CN115043934B - Nanometer antibody targeting novel coronavirus and preparation method and application thereof - Google Patents
Nanometer antibody targeting novel coronavirus and preparation method and application thereof Download PDFInfo
- Publication number
- CN115043934B CN115043934B CN202210637521.2A CN202210637521A CN115043934B CN 115043934 B CN115043934 B CN 115043934B CN 202210637521 A CN202210637521 A CN 202210637521A CN 115043934 B CN115043934 B CN 115043934B
- Authority
- CN
- China
- Prior art keywords
- nanobody
- gly
- novel coronavirus
- ser
- seq
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 93
- 230000008685 targeting Effects 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 238000001514 detection method Methods 0.000 claims description 54
- 206010035664 Pneumonia Diseases 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 8
- 230000027455 binding Effects 0.000 abstract description 25
- 241000700605 Viruses Species 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 12
- 101100112922 Candida albicans CDR3 gene Proteins 0.000 abstract description 11
- 230000009286 beneficial effect Effects 0.000 abstract description 8
- 230000001105 regulatory effect Effects 0.000 abstract description 4
- 108091005634 SARS-CoV-2 receptor-binding domains Proteins 0.000 description 83
- 150000001413 amino acids Chemical class 0.000 description 62
- 108090000623 proteins and genes Proteins 0.000 description 37
- 102000004169 proteins and genes Human genes 0.000 description 28
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 25
- 238000012216 screening Methods 0.000 description 21
- 241001678559 COVID-19 virus Species 0.000 description 16
- 238000004458 analytical method Methods 0.000 description 16
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 14
- 238000002965 ELISA Methods 0.000 description 12
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 12
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 12
- 108090000975 Angiotensin-converting enzyme 2 Proteins 0.000 description 11
- 238000012163 sequencing technique Methods 0.000 description 11
- 102100035765 Angiotensin-converting enzyme 2 Human genes 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- YNQIDCRRTWGHJD-ZLUOBGJFSA-N Asp-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(O)=O YNQIDCRRTWGHJD-ZLUOBGJFSA-N 0.000 description 9
- SBCYJMOOHUDWDA-NUMRIWBASA-N Glu-Asp-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SBCYJMOOHUDWDA-NUMRIWBASA-N 0.000 description 9
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 9
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 9
- 238000010586 diagram Methods 0.000 description 9
- 239000013612 plasmid Substances 0.000 description 9
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 8
- 230000003472 neutralizing effect Effects 0.000 description 8
- 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 7
- 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 7
- QUCDKEKDPYISNX-HJGDQZAQSA-N Lys-Asn-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QUCDKEKDPYISNX-HJGDQZAQSA-N 0.000 description 7
- YMTLKLXDFCSCNX-BYPYZUCNSA-N Ser-Gly-Gly Chemical compound OC[C@H](N)C(=O)NCC(=O)NCC(O)=O YMTLKLXDFCSCNX-BYPYZUCNSA-N 0.000 description 7
- ZCUFMRIQCPNOHZ-NRPADANISA-N Ala-Val-Gln Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N ZCUFMRIQCPNOHZ-NRPADANISA-N 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 6
- IOFDDSNZJDIGPB-GVXVVHGQSA-N Gln-Leu-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IOFDDSNZJDIGPB-GVXVVHGQSA-N 0.000 description 6
- RFTVTKBHDXCEEX-WDSKDSINSA-N Glu-Ser-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RFTVTKBHDXCEEX-WDSKDSINSA-N 0.000 description 6
- WNGHUXFWEWTKAO-YUMQZZPRSA-N Gly-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN WNGHUXFWEWTKAO-YUMQZZPRSA-N 0.000 description 6
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 6
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 6
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 6
- DTICLBJHRYSJLH-GUBZILKMSA-N Met-Ala-Val Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O DTICLBJHRYSJLH-GUBZILKMSA-N 0.000 description 6
- CAODKDAPYGUMLK-FXQIFTODSA-N Met-Asn-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O CAODKDAPYGUMLK-FXQIFTODSA-N 0.000 description 6
- 108010008355 arginyl-glutamine Proteins 0.000 description 6
- 238000010367 cloning Methods 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- FEZJJKXNPSEYEV-CIUDSAMLSA-N Arg-Gln-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O FEZJJKXNPSEYEV-CIUDSAMLSA-N 0.000 description 5
- COXMUHNBYCVVRG-DCAQKATOSA-N Arg-Leu-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O COXMUHNBYCVVRG-DCAQKATOSA-N 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- TVYMKYUSZSVOAG-ZLUOBGJFSA-N Cys-Ala-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O TVYMKYUSZSVOAG-ZLUOBGJFSA-N 0.000 description 5
- XPJBQTCXPJNIFE-ZETCQYMHSA-N Gly-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)CN XPJBQTCXPJNIFE-ZETCQYMHSA-N 0.000 description 5
- MIIVFRCYJABHTQ-ONGXEEELSA-N Gly-Leu-Val Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O MIIVFRCYJABHTQ-ONGXEEELSA-N 0.000 description 5
- JHNJNTMTZHEDLJ-NAKRPEOUSA-N Ile-Ser-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JHNJNTMTZHEDLJ-NAKRPEOUSA-N 0.000 description 5
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 5
- QYSFWUIXDFJUDW-DCAQKATOSA-N Ser-Leu-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYSFWUIXDFJUDW-DCAQKATOSA-N 0.000 description 5
- 108010061238 threonyl-glycine Proteins 0.000 description 5
- ZDILXFDENZVOTL-BPNCWPANSA-N Ala-Val-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZDILXFDENZVOTL-BPNCWPANSA-N 0.000 description 4
- 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 4
- OYTPNWYZORARHL-XHNCKOQMSA-N Gln-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N OYTPNWYZORARHL-XHNCKOQMSA-N 0.000 description 4
- CQMFNTVQVLQRLT-JHEQGTHGSA-N Gly-Thr-Gln Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O CQMFNTVQVLQRLT-JHEQGTHGSA-N 0.000 description 4
- KIZIOFNVSOSKJI-CIUDSAMLSA-N Leu-Ser-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N KIZIOFNVSOSKJI-CIUDSAMLSA-N 0.000 description 4
- UIDJDMVRDUANDL-BVSLBCMMSA-N Trp-Tyr-Arg Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UIDJDMVRDUANDL-BVSLBCMMSA-N 0.000 description 4
- BURPTJBFWIOHEY-UWJYBYFXSA-N Tyr-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 BURPTJBFWIOHEY-UWJYBYFXSA-N 0.000 description 4
- 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 4
- HTONZBWRYUKUKC-RCWTZXSCSA-N Val-Thr-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O HTONZBWRYUKUKC-RCWTZXSCSA-N 0.000 description 4
- JXWGBRRVTRAZQA-ULQDDVLXSA-N Val-Tyr-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N JXWGBRRVTRAZQA-ULQDDVLXSA-N 0.000 description 4
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 4
- 108010047495 alanylglycine Proteins 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000013604 expression vector Substances 0.000 description 4
- 108010005942 methionylglycine Proteins 0.000 description 4
- 108010029020 prolylglycine Proteins 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- 238000010200 validation analysis Methods 0.000 description 4
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 3
