Asn Ala 85 90 95 Glu Leu Leu Val Leu Leu Glu Asn Glu Arg Thr Leu Asp Tyr His Asp 100 105 110 Ala Asn Val Lys Asn Leu Tyr Glu Lys Val Arg Leu Gln Leu Lys Asn 115 120 125 Asn Ala Lys Glu Ile Gly Asn Gly Cys Phe Glu Phe Tyr His Lys Cys 130 135 140 Asp Asn Glu Cys Leu Glu Ser Tyr Lys Asn Gly Thr Tyr Asp Tyr Pro 145 150 155 160 Lys Tyr Glu Glu Glu Ala Lys Leu Asn Arg Glu Glu Ile Asp Gly Val 165 170 175 Lys Leu Glu Leu Gly Val Tyr Gly Ile Leu 180 185 <210> 5 <211> 224 <212> PRT <213> Artificial Sequence <220> <223> HAgd <400> 5 Gly Ile Ala Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu 1 5 10 15 Leu Gly Asn Pro Glu Cys Asp Pro Leu Leu Pro Val Arg Ser Trp Ser 20 25 30 Tyr Ile Val Glu Thr Pro Asn Ser Glu Asn Gly Ile Cys Tyr Pro Gly 35 40 45 Asp Phe Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser 50 55 60 Ser Phe Glu Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn 65 70 75 80 His Asn Thr Asn Gly Val Thr Ala Ala Cys Ser His Glu Gly Lys Ser 85 90 95 Ser Phe Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr 100 105 110 Pro Lys Leu Lys Asn Ser Tyr Val Asn Lys Lys Gly Lys Glu Val Leu 115 120 125 Val Leu Trp Gly Ile His His Pro Pro Asn Ser Lys Glu Gln Gln Asn 130 135 140 Leu Tyr Gln Asn Glu Asn Ala Tyr Val Ser Val Val Thr Ser Asn Tyr 145 150 155 160 Asn Arg Arg Phe Thr Pro Glu Ile Ala Glu Arg Pro Lys Val Arg Asp 165 170 175 Gln Ala Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp 180 185 190 Thr Ile Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Met Tyr Ala 195 200 205 Phe Ala Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala 210 215 220 <210> 6 <211> 240 <212> PRT <213> Artificial Sequence <220> <223> MERS S1 RBD <400> 6 Glu Ala Lys Pro Ser Gly Ser Val Val Glu Gln Ala Glu Gly Val Glu 1 5 10 15 Cys Asp Phe Ser Pro Leu Leu Ser Gly Thr Pro Pro Gln Val Tyr Asn 20 25 30 Phe Lys Arg Leu Val Phe Thr Asn Cys Asn Tyr Asn Leu Thr Lys Leu 35 40 45 Leu Ser Leu Phe Ser Val Asn Asp Phe Thr Cys Ser Gln Ile Ser Pro 50 55 60 Ala Ala Ile Ala Ser Asn Cys Tyr Ser Ser Leu Ile Leu Asp Tyr Phe 65 70 75 80 Ser Tyr Pro Leu Ser Met Lys Ser Asp Leu Ser Val Ser Ser Ala Gly 85 90 95 Pro Ile Ser Gln Phe Asn Tyr Lys Gln Ser Phe Ser Asn Pro Thr Cys 100 105 110 Leu Ile Leu Ala Thr Val Pro His Asn Leu Thr Thr Ile Thr Lys Pro 115 120 125 Leu Lys Tyr Ser Tyr Ile Asn Lys Cys Ser Arg Leu Leu Ser Asp Asp 130 135 140 Arg Thr Glu Val Pro Gln Leu Val Asn Ala Asn Gln Tyr Ser Pro Cys 145 150 155 160 Val Ser Ile Val Pro Ser Thr Val Trp Glu Asp Gly Asp Tyr Tyr Arg 165 170 175 Lys Gln Leu Ser Pro Leu Glu Gly Gly Gly Trp Leu Val Ala Ser Gly 180 185 190 Ser Thr Val Ala Met Thr Glu Gln Leu Gln Met Gly Phe Gly Ile Thr 195 200 205 Val Gln Tyr Gly Thr Asp Thr Asn Ser Val Cys Pro Lys Leu Glu Phe 210 215 220 Ala Asn Asp Thr Lys Ile Ala Ser Gln Leu Gly Asn Cys Val Glu Tyr 225 230 235 240 <210> 7 <211> 282 <212> PRT <213> Artificial Sequence <220> <223> mRID-uHAstalk <400> 7 Met Ser Glu Gln Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly 1 5 10 15 Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Asn Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly 50 55 60 Val Gly Ala Glu Glu Glu Thr Leu Asp Asp Asp Asp Asp Asp Ser Gly 65 70 75 80 Glu Asn Leu Tyr Phe Gln Gly Thr Gly Leu Phe Gly Ala Ile Ala Gly 85 90 95 Phe Ile Glu Gly Gly Trp Glu Gly Leu Val Asp Gly Trp Tyr Gly Tyr 100 105 110 His His Gln Asn Glu Gln Gly Val Gly Tyr Ala Ala Asp Leu Lys Ser 115 120 125 Thr Gln Asn Ala Ile Asp Lys Ile Thr Asn Lys Val Asn Ile Val Ile 130 135 140 Glu Lys Leu Asn Lys Gln Phe Glu Ala Val Gly Lys Glu Phe Asn His 145 150 155 160 Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 165 170 175 Leu