- NKBQZKVMKJJDLX-SRVKXCTJSA-N Arg-Glu-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NKBQZKVMKJJDLX-SRVKXCTJSA-N 0.000 description 3
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 3
- 102100031673 Corneodesmosin Human genes 0.000 description 3
- 101710139375 Corneodesmosin Proteins 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 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 3
- LGIMRDKGABDMBN-DCAQKATOSA-N Ser-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N LGIMRDKGABDMBN-DCAQKATOSA-N 0.000 description 3
- SMLCYZYQFRTLCO-UWJYBYFXSA-N Tyr-Cys-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O SMLCYZYQFRTLCO-UWJYBYFXSA-N 0.000 description 3
- NKUGCYDFQKFVOJ-JYJNAYRXSA-N Tyr-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NKUGCYDFQKFVOJ-JYJNAYRXSA-N 0.000 description 3
- DIOSYUIWOQCXNR-ONGXEEELSA-N Val-Lys-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O DIOSYUIWOQCXNR-ONGXEEELSA-N 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000002194 synthesizing effect Effects 0.000 description 3
- 108010038745 tryptophylglycine Proteins 0.000 description 3
- 108010073969 valyllysine Proteins 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- JAMAWBXXKFGFGX-KZVJFYERSA-N Ala-Arg-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JAMAWBXXKFGFGX-KZVJFYERSA-N 0.000 description 2
- QQACQIHVWCVBBR-GVARAGBVSA-N Ala-Ile-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O QQACQIHVWCVBBR-GVARAGBVSA-N 0.000 description 2
- RGQCNKIDEQJEBT-CQDKDKBSSA-N Ala-Leu-Tyr Chemical compound 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 RGQCNKIDEQJEBT-CQDKDKBSSA-N 0.000 description 2
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 description 2
- AOHKLEBWKMKITA-IHRRRGAJSA-N Arg-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AOHKLEBWKMKITA-IHRRRGAJSA-N 0.000 description 2
- 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 2
- HZPSDHRYYIORKR-WHFBIAKZSA-N Asn-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC(N)=O HZPSDHRYYIORKR-WHFBIAKZSA-N 0.000 description 2
- 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 2
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 description 2
- IHZFGJLKDYINPV-XIRDDKMYSA-N Asp-Trp-His Chemical compound C([C@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC(O)=O)N)C(O)=O)C1=CN=CN1 IHZFGJLKDYINPV-XIRDDKMYSA-N 0.000 description 2
- 208000025721 COVID-19 Diseases 0.000 description 2
- 208000028399 Critical Illness Diseases 0.000 description 2
- LZRMPXRYLLTAJX-GUBZILKMSA-N Gln-Arg-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O LZRMPXRYLLTAJX-GUBZILKMSA-N 0.000 description 2
- FQCILXROGNOZON-YUMQZZPRSA-N Gln-Pro-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O FQCILXROGNOZON-YUMQZZPRSA-N 0.000 description 2
- VYOILACOFPPNQH-UMNHJUIQSA-N Gln-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N VYOILACOFPPNQH-UMNHJUIQSA-N 0.000 description 2
- HNAUFGBKJLTWQE-IFFSRLJSSA-N Gln-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(=O)N)N)O HNAUFGBKJLTWQE-IFFSRLJSSA-N 0.000 description 2
- RQZGFWKQLPJOEQ-YUMQZZPRSA-N Gly-Arg-Gln Chemical compound C(C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)CN)CN=C(N)N RQZGFWKQLPJOEQ-YUMQZZPRSA-N 0.000 description 2
- ZOTGXWMKUFSKEU-QXEWZRGKSA-N Gly-Ile-Met Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCSC)C(O)=O ZOTGXWMKUFSKEU-QXEWZRGKSA-N 0.000 description 2
- 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 2
- 101710154606 Hemagglutinin Proteins 0.000 description 2
- XLXPYSDGMXTTNQ-DKIMLUQUSA-N Ile-Phe-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CC(C)C)C(O)=O XLXPYSDGMXTTNQ-DKIMLUQUSA-N 0.000 description 2
- XLXPYSDGMXTTNQ-UHFFFAOYSA-N Ile-Phe-Leu Natural products CCC(C)C(N)C(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=CC=C1 XLXPYSDGMXTTNQ-UHFFFAOYSA-N 0.000 description 2
- ZLFNNVATRMCAKN-ZKWXMUAHSA-N Ile-Ser-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZLFNNVATRMCAKN-ZKWXMUAHSA-N 0.000 description 2
- WGNOPSQMIQERPK-GARJFASQSA-N Leu-Asn-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N WGNOPSQMIQERPK-GARJFASQSA-N 0.000 description 2
- WGNOPSQMIQERPK-UHFFFAOYSA-N Leu-Asn-Pro Natural products CC(C)CC(N)C(=O)NC(CC(=O)N)C(=O)N1CCCC1C(=O)O WGNOPSQMIQERPK-UHFFFAOYSA-N 0.000 description 2
- AXZGZMGRBDQTEY-SRVKXCTJSA-N Leu-Gln-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O AXZGZMGRBDQTEY-SRVKXCTJSA-N 0.000 description 2
- 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 2
- GQZMPWBZQALKJO-UWVGGRQHSA-N Lys-Gly-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O GQZMPWBZQALKJO-UWVGGRQHSA-N 0.000 description 2
- CNGOEHJCLVCJHN-SRVKXCTJSA-N Lys-Pro-Glu Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O CNGOEHJCLVCJHN-SRVKXCTJSA-N 0.000 description 2
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 2
- HDNOQCZWJGGHSS-VEVYYDQMSA-N Met-Asn-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HDNOQCZWJGGHSS-VEVYYDQMSA-N 0.000 description 2
- XPVCDCMPKCERFT-GUBZILKMSA-N Met-Ser-Arg Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O XPVCDCMPKCERFT-GUBZILKMSA-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
- 108010065395 Neuropep-1 Proteins 0.000 description 2
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 2
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 2
- ZWJKVFAYPLPCQB-UNQGMJICSA-N Phe-Arg-Thr Chemical compound C[C@@H](O)[C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)Cc1ccccc1)C(O)=O ZWJKVFAYPLPCQB-UNQGMJICSA-N 0.000 description 2
- FGWUALWGCZJQDJ-URLPEUOOSA-N Phe-Thr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGWUALWGCZJQDJ-URLPEUOOSA-N 0.000 description 2
- VTFXTWDFPTWNJY-RHYQMDGZSA-N Pro-Leu-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VTFXTWDFPTWNJY-RHYQMDGZSA-N 0.000 description 2
- 101710176177 Protein A56 Proteins 0.000 description 2
- QEDMOZUJTGEIBF-FXQIFTODSA-N Ser-Arg-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O QEDMOZUJTGEIBF-FXQIFTODSA-N 0.000 description 2
- IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 description 2
- JFWDJFULOLKQFY-QWRGUYRKSA-N Ser-Gly-Phe Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JFWDJFULOLKQFY-QWRGUYRKSA-N 0.000 description 2
- RTXKJFWHEBTABY-IHPCNDPISA-N Ser-Trp-Tyr Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)NC(=O)[C@H](CO)N RTXKJFWHEBTABY-IHPCNDPISA-N 0.000 description 2
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 description 2
- DKDHTRVDOUZZTP-IFFSRLJSSA-N Thr-Gln-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DKDHTRVDOUZZTP-IFFSRLJSSA-N 0.000 description 2
- 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 2
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 2
- SVGAWGVHFIYAEE-JSGCOSHPSA-N Trp-Gly-Gln Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O)=CNC2=C1 SVGAWGVHFIYAEE-JSGCOSHPSA-N 0.000 description 2
- SLOYNOMYOAOUCX-BVSLBCMMSA-N Trp-Phe-Arg Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SLOYNOMYOAOUCX-BVSLBCMMSA-N 0.000 description 2
- BODHJXJNRVRKFA-BZSNNMDCSA-N Tyr-Cys-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BODHJXJNRVRKFA-BZSNNMDCSA-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
- ZSZFTYVFQLUWBF-QXEWZRGKSA-N Val-Asp-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCSC)C(=O)O)N ZSZFTYVFQLUWBF-QXEWZRGKSA-N 0.000 description 2
- AGKDVLSDNSTLFA-UMNHJUIQSA-N Val-Gln-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N AGKDVLSDNSTLFA-UMNHJUIQSA-N 0.000 description 2
- AJNUKMZFHXUBMK-GUBZILKMSA-N Val-Ser-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N AJNUKMZFHXUBMK-GUBZILKMSA-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
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 108010059459 arginyl-threonyl-phenylalanine Proteins 0.000 description 2
- 108010068265 aspartyltyrosine Proteins 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000002860 competitive effect Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 230000004907 flux Effects 0.000 description 2
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 2
- 108010078144 glutaminyl-glycine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 239000000185 hemagglutinin Substances 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 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 2
- 108010009298 lysylglutamic acid Proteins 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000002808 molecular sieve Substances 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 238000002823 phage display Methods 0.000 description 2
- 108010084572 phenylalanyl-valine Proteins 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000000241 respiratory effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 1
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 1
- BUDNAJYVCUHLSV-ZLUOBGJFSA-N Ala-Asp-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O BUDNAJYVCUHLSV-ZLUOBGJFSA-N 0.000 description 1
- BTYTYHBSJKQBQA-GCJQMDKQSA-N Ala-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N)O BTYTYHBSJKQBQA-GCJQMDKQSA-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
- NHLAEBFGWPXFGI-WHFBIAKZSA-N Ala-Gly-Asn Chemical compound C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)N)C(=O)O)N NHLAEBFGWPXFGI-WHFBIAKZSA-N 0.000 description 1
- AJBVYEYZVYPFCF-CIUDSAMLSA-N Ala-Lys-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O AJBVYEYZVYPFCF-CIUDSAMLSA-N 0.000 description 1
- VNFSAYFQLXPHPY-CIQUZCHMSA-N Ala-Thr-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNFSAYFQLXPHPY-CIQUZCHMSA-N 0.000 description 1
- VQBULXOHAZSTQY-GKCIPKSASA-N Ala-Trp-Phe Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O VQBULXOHAZSTQY-GKCIPKSASA-N 0.000 description 1
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 1
- 102000053723 Angiotensin-converting enzyme 2 Human genes 0.000 description 1
- PQWTZSNVWSOFFK-FXQIFTODSA-N Arg-Asp-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)CN=C(N)N PQWTZSNVWSOFFK-FXQIFTODSA-N 0.000 description 1
- LLZXKVAAEWBUPB-KKUMJFAQSA-N Arg-Gln-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LLZXKVAAEWBUPB-KKUMJFAQSA-N 0.000 description 1
- SKTGPBFTMNLIHQ-KKUMJFAQSA-N Arg-Glu-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SKTGPBFTMNLIHQ-KKUMJFAQSA-N 0.000 description 1
- DNLQVHBBMPZUGJ-BQBZGAKWSA-N Arg-Ser-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O DNLQVHBBMPZUGJ-BQBZGAKWSA-N 0.000 description 1
- YNSUUAOAFCVINY-OSUNSFLBSA-N Arg-Thr-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YNSUUAOAFCVINY-OSUNSFLBSA-N 0.000 description 1
- UGJLILSJKSBVIR-ZFWWWQNUSA-N Arg-Trp-Gly Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCN=C(N)N)N)C(=O)NCC(O)=O)=CNC2=C1 UGJLILSJKSBVIR-ZFWWWQNUSA-N 0.000 description 1
- CPTXATAOUQJQRO-GUBZILKMSA-N Arg-Val-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O CPTXATAOUQJQRO-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
- HYQYLOSCICEYTR-YUMQZZPRSA-N Asn-Gly-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O HYQYLOSCICEYTR-YUMQZZPRSA-N 0.000 description 1
- JRCASHGTXZYSPW-XIRDDKMYSA-N Asn-His-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC3=CN=CN3)NC(=O)[C@H](CC(=O)N)N JRCASHGTXZYSPW-XIRDDKMYSA-N 0.000 description 1
- QYRMBFWDSFGSFC-OLHMAJIHSA-N Asn-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QYRMBFWDSFGSFC-OLHMAJIHSA-N 0.000 description 1
- HCZQKHSRYHCPSD-IUKAMOBKSA-N Asn-Thr-Ile 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)CC)C(O)=O HCZQKHSRYHCPSD-IUKAMOBKSA-N 0.000 description 1
- XIDSGDJNUJRUHE-VEVYYDQMSA-N Asn-Thr-Met Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(O)=O XIDSGDJNUJRUHE-VEVYYDQMSA-N 0.000 description 1
- BCADFFUQHIMQAA-KKHAAJSZSA-N Asn-Thr-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BCADFFUQHIMQAA-KKHAAJSZSA-N 0.000 description 1
- JPPLRQVZMZFOSX-UWJYBYFXSA-N Asn-Tyr-Ala Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 JPPLRQVZMZFOSX-UWJYBYFXSA-N 0.000 description 1
- KSZHWTRZPOTIGY-AVGNSLFASA-N Asn-Tyr-Gln Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O KSZHWTRZPOTIGY-AVGNSLFASA-N 0.000 description 1
- BLQBMRNMBAYREH-UWJYBYFXSA-N Asp-Ala-Tyr Chemical compound N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O BLQBMRNMBAYREH-UWJYBYFXSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 1
- JJQGZGOEDSSHTE-FOHZUACHSA-N Asp-Thr-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O JJQGZGOEDSSHTE-FOHZUACHSA-N 0.000 description 1
- GCACQYDBDHRVGE-LKXGYXEUSA-N Asp-Thr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@H](O)C)NC(=O)[C@@H](N)CC(O)=O GCACQYDBDHRVGE-LKXGYXEUSA-N 0.000 description 1
- NALWOULWGHTVDA-UWVGGRQHSA-N Asp-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NALWOULWGHTVDA-UWVGGRQHSA-N 0.000 description 1
- HTSSXFASOUSJQG-IHPCNDPISA-N Asp-Tyr-Trp 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=CNC2=C1C=CC=C2)C(O)=O HTSSXFASOUSJQG-IHPCNDPISA-N 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- PKNIZMPLMSKROD-BIIVOSGPSA-N Cys-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CS)N PKNIZMPLMSKROD-BIIVOSGPSA-N 0.000 description 1
- YUZPQIQWXLRFBW-ACZMJKKPSA-N Cys-Glu-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O YUZPQIQWXLRFBW-ACZMJKKPSA-N 0.000 description 1
- NRVQLLDIJJEIIZ-VZFHVOOUSA-N Cys-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CS)N)O NRVQLLDIJJEIIZ-VZFHVOOUSA-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
- ZNTDJIMJKNNSLR-RWRJDSDZSA-N Gln-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N ZNTDJIMJKNNSLR-RWRJDSDZSA-N 0.000 description 1
- QDXMSSWCEVYOLZ-SZMVWBNQSA-N Gln-Leu-Trp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCC(=O)N)N QDXMSSWCEVYOLZ-SZMVWBNQSA-N 0.000 description 1
- LHMWTCWZARHLPV-CIUDSAMLSA-N Gln-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N LHMWTCWZARHLPV-CIUDSAMLSA-N 0.000 description 1
- MXOODARRORARSU-ACZMJKKPSA-N Glu-Ala-Ser Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)O)N MXOODARRORARSU-ACZMJKKPSA-N 0.000 description 1
- RCCDHXSRMWCOOY-GUBZILKMSA-N Glu-Arg-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O RCCDHXSRMWCOOY-GUBZILKMSA-N 0.000 description 1
- CGYDXNKRIMJMLV-GUBZILKMSA-N Glu-Arg-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O CGYDXNKRIMJMLV-GUBZILKMSA-N 0.000 description 1
- SYDJILXOZNEEDK-XIRDDKMYSA-N Glu-Arg-Trp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O SYDJILXOZNEEDK-XIRDDKMYSA-N 0.000 description 1
- GJBUAAAIZSRCDC-GVXVVHGQSA-N Glu-Leu-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O GJBUAAAIZSRCDC-GVXVVHGQSA-N 0.000 description 1
- KRRMJKMGWWXWDW-STQMWFEESA-N Gly-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)CN)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KRRMJKMGWWXWDW-STQMWFEESA-N 0.000 description 1
- DTPOVRRYXPJJAZ-FJXKBIBVSA-N Gly-Arg-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N DTPOVRRYXPJJAZ-FJXKBIBVSA-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