Asp Ile Trp Ala Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 180 185 190 Glu Arg Thr Leu Asp Tyr His Asp Ala Asn Val Lys Asn Leu Tyr Glu 195 200 205 Lys Val Arg Leu Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly 210 215 220 Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Glu Cys Leu Glu Ser Tyr 225 230 235 240 Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Glu Glu Glu Ala Lys Leu 245 250 255 Asn Arg Glu Glu Ile Asp Gly Val Lys Leu Glu Leu Gly Val Tyr Gly 260 265 270 Ile Leu Lys Leu His His His His His His 275 280 <210> 8 <211> 321 <212> PRT <213> Artificial Sequence <220> <223> mRID-HAgd <400> 8 Met Ser Glu Gln Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly 1 5 10 15 Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Asn Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly 50 55 60 Val Gly Ala Glu Glu Glu Thr Leu Asp Asp Asp Asp Asp Asp Ser Gly 65 70 75 80 Glu Asn Leu Tyr Phe Gln Gly Thr Gly Ile Ala Pro Leu Gln Leu Gly 85 90 95 Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly Asn Pro Glu Cys Asp Pro 100 105 110 Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile Val Glu Thr Pro Asn Ser 115 120 125 Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe Ile Asp Tyr Glu Glu Leu 130 135 140 Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu Arg Phe Glu Ile Phe 145 150 155 160 Pro Lys Glu Ser Ser Trp Pro Asn His Asn Thr Thr Lys Gly Val Thr 165 170 175 Ala Ala Cys Ser His Ala Gly Lys Ser Ser Phe Tyr Arg Asn Leu Leu 180 185 190 Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys Leu Lys Asn Ser Tyr 195 200 205 Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu Trp Gly Ile His His 210 215 220 Pro Ser Asn Ser Lys Asp Gln Gln Asn Ile Tyr Gln Asn Glu Asn Ala 225 230 235 240 Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg Arg Phe Thr Pro Glu 245 250 255 Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala Gly Arg Met Asn Tyr 260 265 270 Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile Ile Phe Glu Ala Asn 275 280 285 Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala Leu Ser Arg Gly Phe 290 295 300 Gly Ser Gly Ile Ile Thr Ser Asn Ala Lys Leu His His His His His 305 310 315 320 His <210> 9 <211> 322 <212> PRT <213> Artificial Sequence <220> <223> rRID-HAgd <400> 9 Met Ser Glu Gln Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly 1 5 10 15 Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Ser Gln Ala Thr Ala Ala Val Ala Asn His Ala Thr Asp Asn 50 55 60 Gly Val Gly Ala Glu Glu Glu Ser Leu Asp Asp Asp Asp Asp Asp Ser 65 70 75 80 Gly Glu Asn Leu Tyr Phe Gln Gly Thr Gly Ile Ala Pro Leu Gln Leu 85 90 95 Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly Asn Pro Glu Cys Asp 100 105 110 Pro Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile Val Glu Thr Pro Asn 115 120 125 Ser Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe Ile Asp Tyr Glu Glu 130 135 140 Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu Arg Phe Glu Ile 145 150 155 160 Phe Pro Lys Glu Ser Ser Trp Pro Asn His Asn Thr Thr Lys Gly Val 165 170 175 Thr Ala Ala Cys Ser His Ala Gly Lys Ser Ser Phe Tyr Arg Asn Leu 180 185 190 Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys Leu Lys Asn Ser 195 200 205 Tyr Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu Trp Gly Ile His 210 215 220 His Pro Ser Asn Ser Lys Asp Gln Gln Asn Ile Tyr Gln Asn Glu Asn 225 230 235 240 Ala Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg Arg Phe Thr Pro 245 250 255 Glu Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala Gly Arg Met Asn 260 265 270 Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile Ile Phe Glu