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- LXXANCRPFBSSKS-IUCAKERBSA-N Gly-Gln-Leu Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LXXANCRPFBSSKS-IUCAKERBSA-N 0.000 description 1
- UFPXDFOYHVEIPI-BYPYZUCNSA-N Gly-Gly-Asp Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O UFPXDFOYHVEIPI-BYPYZUCNSA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- OLPPXYMMIARYAL-QMMMGPOBSA-N Gly-Gly-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)CNC(=O)CN OLPPXYMMIARYAL-QMMMGPOBSA-N 0.000 description 1
- HAXARWKYFIIHKD-ZKWXMUAHSA-N Gly-Ile-Ser Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HAXARWKYFIIHKD-ZKWXMUAHSA-N 0.000 description 1
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 1
- LHYJCVCQPWRMKZ-WEDXCCLWSA-N Gly-Leu-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LHYJCVCQPWRMKZ-WEDXCCLWSA-N 0.000 description 1
- BXICSAQLIHFDDL-YUMQZZPRSA-N Gly-Lys-Asn Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BXICSAQLIHFDDL-YUMQZZPRSA-N 0.000 description 1
- FHQRLHFYVZAQHU-IUCAKERBSA-N Gly-Lys-Gln Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O FHQRLHFYVZAQHU-IUCAKERBSA-N 0.000 description 1
- PDUHNKAFQXQNLH-ZETCQYMHSA-N Gly-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)NCC(O)=O PDUHNKAFQXQNLH-ZETCQYMHSA-N 0.000 description 1
- OJNZVYSGVYLQIN-BQBZGAKWSA-N Gly-Met-Asp Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(O)=O OJNZVYSGVYLQIN-BQBZGAKWSA-N 0.000 description 1
- 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 1
- JPVGHHQGKPQYIL-KBPBESRZSA-N Gly-Phe-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 JPVGHHQGKPQYIL-KBPBESRZSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- 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
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- WCORRBXVISTKQL-WHFBIAKZSA-N Gly-Ser-Ser Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O WCORRBXVISTKQL-WHFBIAKZSA-N 0.000 description 1
- UMRIXLHPZZIOML-OALUTQOASA-N Gly-Trp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)CN UMRIXLHPZZIOML-OALUTQOASA-N 0.000 description 1
- IROABALAWGJQGM-OALUTQOASA-N Gly-Trp-Tyr Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)NC(=O)CN IROABALAWGJQGM-OALUTQOASA-N 0.000 description 1
- 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
- KSOBNUBCYHGUKH-UWVGGRQHSA-N Gly-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN KSOBNUBCYHGUKH-UWVGGRQHSA-N 0.000 description 1
- 101710114810 Glycoprotein Proteins 0.000 description 1
- 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 1
- AHEBIAHEZWQVHB-QTKMDUPCSA-N His-Thr-Met Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N)O AHEBIAHEZWQVHB-QTKMDUPCSA-N 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- HDOYNXLPTRQLAD-JBDRJPRFSA-N Ile-Ala-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)O)N HDOYNXLPTRQLAD-JBDRJPRFSA-N 0.000 description 1
- NCSIQAFSIPHVAN-IUKAMOBKSA-N Ile-Asn-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N NCSIQAFSIPHVAN-IUKAMOBKSA-N 0.000 description 1
- UYNXBNHVWFNVIN-HJWJTTGWSA-N Ile-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)CC)CC1=CC=CC=C1 UYNXBNHVWFNVIN-HJWJTTGWSA-N 0.000 description 1
- SAEWJTCJQVZQNZ-IUKAMOBKSA-N Ile-Thr-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N SAEWJTCJQVZQNZ-IUKAMOBKSA-N 0.000 description 1
- PRTZQMBYUZFSFA-XEGUGMAKSA-N Ile-Tyr-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)NCC(=O)O)N PRTZQMBYUZFSFA-XEGUGMAKSA-N 0.000 description 1
- GVEODXUBBFDBPW-MGHWNKPDSA-N Ile-Tyr-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C(O)=O)CC1=CC=C(O)C=C1 GVEODXUBBFDBPW-MGHWNKPDSA-N 0.000 description 1
- APQYGMBHIVXFML-OSUNSFLBSA-N Ile-Val-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N APQYGMBHIVXFML-OSUNSFLBSA-N 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- VPKIQULSKFVCSM-SRVKXCTJSA-N Leu-Gln-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VPKIQULSKFVCSM-SRVKXCTJSA-N 0.000 description 1
- JNDYEOUZBLOVOF-AVGNSLFASA-N Leu-Leu-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O JNDYEOUZBLOVOF-AVGNSLFASA-N 0.000 description 1
- BIZNDKMFQHDOIE-KKUMJFAQSA-N Leu-Phe-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(O)=O)CC1=CC=CC=C1 BIZNDKMFQHDOIE-KKUMJFAQSA-N 0.000 description 1
- WFCKERTZVCQXKH-KBPBESRZSA-N Leu-Tyr-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O WFCKERTZVCQXKH-KBPBESRZSA-N 0.000 description 1
- CANPXOLVTMKURR-WEDXCCLWSA-N Lys-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN CANPXOLVTMKURR-WEDXCCLWSA-N 0.000 description 1
- PNDCUTDWYVKBHX-IHRRRGAJSA-N Met-Asp-Tyr Chemical compound CSCC[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 PNDCUTDWYVKBHX-IHRRRGAJSA-N 0.000 description 1
- LXCSZPUQKMTXNW-BQBZGAKWSA-N Met-Ser-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O LXCSZPUQKMTXNW-BQBZGAKWSA-N 0.000 description 1
- CQRGINSEMFBACV-WPRPVWTQSA-N Met-Val-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O CQRGINSEMFBACV-WPRPVWTQSA-N 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 1
- GPLWGAYGROGDEN-BZSNNMDCSA-N Phe-Phe-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O GPLWGAYGROGDEN-BZSNNMDCSA-N 0.000 description 1
- MGDFPGCFVJFITQ-CIUDSAMLSA-N Pro-Glu-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MGDFPGCFVJFITQ-CIUDSAMLSA-N 0.000 description 1
- 241000315672 SARS coronavirus Species 0.000 description 1
- AEGUWTFAQQWVLC-BQBZGAKWSA-N Ser-Gly-Arg Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O AEGUWTFAQQWVLC-BQBZGAKWSA-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
- JEHPKECJCALLRW-CUJWVEQBSA-N Ser-His-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JEHPKECJCALLRW-CUJWVEQBSA-N 0.000 description 1
- SFTZTYBXIXLRGQ-JBDRJPRFSA-N Ser-Ile-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O SFTZTYBXIXLRGQ-JBDRJPRFSA-N 0.000 description 1
- UIPXCLNLUUAMJU-JBDRJPRFSA-N Ser-Ile-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O UIPXCLNLUUAMJU-JBDRJPRFSA-N 0.000 description 1
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 1
- SQHKXWODKJDZRC-LKXGYXEUSA-N Ser-Thr-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQHKXWODKJDZRC-LKXGYXEUSA-N 0.000 description 1
- KKKVOZNCLALMPV-XKBZYTNZSA-N Ser-Thr-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O KKKVOZNCLALMPV-XKBZYTNZSA-N 0.000 description 1
- SNXUIBACCONSOH-BWBBJGPYSA-N Ser-Thr-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CO)C(O)=O SNXUIBACCONSOH-BWBBJGPYSA-N 0.000 description 1
- BCAVNDNYOGTQMQ-AAEUAGOBSA-N Ser-Trp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)NCC(O)=O BCAVNDNYOGTQMQ-AAEUAGOBSA-N 0.000 description 1
- PIQRHJQWEPWFJG-UWJYBYFXSA-N Ser-Tyr-Ala 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(O)=O PIQRHJQWEPWFJG-UWJYBYFXSA-N 0.000 description 1
- FGBLCMLXHRPVOF-IHRRRGAJSA-N Ser-Tyr-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FGBLCMLXHRPVOF-IHRRRGAJSA-N 0.000 description 1
- 101710167605 Spike glycoprotein Proteins 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 1
- XSLXHSYIVPGEER-KZVJFYERSA-N Thr-Ala-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O XSLXHSYIVPGEER-KZVJFYERSA-N 0.000 description 1
- JVTHIXKSVYEWNI-JRQIVUDYSA-N Thr-Asn-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JVTHIXKSVYEWNI-JRQIVUDYSA-N 0.000 description 1
- LKEKWDJCJSPXNI-IRIUXVKKSA-N Thr-Glu-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 LKEKWDJCJSPXNI-IRIUXVKKSA-N 0.000 description 1
- QQWNRERCGGZOKG-WEDXCCLWSA-N Thr-Gly-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O QQWNRERCGGZOKG-WEDXCCLWSA-N 0.000 description 1
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 1
- 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
- ABWNZPOIUJMNKT-IXOXFDKPSA-N Thr-Phe-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O ABWNZPOIUJMNKT-IXOXFDKPSA-N 0.000 description 1
- MXNAOGFNFNKUPD-JHYOHUSXSA-N Thr-Phe-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MXNAOGFNFNKUPD-JHYOHUSXSA-N 0.000 description 1
- WPSKTVVMQCXPRO-BWBBJGPYSA-N Thr-Ser-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O WPSKTVVMQCXPRO-BWBBJGPYSA-N 0.000 description 1
- HUPLKEHTTQBXSC-YJRXYDGGSA-N Thr-Ser-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HUPLKEHTTQBXSC-YJRXYDGGSA-N 0.000 description 1
- PWONLXBUSVIZPH-RHYQMDGZSA-N Thr-Val-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O PWONLXBUSVIZPH-RHYQMDGZSA-N 0.000 description 1
- KWAIRYFEWMLWPQ-UHFFFAOYSA-N Trp Gly Ser Tyr Chemical compound C=1NC2=CC=CC=C2C=1CC(N)C(=O)NCC(=O)NC(CO)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 KWAIRYFEWMLWPQ-UHFFFAOYSA-N 0.000 description 1
- MHNHRNHJMXAVHZ-AAEUAGOBSA-N Trp-Asn-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N MHNHRNHJMXAVHZ-AAEUAGOBSA-N 0.000 description 1
- WLBZWXXGSOLJBA-HOCLYGCPSA-N Trp-Gly-Lys Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(=O)N[C@@H](CCCCN)C(O)=O)=CNC2=C1 WLBZWXXGSOLJBA-HOCLYGCPSA-N 0.000 description 1
- WNZRNOGHEONFMS-PXDAIIFMSA-N Trp-Ile-Tyr Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O WNZRNOGHEONFMS-PXDAIIFMSA-N 0.000 description 1
- CDRYEAWHKJSGAF-BPNCWPANSA-N Tyr-Ala-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O CDRYEAWHKJSGAF-BPNCWPANSA-N 0.000 description 1
- XBWKCYFGRXKWGO-SRVKXCTJSA-N Tyr-Cys-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O XBWKCYFGRXKWGO-SRVKXCTJSA-N 0.000 description 1
- QOEZFICGUZTRFX-IHRRRGAJSA-N Tyr-Cys-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O QOEZFICGUZTRFX-IHRRRGAJSA-N 0.000 description 1
- NGALWFGCOMHUSN-AVGNSLFASA-N Tyr-Gln-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NGALWFGCOMHUSN-AVGNSLFASA-N 0.000 description 1
- OLWFDNLLBWQWCP-STQMWFEESA-N Tyr-Gly-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O OLWFDNLLBWQWCP-STQMWFEESA-N 0.000 description 1
- SMUWZUSWMWVOSL-JYJNAYRXSA-N Tyr-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N SMUWZUSWMWVOSL-JYJNAYRXSA-N 0.000 description 1
- IZFVRRYRMQFVGX-NRPADANISA-N Val-Ala-Gln Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N IZFVRRYRMQFVGX-NRPADANISA-N 0.000 description 1
- ASQFIHTXXMFENG-XPUUQOCRSA-N Val-Ala-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O ASQFIHTXXMFENG-XPUUQOCRSA-N 0.000 description 1
- JVYIGCARISMLMV-HOCLYGCPSA-N Val-Gly-Trp Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N JVYIGCARISMLMV-HOCLYGCPSA-N 0.000 description 1
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 1
- JQTYTBPCSOAZHI-FXQIFTODSA-N Val-Ser-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N JQTYTBPCSOAZHI-FXQIFTODSA-N 0.000 description 1
- PQSNETRGCRUOGP-KKHAAJSZSA-N Val-Thr-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O PQSNETRGCRUOGP-KKHAAJSZSA-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
- 230000001154 acute effect Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 238000009175 antibody therapy Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000005560 droplet transmission Effects 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 210000003000 inclusion body Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000012482 interaction analysis Methods 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000004001 molecular interaction Effects 0.000 description 1
- 108010072637 phenylalanyl-arginyl-phenylalanine Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000007671 third-generation sequencing Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000007501 viral attachment Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 238000001262 western blot Methods 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/567—Framework region [FR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/10—Detection of antigens from microorganism in sample from host
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Hematology (AREA)
- General Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Oncology (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Communicable Diseases (AREA)
- General Physics & Mathematics (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pulmonology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The embodiment of the invention discloses a nano antibody for targeting a novel coronavirus and a preparation method and application thereof, wherein the nano antibody comprises at least one of a nano antibody NbN2, a nano antibody NbN3, a nano antibody NbN4, a nano antibody NbN5, a nano antibody NbN6 and a nano antibody NbN11, and the provided nano antibody has higher affinity to RBD structural domains of a novel coronavirus wild strain, a novel coronavirus variant Alpha (Alpha), a novel coronavirus Beta (Beta), a novel coronavirus Delta (Delta), a novel coronavirus omacron and subtype BA.2 thereof, can rapidly and specifically neutralize different novel coronaviruses, and effectively inhibit the activity of the novel coronavirus; meanwhile, the provided nano antibody has a longer CDR3 sequence, is beneficial to flexibly regulating the conformation of the nano antibody, improves the binding effect with viruses, and is beneficial to wide application.
Description
The application is a divisional application of a nanometer antibody targeting a new coronavirus, a preparation method and application thereof, the application date of the original application is 2022, 5 and 23, the application number is 202210330386.7, and the invention creates the nanometer antibody targeting the new coronavirus, and the preparation method and application thereof.
Technical Field
The invention relates to the technical field of biology, in particular to a nano antibody targeting a novel coronavirus, a preparation method and application thereof.
Background
The novel coronavirus pneumonia (Corona Virus Disease 2019, covd-19) can be transmitted through the approaches of droplet transmission, contact with respiratory secretions, aerosol transmission and the like, and the consequences are serious. The acute respiratory syndrome coronavirus 2 (Severe acute respiratory syndrome coronavirus, SARS-CoV-2) causing the COVID-19 expresses mainly 4 structural proteins, spike (S), envelope (envelope), membrane (membrane) and nucleocapsid (nucleocapsid), respectively. Wherein the S protein comprises an N-terminal S1 subunit and a C-terminal S2 subunit, the S1 subunit comprising a receptor binding domain of about 200 amino acids (receptor binding domain, RBD) being responsible for binding of the virus to the host receptor and the S2 subunit being responsible for fusion of the virus to the host cell membrane. The S protein plays a vital role in virus attachment, infection and transmission, and is an important target for the research and development of anti-SARS-CoV-2 drugs by being combined with host cell angiotensin converting enzyme2 (angiotensin converting enzyme, ACE 2) to infect cells.