Ala 275 280 285 Asn Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala Leu Ser Arg Gly 290 295 300 Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala Lys Leu His His His His 305 310 315 320 His His <210> 10 <211> 326 <212> PRT <213> Artificial Sequence <220> <223> hRID-HAgd <400> 10 Met Ser Glu Gln His Ala Gln Ala Ala Val Gln Ala Ala Glu Val Lys 1 5 10 15 Val Asp Gly Ser Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg 20 25 30 Leu Lys Ala Glu Lys Lys Val Ala Glu Lys Glu Ala Lys Gln Lys Glu 35 40 45 Leu Ser Glu Lys Gln Leu Ser Gln Ala Thr Ala Ala Ala Thr Asn His 50 55 60 Thr Thr Asp Asn Gly Val Gly Pro Glu Glu Glu Ser Val Asp Asp Asp 65 70 75 80 Asp Asp Asp Ser Gly Glu Asn Leu Tyr Phe Gln Gly Thr Gly Ile Ala 85 90 95 Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly Asn 100 105 110 Pro Glu Cys Asp Pro Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile Val 115 120 125 Glu Thr Pro Asn Ser Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe Ile 130 135 140 Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu 145 150 155 160 Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn His Asn Thr 165 170 175 Thr Lys Gly Val Thr Ala Ala Cys Ser His Ala Gly Lys Ser Ser Phe 180 185 190 Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys 195 200 205 Leu Lys Asn Ser Tyr Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu 210 215 220 Trp Gly Ile His His Pro Ser Asn Ser Lys Asp Gln Gln Asn Ile Tyr 225 230 235 240 Gln Asn Glu Asn Ala Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg 245 250 255 Arg Phe Thr Pro Glu Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala 260 265 270 Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile 275 280 285 Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala 290 295 300 Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala Lys Leu 305 310 315 320 His His His His His His 325 <210> 11 <211> 338 <212> PRT <213> Artificial Sequence <220> <223> mRID-MERS S1 RBD <400> 11 Met Ser Glu Gln Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly 1 5 10 15 Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Asn Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly 50 55 60 Val Gly Ala Glu Glu Glu Thr Leu Asp Asp Asp Asp Asp Asp Ser Gly 65 70 75 80 Glu Asn Leu Tyr Phe Gln Gly Thr Glu Ala Lys Pro Ser Gly Ser Val 85 90 95 Val Glu Gln Ala Glu Gly Val Glu Cys Asp Phe Ser Pro Leu Leu Ser 100 105 110 Gly Thr Pro Pro Gln Val Tyr Asn Phe Lys Arg Leu Val Phe Thr Asn 115 120 125 Cys Asn Tyr Asn Leu Thr Lys Leu Leu Ser Leu Phe Ser Val Asn Asp 130 135 140 Phe Thr Cys Ser Gln Ile Ser Pro Ala Ala Ile Ala Ser Asn Cys Tyr 145 150 155 160 Ser Ser Leu Ile Leu Asp Tyr Phe Ser Tyr Pro Leu Ser Met Lys Ser 165 170 175 Asp Leu Ser Val Ser Ser Ala Gly Pro Ile Ser Gln Phe Asn Tyr Lys 180 185 190 Gln Ser Phe Ser Asn Pro Thr Cys Leu Ile Leu Ala Thr Val Pro His 195 200 205 Asn Leu Thr Thr Ile Thr Lys Pro Leu Lys Tyr Ser Tyr Ile Asn Lys 210 215 220 Cys Ser Arg Leu Leu Ser Asp Asp Arg Thr Glu Val Pro Gln Leu Val 225 230 235 240 Asn Ala Asn Gln Tyr Ser Pro Cys Val Ser Ile Val Pro Ser Thr Val 245 250 255 Trp Glu Asp Gly Asp Tyr Tyr Arg Lys Gln Leu Ser Pro Leu Glu Gly 260 265 270 Gly Gly Trp Leu Val Ala Ser Gly Ser Thr Val Ala Met Thr Glu Gln 275 280 285 Leu Gln Met Gly Phe Gly Ile Thr Val Gln Tyr Gly Thr Asp Thr Asn 290 295 300 Ser Val Cys Pro Lys Leu Glu Phe Ala Asn Asp Thr Lys Ile Ala Ser 305 310 315 320 Gln Leu Gly Asn Cys Val Glu Tyr Val Asp Lys Leu His His His His 325 330 335 His His <110> Industry-Academic Cooperation Foundation. Yonsei University <120> Method for producing target antigen-specific antibody using recombinant antigen <130> 1064587 <160> 11 <170> KoPatentIn 3.0 <210> 1 <211> 69 <212> PRT <213> Artificial Sequence <220> <223> mRID <400> 1 Met Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly Glu Gln Lys 1 5 10 15 Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Leu 20 25 30 Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu Asn 35 40 45 Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly Val Gly Ala 50 55 60 Glu Glu Glu Thr Leu 65 <210> 2 <211> 70 <212> PRT <213> Artificial Sequence <220> <223> rRID <400> 2 Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Glu Pro Lys 1 5 10 15 Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Val 20 25 30 Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu Ser 35 40 45 Gln Ala Thr Ala Ala Val Ala Asn His Ala Thr Asp Asn Gly Val Gly 50 55 60 Ala Glu Glu Glu Ser Leu 65 70 <210> 3 <211> 71 <212> PRT <213> Artificial Sequence <220> <223> hRID <400> 3 Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro 1 5 10 15 Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys 20 25 30 Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu 35 40 45 Ser Gln Ala Thr Ala Ala Ala Thr Asn His Thr Thr Asp Asn Gly Val 50 55 60 Gly Pro Glu Glu Glu Ser Val 65 70 <210> 4 <211> 186 <212> PRT <213> Artificial Sequence <220> <223> uHA stalk <400> 4 Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly Gly Trp Glu Gly 1 5 10 15 Leu Val Asp Gly Trp Tyr Gly Tyr His His Gln Asn Glu Gln Gly Val 20 25 30 Gly Tyr Ala Ala Asp Leu Lys Ser Thr Gln Asn Ala Ile Asp Lys Ile 35 40 45 Thr Asn Lys Val Asn Ile Val Ile Glu Lys Leu Asn Lys Gln Phe Glu 50 55 60 Ala Val Gly Lys Glu Phe Asn His Leu Glu Lys Arg Ile Glu Asn Leu 65 70 75 80 Asn Lys Lys Val Asp Asp Gly Phe Leu Asp Ile Trp Ala Tyr Asn Ala 85 90 95 Glu Leu Leu Val Leu Leu Glu Asn Glu Arg Thr Leu Asp Tyr His Asp 100 105 110 Ala Asn Val Lys Asn Leu Tyr Glu Lys Val Arg Leu Gln Leu Lys Asn 115 120 125 Asn Ala Lys Glu Ile Gly Asn Gly Cys Phe Glu Phe Tyr His Lys Cys 130 135 140 Asp Asn Glu Cys Leu Glu Ser Tyr Lys Asn Gly Thr Tyr Asp Tyr Pro 145 150 155 160 Lys Tyr Glu Glu Glu Ala Lys Leu Asn Arg Glu Glu Ile Asp Gly Val 165 170 175 Lys Leu Glu Leu Gly Val Tyr Gly Ile Leu 180 185 <210> 5 <211> 224 <212> PRT <213> Artificial Sequence <220> <223> HAgd <400> 5 Gly Ile Ala Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu 1 5 10 15 Leu Gly Asn Pro Glu Cys Asp Pro Leu Leu Pro Val Arg Ser Trp Ser 20 25 30 Tyr Ile Val Glu Thr Pro Asn Ser Glu Asn Gly Ile Cys Tyr Pro Gly 35 40 45 Asp Phe Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser 50 55 60 Ser Phe Glu Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn 65 70 75 80 His Asn Thr Asn Gly Val Thr Ala Ala Cys Ser His Glu Gly Lys Ser 85 90 95 Ser Phe Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr 100 105 110 Pro Lys Leu Lys Asn Ser Tyr Val Asn Lys Lys Gly Lys Glu Val Leu 115 120 125 Val Leu Trp Gly Ile His His Pro Asn Ser Lys Glu Gln Gln Asn 130 135 140 Leu Tyr Gln Asn Glu Asn Ala Tyr Val Ser Val Val Thr Ser Asn Tyr 145 150 155 160 Asn Arg Arg Phe Thr Pro Glu Ile Ala Glu Arg Pro Lys Val Arg Asp 165 170 175 Gln Ala Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp 180 185 190 Thr Ile Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Met Tyr Ala 195 200 205 Phe Ala Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala 210 215 220 <210> 6 <211> 240 <212> PRT <213> Artificial Sequence <220> <223> MERS S1 RBD <400> 6 Glu Ala Lys Pro Ser Gly Ser Val Val Glu Gln Ala Glu Gly Val Glu 1 5 10 15 Cys Asp Phe Ser Pro Leu Leu Ser Gly Thr Pro Pro Gln Val Tyr Asn 20 25 30 Phe Lys Arg Leu Val Phe Thr Asn Cys Asn Tyr Asn Leu Thr Lys Leu 35 40 45 Leu Ser