Neutralizing antibodies are an important therapy against covd-19. It has been found that clinical outcome can be significantly improved by delivering serum from a convalescent patient to a critically ill covd-19 patient, indicating that neutralizing antibody therapy also has better relief from new coronary critically ill patients. The mechanism of action is that the neutralizing antibody binds with spike glycoprotein on the surface of the novel coronavirus to further influence the binding of SARS-CoV-2 with the surface receptor of the host cell, thereby preventing the virus from infecting the target cell. There are a number of neutralizing antibodies currently marketed for the treatment of covd-19, and these monoclonal antibody targets are primarily focused on the RBD domain of the S protein. However, in the face of constantly mutated SARS-CoV-2, it is difficult for existing neutralizing antibodies to target new novel coronavariants. A recent study showed that omacron subtype BA.2 could evade almost all neutralizing antibody treatments. Thus, there is an urgent need to develop new neutralizing antibodies that can simultaneously target different novel crown variants, particularly omacron and its subtype ba.2.
Disclosure of Invention
The application aims to provide a nano antibody targeting a novel coronavirus, a preparation method and application thereof, and aims to solve the problem that a neutralizing antibody of a novel coronavirus variant Omicron and a subtype BA.2 thereof is lacking in the prior art.
In order to achieve the purposes of the application, the technical scheme adopted by the application is as follows:
in a first aspect, the application discloses a nanobody for targeting a novel coronavirus, wherein the nanobody comprises at least one of nanobody NbN2, nanobody NbN3, nanobody NbN4, nanobody NbN5, nanobody NbN6 and nanobody NbN11, wherein the amino acid sequence of nanobody NbN2 is shown as seq. ID No.1, the amino acid sequence of nanobody NbN3 is shown as seq. ID No.2, the amino acid sequence of nanobody NbN4 is shown as seq. ID No.3, the amino acid sequence of nanobody NbN5 is shown as seq. ID No.4, and the amino acid sequence of nanobody NbN6 is shown as seq. ID No.5, and the amino acid sequence of nanobody NbN11 is shown as seq. ID No. 6.
In a second aspect, the present application discloses a method for preparing a nanobody targeting a novel coronavirus, comprising the steps of:
providing target proteins, coating the target proteins on an immune tube, and carrying out enrichment screening to obtain a phage library;
performing second generation sequencing on the eluent of the phage library, and synthesizing a gene sequence of the nano antibody with the original count ratio of 10 according to a sequencing result;
cloning the gene sequence of the nano antibody into an expression vector to obtain a recombinant plasmid, transferring the recombinant plasmid into a host cell to induce expression and purifying to obtain the nano antibody of the targeted novel coronavirus.
In a third aspect, the application discloses the use of nanobodies targeting novel coronaviruses in the preparation of a medicament for the prevention and/or treatment of novel coronavirus pneumonia.
In a fourth aspect, the application discloses application of a nanobody targeting a novel coronavirus in preparing a detection reagent or a detection kit for detecting the novel coronavirus.
The nanometer antibody of the novel coronavirus Omicron and subtype BA.2 provided in the first aspect of the application comprises at least one of a nanometer antibody NbN2, a nanometer antibody NbN3, a nanometer antibody NbN4, a nanometer antibody NbN5, a nanometer antibody NbN6 and a nanometer antibody NbN11, and the provided nanometer antibody has higher affinity to RBD domains of a wild strain of the novel coronavirus, a variant Alpha (Alpha) of the novel coronavirus, a Beta (Beta) of the novel coronavirus, delta (Delta) of the novel coronavirus, the Omicron of the novel coronavirus and subtype BA.2 thereof, can neutralize different novel coronaviruses rapidly and specifically, and effectively inhibit the activity of the novel coronavirus; meanwhile, the provided nano antibody has a longer CDR3 sequence, is beneficial to flexibly regulating the conformation of the nano antibody, improves the binding effect with viruses, has lower preparation cost and is beneficial to wide application.
According to the preparation method of the nanometer antibody targeting the novel coronavirus Omicron and the subtype BA.2 thereof, which is provided by the second aspect of the application, a nanometer antibody library is constructed by taking RBD proteins of the novel coronavirus Omicron and the subtype BA.2 thereof as targets based on phage display screening technology, and the nanometer antibody targeting the novel coronavirus Omicron and the subtype BA.2 thereof is obtained by combining second-generation sequencing, sequence synthesis, cloning, expression and purification; the preparation method utilizes the biological characteristics of phage fast replication in host bacteria, can rapidly screen the nano antibody combined with the specific variant Omikovia and subtype BA.2RBD protein thereof with high flux, is rapid and simple, is favorable for large-scale screening, and improves the screening efficiency.
The application of the nanobody of the targeting novel coronavirus Omicron and subtype BA.2 thereof in preparing the medicine for preventing and/or treating the novel coronavirus pneumonia is provided in the third aspect of the application, and the nanobody of the targeting novel coronavirus Omicron and subtype BA.2 thereof obtained by screening comprises at least one of nanobody NbN2, nanobody NbN3, nanobody NbN4, nanobody NbN5, nanobody NbN6 and nanobody NbN11, has better neutralization activity capability and inhibition activity capability on viruses of wild strains and variant strains of the novel coronavirus, and is suitable for preparing related medicines for preventing and/or treating the novel coronavirus pneumonia.
The application of the nanometer antibody for targeting the novel coronavirus Omicron and the subtype BA.2 thereof in preparing the detection reagent or the detection kit for detecting the novel coronavirus is provided, the nanometer antibody for targeting the novel coronavirus Omicron and the subtype BA.2 thereof comprises at least one of nanometer antibody NbN2, nanometer antibody NbN3, nanometer antibody NbN4, nanometer antibody NbN5, nanometer antibody NbN6 and nanometer antibody NbN11, has higher binding capacity for the wild strain of the novel coronavirus and the viruses of various variants, and is suitable for preparing the detection reagent or the detection kit for detecting the novel coronavirus.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Wherein:
FIG. 1 is a schematic diagram of Omicron RBD protein nanobody screening.
FIG. 2 is a schematic representation of the sequence of third generation sequencing top 10 of phage eluate from Omicron RBD screening.
FIG. 3 is a schematic representation of Omicron RBD nanobody expression purification.
FIG. 4 is a schematic diagram of nanobody immunoblotting (Western Blot, WB) detection.
FIG. 5 is a schematic diagram of ELISA validation of Omacron RBD nanobodies.
FIG. 6 is a schematic diagram of ELISA validation of SARS-CoV-2 Wild-Type (Wild Type, WT) RBD nanobody.
FIG. 7 is a schematic of ELISA validation of Delta RBD nanobodies.
FIG. 8 is a schematic diagram of ELISA validation of nanobody and negative control protein bovine serum albumin (Bovine Serum Albumin, BSA).
FIG. 9 is a schematic of affinity constants of Biacore detection nanobody N2 and WT RBD proteins.
FIG. 10 is a schematic diagram showing affinity constants of the Biacore detection nanobody N2 and Alpha RBD proteins.
FIG. 11 is a schematic of the affinity constants of Biacore detection nanobody N2 and Beta RBD protein.
FIG. 12 is a schematic of the affinity constants of Biacore detection nanobody N2 and Delta RBD proteins.
FIG. 13 is a schematic diagram showing affinity constants of Biacore detection nanobody N2 and Omicron RBD protein.
FIG. 14 is a schematic representation of the affinity constants of Biacore detection nanobody N2 to Omicron BA.2RBD protein.
FIG. 15 is a schematic of the affinity constants of Biacore detection nanobody N3 and WT RBD proteins.
FIG. 16 is a schematic of the affinity constants of Biacore detection nanobody N3 and Alpha RBD protein.
FIG. 17 is a schematic of the affinity constants of Biacore detection nanobody N3 and Beta RBD protein.