Leu Phe Ser Val Asn Asp Phe Thr Cys Ser Gln Ile Ser Pro 50 55 60 Ala Ala Ile Ala Ser Asn Cys Tyr Ser Ser Leu Ile Leu Asp Tyr Phe 65 70 75 80 Ser Tyr Pro Leu Ser Met Lys Ser Asp Leu Ser Val Ser Ser Ala Gly 85 90 95 Pro Ile Ser Gln Phe Asn Tyr Lys Gln Ser Phe Ser Asn Pro Thr Cys 100 105 110 Leu Ile Leu Ala Thr Val Pro His Asn Leu Thr Thr Ile Thr Lys Pro 115 120 125 Leu Lys Tyr Ser Tyr Ile Asn Lys Cys Ser Arg Leu Leu Ser Asp Asp 130 135 140 Arg Thr Glu Val Pro Gln Leu Val Asn Ala Asn Gln Tyr Ser Pro Cys 145 150 155 160 Val Ser Ile Val Pro Ser Thr Val Trp Glu Asp Gly Asp Tyr Tyr Arg 165 170 175 Lys Gln Leu Ser Pro Leu Glu Gly Gly Gly Trp Leu Val Ala Ser Gly 180 185 190 Ser Thr Val Ala Met Thr Glu Gln Leu Gln Met Gly Phe Gly Ile Thr 195 200 205 Val Gln Tyr Gly Thr Asp Thr Asn Ser Val Cys Pro Lys Leu Glu Phe 210 215 220 Ala Asn Asp Thr Lys Ile Ala Ser Gln Leu Gly Asn Cys Val Glu Tyr 225 230 235 240 <210> 7 <211> 282 <212> PRT <213> Artificial Sequence <220> <223> mRID-uHAstalk <400> 7 Met Ser Glu Gln Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly 1 5 10 15 Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Asn Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly 50 55 60 Val Gly Ala Glu Glu Glu Thr Leu Asp Asp Asp Asp Asp Asp Asp Ser Gly 65 70 75 80 Glu Asn Leu Tyr Phe Gln Gly Thr Gly Leu Phe Gly Ala Ile Ala Gly 85 90 95 Phe Ile Glu Gly Gly Trp Glu Gly Leu Val Asp Gly Trp Tyr Gly Tyr 100 105 110 His His Gln Asn Glu Gln Gly Val Gly Tyr Ala Ala Asp Leu Lys Ser 115 120 125 Thr Gln Asn Ala Ile Asp Lys Ile Thr Asn Lys Val Asn Ile Val Ile 130 135 140 Glu Lys Leu Asn Lys Gln Phe Glu Ala Val Gly Lys Glu Phe Asn His 145 150 155 160 Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 165 170 175 Leu Asp Ile Trp Ala Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 180 185 190 Glu Arg Thr Leu Asp Tyr His Asp Ala Asn Val Lys Asn Leu Tyr Glu 195 200 205 Lys Val Arg Leu Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly 210 215 220 Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Glu Cys Leu Glu Ser Tyr 225 230 235 240 Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Glu Glu Glu Ala Lys Leu 245 250 255 Asn Arg Glu Glu Ile Asp Gly Val Lys Leu Glu Leu Gly Val Tyr Gly 260 265 270 Ile Leu Lys Leu His His His His His His 275 280 <210> 8 <211> 321 <212> PRT <213> Artificial Sequence <220> <223> mRID-HAgd <400> 8 Met Ser Glu Gln Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly 1 5 10 15 Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Asn Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly 50 55 60 Val Gly Ala Glu Glu Glu Thr Leu Asp Asp Asp Asp Asp Asp Asp Ser Gly 65 70 75 80 Glu Asn Leu Tyr Phe Gln Gly Thr Gly Ile Ala Pro Leu Gln Leu Gly 85 90 95 Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly Asn Pro Glu Cys Asp Pro 100 105 110 Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile Val Glu Thr Pro Asn Ser 115 120 125 Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe Ile Asp Tyr Glu Glu Leu 130 135 140 Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu Arg Phe Glu Ile Phe 145 150 155 160 Pro Lys Glu Ser Ser Trp Pro Asn His Asn Thr Thr Lys Gly Val Thr 165 170 175 Ala Ala Cys Ser His Ala Gly Lys Ser Ser Phe Tyr Arg Asn Leu Leu 180 185 190 Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys Leu Lys Asn Ser Tyr 195 200 205 Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu Trp Gly Ile His His 210 215 220 Pro Ser Asn Ser Lys Asp Gln Gln Asn Ile Tyr Gln Asn Glu Asn Ala 225 230 235 240 Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg Arg Phe Thr Pro Glu 245 250 255 Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala Gly Arg Met Asn Tyr 260 265 270 Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile Ile Phe Glu Ala Asn 275 280 285 Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala Leu Ser Arg Gly Phe 290 295 300 Gly Ser Gly Ile Ile Thr Ser Asn Ala Lys Leu His His His His His 305 310 315 320 His <210> 9 <211> 322 <212> PRT <213> Artificial Sequence <220> <223> rRID-HAgd <400> 9 Met Ser Glu Gln Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly 1 5 10 15 Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Ser Gln Ala Thr Ala Ala Val Ala Asn His Ala Thr Asp Asn 50 55 60 Gly Val Gly Ala Glu Glu Glu Ser Leu Asp Asp Asp Asp Asp Asp Ser 65 70 75 80 Gly Glu Asn Leu Tyr Phe Gln Gly Thr Gly Ile Ala Pro Leu Gln Leu 85 90 95 Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly Asn Pro Glu Cys Asp 100 105 110 Pro Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile Val Glu Thr Pro Asn 115 120 125 Ser Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe Ile Asp Tyr Glu Glu 130 135 140 Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu Arg Phe Glu Ile 145 150 155 160 Phe Pro Lys Glu Ser Ser Trp Pro Asn His Asn Thr Thr Lys Gly Val 165 170 175 Thr Ala Ala Cys Ser His Ala Gly Lys Ser Ser Phe Tyr Arg Asn Leu 180 185 190 Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys Leu Lys Asn Ser 195 200 205 Tyr Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu Trp Gly Ile His 210 215 220 His Pro Ser Asn Ser Lys Asp Gln Gln Asn Ile Tyr Gln Asn Glu Asn 225 230 235 240 Ala Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg Arg Phe Thr Pro 245 250 255 Glu Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala Gly Arg Met Asn 260 265 270 Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile Ile Phe Glu Ala 275 280 285 Asn Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala Leu Ser Arg Gly 290 295 300 Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala Lys Leu His His His His 305 310 315 320 His His <210> 10 <211> 326 <212> PRT <213> Artificial Sequence <220> <223> hRID-HAgd <400> 10 Met Ser Glu Gln His Ala Gln Ala Ala Val Gln Ala Ala Glu Val Lys 1 5 10 15 Val Asp Gly Ser Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg 20 25 30 Leu Lys Ala Glu Lys Lys Val Ala Glu Lys Glu Ala Lys Gln Lys Glu 35 40 45 Leu Ser Glu Lys Gln Leu Ser Gln Ala Thr Ala Ala Ala Thr Asn His 50 55 60 Thr Thr Asp Asn Gly Val Gly Pro Glu Glu Glu Ser Val Asp Asp Asp 65 70 75 80 Asp Asp Asp Ser Gly Glu Asn Leu Tyr Phe Gln Gly Thr Gly Ile Ala 85 90 95 Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly Asn 100 105 110 Pro Glu Cys Asp Pro Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile Val 115 120 125 Glu Thr Pro Asn Ser Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe Ile 130 135 140 Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu 145 150 155 160 Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn His Asn Thr 165 170 175 Thr Lys Gly Val Thr Ala Ala Cys Ser His Ala Gly Lys Ser Ser Phe 180 185 190 Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys 195 200 205 Leu Lys Asn Ser Tyr Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu 210 215 220 Trp Gly Ile His His Pro Ser Asn Ser Lys Asp Gln Gln Asn Ile Tyr 225 230 235 240 Gln Asn Glu Asn Ala Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg 245 250 255 Arg Phe Thr Pro Glu Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala 260 265 270 Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile 275 280 285 Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala 290 295 300 Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala Lys Leu 305 310 315 320 His His His His His His 325 <210> 11 <211> 338 <212> PRT <213> Artificial Sequence <220> <223> mRID-MERS S1 RBD <400> 11 Met Ser Glu Gln Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly 1 5 10 15 Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu 20 25 30 Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys 35 40 45 Gln Leu Asn Gln Thr Ala Ser Ala Pro Asn His Thr Ala Asp Asn Gly 50 55 60 Val Gly Ala Glu Glu Glu Thr Leu Asp Asp Asp Asp Asp Asp Asp Ser Gly 65 70 75 80 Glu Asn Leu Tyr Phe Gln Gly Thr Glu Ala Lys Pro Ser Gly Ser Val 85 90 95 Val Glu Gln Ala Glu Gly Val Glu Cys Asp Phe Ser Pro Leu Leu Ser 100 105 110 Gly Thr Pro Pro Gln Val Tyr Asn Phe Lys Arg Leu Val Phe Thr Asn 115 120 125 Cys Asn Tyr Asn Leu Thr Lys Leu Leu Ser Leu Phe Ser Val Asn Asp 130 135 140 Phe Thr Cys Ser Gln Ile Ser Pro Ala Ala Ile Ala Ser Asn Cys Tyr 145 150 155 160 Ser Ser Leu Ile Leu Asp Tyr Phe Ser Tyr Pro Leu Ser Met Lys Ser 165 170 175 Asp Leu Ser Val Ser Ser Ala Gly Pro Ile Ser Gln Phe Asn Tyr Lys 180 185 190 Gln Ser Phe Ser Asn Pro Thr Cys Leu Ile Leu Ala Thr Val Pro His 195 200 205 Asn Leu Thr Thr Ile Thr Lys Pro Leu Lys Tyr Ser Tyr Ile Asn Lys 210 215 220 Cys Ser Arg Leu Leu Ser Asp Asp Arg Thr Glu Val Pro Gln Leu Val 225 230 235 240 Asn Ala Asn Gln Tyr Ser Pro Cys Val Ser Ile Val Pro Ser Thr Val 245 250 255 Trp Glu Asp Gly Asp Tyr Tyr Arg Lys Gln Leu Ser Pro Leu Glu Gly 260 265 270 Gly Gly Trp Leu Val Ala Ser Gly Ser Thr Val Ala Met Thr Glu Gln 275 280 285 Leu Gln Met Gly Phe Gly Ile Thr Val Gln Tyr Gly Thr Asp Thr Asn 290 295 300 Ser Val Cys Pro Lys Leu Glu Phe Ala Asn Asp Thr Lys Ile Ala Ser 305 310 315 320 Gln Leu Gly Asn Cys Val Glu Tyr Val Asp Lys Leu His His His His 325 330 335 His His
Claims (10)
상기 융합파트너는 상기 숙주동물과 동일한 포유동물로부터 유래된 자가 항원(self-antigen)이고, 상기 포유동물로부터 유래된 아미노아실 tRNA 합성효소에서 분리한 RNA 결합 도메인(Aminoacyl tRNA synthetase appended RNA interacting domain; RID)의 아미노산 서열을 포함하며,
상기 목적 항원은 이종 항원(foreign-antigen)인 것인, 목적 항원 특이적 항체 제조방법.
a fusion partner for enhancing the expression efficiency of the target antigen; and administering to a host animal a recombinant protein comprising the target antigen represented by the amino acid sequence of SEQ ID NO: 6,
The fusion partner is a self-antigen derived from the same mammal as the host animal, and an RNA binding domain (Aminoacyl tRNA synthetase appended RNA interacting domain; RID) isolated from an aminoacyl tRNA synthetase derived from the mammal. ) comprising the amino acid sequence of
The target antigen is a foreign antigen (foreign-antigen), the target antigen-specific antibody production method.
상기 RID는 서열번호 1, 서열번호 2 및 서열번호 3으로 이루어진 군에서 선택되는 어느 하나의 아미노산 서열로 표시되는 것인, 목적 항원 특이적 항체 제조방법.
The method of claim 1,
Wherein the RID is represented by any one amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, the method for producing an antigen-specific antibody of interest.
상기 숙주동물은 마우스, 토끼, 말, 개, 고양이, 닭 및 오리로 이루어진 군에서 선택되는 어느 하나 이상인 것인, 목적 항원 특이적 항체 제조방법.
The method of claim 1,
The host animal is any one or more selected from the group consisting of mice, rabbits, horses, dogs, cats, chickens and ducks, the method for producing an antigen-specific antibody of interest.
A target antigen-specific antibody prepared by the method according to claim 1 and specifically recognizing the peptide represented by the amino acid sequence consisting of SEQ ID NO: 6.
상기 목적 항원 특이적 항체는 다클론 항체 또는 단일클론 항체인, 목적 항원 특이적 항체.
The method of claim 5,
The antigen-specific antibody of interest is a polyclonal antibody or a monoclonal antibody, an antigen-specific antibody of interest.
A composition for diagnosing viral infection comprising the antigen-specific antibody of claim 5 .