FIG. 18 is a schematic of the affinity constants of Biacore detection nanobody N3 and Delta RBD protein.
FIG. 19 is a schematic of affinity constants of Biacore detection nanobody N3 and Omicron RBD protein.
FIG. 20 is a schematic representation of the affinity constants of Biacore detection nanobody N3 to Omicron BA.2RBD protein.
FIG. 21 is a schematic of the affinity constants of Biacore detection nanobody N4 to WT RBD protein.
FIG. 22 is a schematic of affinity constants of Biacore detection nanobody N4 and Alpha RBD protein.
FIG. 23 is a schematic representation of the affinity constants of Biacore detection nanobody N4 and Beta RBD protein.
FIG. 24 is a schematic of the affinity constants of Biacore detection nanobody N4 and Delta RBD protein.
FIG. 25 is a schematic representation of the affinity constants of Biacore detection nanobody N4 to Omicron RBD protein.
FIG. 26 is a schematic of the affinity constants of Biacore detection nanobody N4 with Omicron BA.2RBD protein.
FIG. 27 is a schematic of the affinity constants of Biacore detection nanobody N5 to WT RBD protein.
FIG. 28 is a schematic of the affinity constants of Biacore detection nanobody N5 and Alpha RBD protein.
FIG. 29 is a schematic of the affinity constants of Biacore detection nanobody N5 and Beta RBD protein.
FIG. 30 is a schematic diagram showing affinity of Biacore detection nanobody N5 with Delta RBD protein.
FIG. 31 is a schematic diagram showing affinity constants of Biacore detection nanobody N5 and Omicron RBD protein.
FIG. 32 is a schematic of affinity constants of Biacore detection nanobody N5 with Omicron BA.2RBD protein.
FIG. 33 is a schematic of the affinity constants of Biacore detection nanobody N6 to WT RBD protein.
FIG. 34 is a schematic of the affinity constants of Biacore detection nanobody N6 and Alpha RBD protein.
FIG. 35 is a schematic of the affinity constants of Biacore detection nanobody N6 and Beta RBD protein.
FIG. 36 is a schematic of the affinity constants of Biacore detection nanobody N6 and Delta RBD protein.
FIG. 37 is a schematic representation of the affinity constants of Biacore detection nanobody N6 to Omicron RBD protein.
FIG. 38 is a schematic of the affinity constants of Biacore detection nanobody N6 to Omicron BA.2RBD protein.
FIG. 39 is a schematic of the affinity constants of Biacore detection nanobody N11 to WT RBD proteins.
FIG. 40 is a schematic of the affinity constants of Biacore detection nanobody N11 and Alpha RBD protein.
FIG. 41 is a schematic of the affinity constants of Biacore detection nanobody N11 and Beta RBD protein.
FIG. 42 is a schematic of the affinity constants of Biacore detection nanobody N11 and Delta RBD protein.
FIG. 43 is a schematic of affinity constants of Biacore detection nanobody N11 with Omicron RBD protein.
FIG. 44 is a schematic of the affinity constants of Biacore detection nanobody N11 to Omicron BA.2RBD protein.
Figure 45 is a schematic of a nanobody and ACE2 competition ELISA.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The nanobody of the targeting novel coronavirus provided in the first aspect of the application includes at least one of nanobody NbN2, nanobody NbN3, nanobody NbN4, nanobody NbN5, nanobody NbN6 and nanobody NbN11, wherein the amino acid sequence of nanobody NbN2 is shown as seq.id No.1, the amino acid sequence of nanobody NbN3 is shown as seq.id No.2, the amino acid sequence of nanobody NbN4 is shown as seq.id No.3, the amino acid sequence of nanobody NbN5 is shown as seq.id No.4, the amino acid sequence of nanobody NbN6 is shown as seq.id No.5, and the amino acid sequence of nanobody NbN11 is shown as seq.id No. 6.
The nanometer antibody of the novel coronavirus Omicron and subtype BA.2 thereof provided in the first aspect of the application comprises at least one of nanometer antibody NbN2, nanometer antibody NbN3, nanometer antibody NbN4, nanometer antibody NbN5, nanometer antibody NbN6 and nanometer antibody NbN11, and the provided nanometer antibody has higher affinity to RBD domains of novel coronavirus wild strains, novel coronavirus variant Alpha (Alpha), novel coronavirus Beta (Beta), novel coronavirus Delta (Delta), novel coronavirus Omicron and subtype BA.2 thereof, can rapidly and specifically neutralize different novel coronaviruses, and effectively inhibit the activity of the novel coronavirus; meanwhile, the provided nano antibody has a longer CDR3 sequence, is beneficial to flexibly regulating the conformation of the nano antibody, improves the binding effect with viruses, has lower preparation cost and is beneficial to wide application.
The amino acid sequence of the nanobody NbN2 is shown as seq. ID No.1, and the seq. ID No.1 is as follows: MAVQLVESGGGLVQAGGSLRLSCAATGLTFTGYVMAWFRQAPGKNREFVAAV SRSGVVTRYADSVKGRFTVSRDNAKNTVYLQMNTLNPEDTALYYCAADWHP LTGIMTTDSTEYDYWGQGTQVTVSS.
The base sequence of nanobody NbN2 is shown as seq. ID No.44, and seq. ID No.44 is as follows: ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGATTGGTGCAGGCTGGGGG CTCTCTGAGACTCTCCTGTGCAGCCACTGGACTCACCTTCACTGGTTATGTA ATGGCCTGGTTCCGCCAGGCTCCAGGGAAGAACCGTGAGTTTGTAGCGGCC GTTAGCCGGAGTGGTGTTGTTACACGCTATGCAGACTCCGTGAAGGGCCGA TTCACCGTCTCCAGAGACAACGCCAAGAACACGGTGTATCTGCAAATGAAC ACCCTGAATCCTGAGGACACGGCCCTTTATTACTGTGCAGCAGATTGGCATC CACTAACTGGAATAATGACGACTGACTCTACAGAGTATGACTACTGGGGCC AGGGGACCCAGGTCACCGTCTCCTCA.
The amino acid sequence of the nanobody NbN3 is shown as seq. ID No.2, and the seq. ID No.2 is as follows: MAVQLVESGGGLVQAGGSLRLSCAASGRTFSRYAMGWFRQAPGKERQFVAGI SGSGIVTNYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAIYYCAPLILARTV DMNHWGKGTQVTVSS.
The base sequence of nanobody NbN3 is shown as seq. ID No.45, and seq. ID No.45 is as follows: ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGATTGGTGCAGGCTGGGGG CTCTCTGAGACTCTCCTGTGCAGCCTCTGGACGCACCTTCAGTAGGTATGCC ATGGGCTGGTTCCGCCAGGCTCCAGGGAAGGAGCGCCAGTTTGTAGCAGGT ATTAGCGGGAGTGGTATAGTCACAAACTATGCAGACTCCGTGAAGGGCCGA TTCACCATCTCCAGAGACAACGCCAAGAACACGGTGTACCTGCAAATGAAC AGCCTGAAACCTGAGGACACGGCCATTTATTACTGTGCACCGTTGATCCTAG CACGAACTGTGGACATGAACCACTGGGGCAAAGGGACCCAGGTCACCGTC TCCTCA.
The amino acid sequence of the nanobody NbN4 is shown as seq. ID No.3, and the sequence of the seq. ID No.3 is as follows: MAVQLVESGGGVVQPGGSLRLSCTASGFGFSLYGMSWYRQAPGKERELVASIA SGGDSTNYQDSVKGRFSMSRDNAKSTVYLQMNSLKPEDTAVYYCVAERVSSF LKIRTAYWGQGTQVTVSS.