상기 목적 항원 특이적 항체와 샘플의 반응을 검출하는 단계를 포함하는, 대상체의 바이러스 감염 여부에 대한 정보제공 방법.Contacting the sample with the antigen-specific antibody of claim 5; and
A method for providing information on whether a subject is infected with a virus, comprising the step of detecting a reaction between the target antigen-specific antibody and the sample.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020180137394A KR102259974B1 (en) | 2018-11-09 | 2018-11-09 | Method for producing target antigen-specific antibody using recombinant antigen |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020180137394A KR102259974B1 (en) | 2018-11-09 | 2018-11-09 | Method for producing target antigen-specific antibody using recombinant antigen |
Publications (2)
Publication Number | Publication Date |
---|---|
KR20200053906A KR20200053906A (en) | 2020-05-19 |
KR102259974B1 true KR102259974B1 (en) | 2021-06-01 |
Family
ID=70913359
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020180137394A KR102259974B1 (en) | 2018-11-09 | 2018-11-09 | Method for producing target antigen-specific antibody using recombinant antigen |
Country Status (1)
Country | Link |
---|---|
KR (1) | KR102259974B1 (en) |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101908438B1 (en) * | 2016-02-05 | 2018-10-16 | (주)피앤피바이오팜 | A MANUFACTURING METHOD FOR SOLUBLE AND ACTIVE RECOMBINANT PROTEIN USING Aminoacyl tRNA synthetase N-terminal domain AS FUSION PARTNER AND A PRODUCT THEREOF |
KR101914779B1 (en) * | 2016-03-18 | 2018-11-02 | (주)인테라 | Recombinant Expression Vectors for Producing Norovirus Vaccine |
KR102018602B1 (en) * | 2016-08-22 | 2019-09-09 | (주)인테라 | A MANUFACTURING METHOD FOR SOLUBLE AND ACTIVE RECOMBINANT PROTEIN USING Aminoacyl tRNA synthetase N-terminal domain AS FUSION PARTNER AND A PRODUCT THEREOF |
-
2018
- 2018-11-09 KR KR1020180137394A patent/KR102259974B1/en active IP Right Grant
Non-Patent Citations (1)
Title |
---|
Int. J. Mol. Sci., Vol. 19, pp. 3016(2018.10.03.) |
Also Published As
Publication number | Publication date |
---|---|
KR20200053906A (en) | 2020-05-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11479605B2 (en) | Homodimeric protein constructs | |
CN109796531B (en) | Monoclonal antibody of swine Delta coronavirus N protein, epitope and application thereof | |
KR101914779B1 (en) | Recombinant Expression Vectors for Producing Norovirus Vaccine | |
Ogun et al. | The oligomerization domain of C4-binding protein (C4bp) acts as an adjuvant, and the fusion protein comprised of the 19-kilodalton merozoite surface protein 1 fused with the murine C4bp domain protects mice against malaria | |
EA035375B1 (en) | Influenza virus vaccines and uses thereof | |
CN109627294B (en) | Correctly folded recombinant rabies virus G protein extracellular domain and potential application thereof | |
CN112920278B (en) | Novel coronavirus specific fusion protein antigen and preparation method and application thereof | |
US20140234316A1 (en) | Vaccibodies targeted to cross-presenting dendritic cells | |
CN113018427A (en) | Multivalent fusion protein vaccine based on neutralizing epitope of new coronavirus | |
Klausberger et al. | Globular head-displayed conserved influenza H1 hemagglutinin stalk epitopes confer protection against heterologous H1N1 virus | |
KR20180087431A (en) | FKBP domain having a transglutaminase recognition site | |
JP2020503839A (en) | A novel human immunodeficiency virus envelope protein antigen expressed in mammals | |
CN113527475B (en) | Hybridoma cell secreting novel duck reovirus sigma C protein monoclonal antibody, monoclonal antibody and application | |
KR101972899B1 (en) | Fusion protein comprising human C4bp oligomerization domain linked to hagfish VLRB protein with deleted hydrophobic tail domain and uses thereof | |
CN105753980B (en) | A kind of HPV18 E6 monoclonal antibody and its preparation method and application | |
CN110003314B (en) | Epitope peptide capable of inducing broad-spectrum protective antibody by H1N1 influenza virus hemagglutinin and application thereof | |
Kwon et al. | Recombinant adenylate kinase 3 from liver fluke Clonorchis sinensis for histochemical analysis and serodiagnosis of clonorchiasis | |
KR102259974B1 (en) | Method for producing target antigen-specific antibody using recombinant antigen | |
Yu et al. | Fine mapping and conservation analysis of linear B-cell epitopes of peste des petits ruminants virus hemagglutinin protein | |
CN109400684B (en) | PEDV S-RBD linear B cell epitope, two specific recognition monoclonal antibodies and application | |
CN114891075B (en) | Polypeptide with binding affinity to novel coronavirus S protein RBMFP structural domain and application thereof | |
EP2154251B1 (en) | Expression system of recombinant proteins with enhanced yield and immunogenicity | |
WO2022038501A1 (en) | Fusion proteins comprising sars-cov-2 receptor binding domain | |
CN113736825A (en) | Recombinant drosophila cell line for expressing swine atypical pestivirus fusion protein and preparation method and application thereof | |
CN109504667B (en) | IRAK-M polyclonal antibody and preparation method thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AMND | Amendment | ||
E601 | Decision to refuse application | ||
AMND | Amendment | ||
E902 | Notification of reason for refusal | ||
X701 | Decision to grant (after re-examination) | ||
GRNT | Written decision to grant |