The base sequence of nanobody NbN4 is shown as seq. ID No. 46. Seq. ID No.46 is as follows: ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGAGTGGTGCAGCCTGGGGG GTCTCTGAGACTCTCCTGTACAGCCTCTGGATTCGGCTTCAGTCTTTATGGC ATGAGCTGGTACCGTCAGGCTCCAGGGAAGGAGCGCGAGTTGGTCGCATCT ATTGCCAGTGGTGGTGATAGTACAAACTATCAAGACTCCGTGAAGGGCCGA TTCTCCATGTCCAGAGACAATGCCAAGAGCACGGTGTATCTGCAAATGAAC AGCCTGAAACCTGAGGACACGGCCGTGTATTACTGCGTGGCAGAAAGAGT CTCGTCGTTCTTAAAAATTCGTACCGCCTACTGGGGCCAGGGGACCCAGGT CACCGTCTCCTCA.
The amino acid sequence of the nanobody NbN5 is shown as seq. ID No.4, and the sequence of the seq. ID No.4 is as follows: MAVQLVESGGGLVQAGGSLRVSCEASGFLFNGDAYVLGWYRQVPGRQRELVA TITSSGDTSYADSVKGRFTISRDNAKNTIFLQMSGLKPEDTAVYYCYAERWNGL LQRWGQGTQVTVSS.
The base sequence of nanobody NbN5 is shown as seq. ID No.47, and seq. ID No.47 is as follows: ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAGGCTGGGGG GTCTCTGAGAGTGTCCTGTGAAGCCTCTGGATTCCTTTTCAATGGCGATGCC TATGTCTTGGGCTGGTACCGCCAGGTTCCAGGTAGGCAGCGTGAATTGGTC GCAACTATTACGAGTAGCGGTGACACAAGCTATGCGGACTCCGTGAAGGGC CGATTCACCATCTCCAGAGACAACGCCAAGAATACAATTTTTCTGCAAATG AGCGGCCTGAAACCCGAGGACACGGCCGTCTATTACTGCTATGCAGAGAGG TGGAATGGGCTCCTTCAACGCTGGGGCCAGGGGACCCAGGTCACCGTCTCC TCA.
The amino acid sequence of the nanobody NbN6 is shown as seq. ID No.5, and the sequence of the seq. ID No.5 is as follows: MAVQLVESGGGLVQPGGSLRLSCAASGIIFRFRTISWYRQAPGKQRELVASISG GSSTSYADTVKGRFTISRDNAKNTVYLQMNSLKPEDTGVYYCAASHTMVGWI YGMDYWGKGTQVTVSS.
The base sequence of nanobody NbN6 is shown as seq. ID No.48, and seq. ID No.48 is as follows: ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAACCTGGGGG GTCTCTGAGACTCTCCTGTGCAGCCTCTGGAATCATCTTCAGATTCAGAACC ATATCCTGGTACCGCCAGGCTCCAGGGAAGCAGCGCGAGTTGGTCGCAAGT ATTAGTGGCGGCAGTAGCACGTCCTATGCAGACACTGTGAAGGGCCGATTC ACCATCTCCAGAGACAACGCCAAGAACACGGTGTATCTGCAAATGAACAG CCTGAAACCTGAGGACACAGGCGTTTATTACTGTGCAGCGTCACATACTATG GTTGGGTGGATCTACGGCATGGACTACTGGGGCAAAGGGACCCAGGTCACC GTCTCCTCA.
The amino acid sequence of the nanobody NbN11 is shown as seq. ID No.6, and the sequence of the seq. ID No.6 is as follows: MAVQLVESGGGLVQAGGSLRLSCAASGSFFSINTMGWYRQAPGKQRELVAQIT NAGNTNYADSVKGRFTISRDNAKNTIYLQMNSLKPEDTAVYYCNAGQLWGSY RSWGQGTQVTVSS.
The base sequence of nanobody NbN11 is shown as seq. ID No.49, and seq. ID No.49 is as follows: ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAGGCTGGGGG GTCTCTGAGACTCTCCTGTGCAGCCTCTGGAAGCTTCTTCAGTATCAATACC ATGGGCTGGTACCGCCAGGCTCCAGGGAAGCAGCGCGAGTTGGTCGCACA AATTACTAATGCAGGTAACACAAACTATGCAGACTCCGTGAAGGGCCGGTT CACCATCTCCAGAGACAACGCCAAGAACACGATATATCTGCAAATGAACAG CCTGAAACCTGAGGACACGGCCGTCTATTACTGCAATGCAGGACAGTTGTG GGGTAGTTACCGCTCCTGGGGCCAGGGGACCCAGGTCACCGTCTCCTCA.
In some embodiments, nanobodies include 4 framework regions FR1, FR2, FR3, FR4, and 3 complementarity determining regions CDR1, CDR2, CDR3; the difference in amino acid sequences of the respective regions affects the binding ability of the nanobody to viruses.
In the nano antibody NbN2, the amino acid sequence of FR1 is shown as SEQ ID NO.7, and specifically comprises the following steps: AVQLVESGGGLVQAGGSLRLSCAATGLTFT; the amino acid sequence of FR2 is shown as SEQ ID NO.8, and specifically comprises the following steps: WFRQAPGKNREFVA; the amino acid sequence of FR3 is shown as SEQ ID NO.9, and specifically comprises the following steps: RFTVSRDNAKNTVYLQMNTLNPEDTALYYCAA; the amino acid sequence of FR4 is shown as SEQ ID NO.10, and specifically comprises the following steps: WGQGTQVTVSS; the amino acid sequence of CDR1 is shown in SEQ ID NO.11, specifically: GYVMA; the amino acid sequence of CDR2 is shown in SEQ ID NO.12, specifically: AVSRSGVVTRYADSVKG; the amino acid sequence of CDR3 is shown in SEQ ID NO.13, specifically: DWHPLTGIMTTDSTEYDY.
In the nano antibody NbN3, the amino acid sequence of FR1 is shown as SEQ ID NO.14, and specifically comprises the following steps: AVQLVESGGGLVQAGGSLRLSCAASGRTFS; the amino acid sequence of FR2 is shown as SEQ ID NO.15, and specifically comprises the following steps: WFRQAPGKERQFVA; the amino acid sequence of FR3 is shown as SEQ ID NO.16, and specifically comprises the following steps: RFT ISRDNAKNTVYLQMNSLKPEDTA I YYCAP; the amino acid sequence of FR4 is shown as SEQ ID NO.17, specifically WGKGTQVTVSS; the amino acid sequence of CDR1 is shown as SEQ ID NO.18, specifically: RYAMG; the amino acid sequence of CDR2 is shown in SEQ ID NO.19, specifically: GI SGSGI VTNYADSVKG; the amino acid sequence of CDR3 is shown in SEQ ID NO.20, specifically: LILARTVDMNH.
In the nano antibody NbN4, the amino acid sequence of FR1 is shown as SEQ ID NO.21, and specifically comprises the following steps: AVQLVESGGGVVQPGGSLRLSCTASGFGFS; the amino acid sequence of FR2 is shown as SEQ ID NO.22, and specifically comprises the following steps: WYRQAPGKERELVA; the amino acid sequence of FR3 is shown as SEQ ID NO.23, and specifically comprises the following steps: RFSMSRDNAKSTVYLQMNSLKPEDTAVYYCVA; the amino acid sequence of FR4 is shown as SEQ ID NO.10, and specifically comprises the following steps: WGQGTQVTVSS; the amino acid sequence of CDR1 is shown in SEQ ID NO.24, specifically: LYGMS; the amino acid sequence of CDR2 is shown in SEQ ID NO.25, specifically: SI ASGGDSTNYQDSVKG; the amino acid sequence of CDR3 is shown in SEQ ID NO.26, specifically: ERVSSFL K I RTAY.
In the nano antibody NbN5, the amino acid sequence of FR1 is shown as SEQ ID NO.27, and specifically comprises the following steps: AVQLVESGGGLVQAGGSLRVSCEASGFLFN; the amino acid sequence of FR2 is shown as SEQ ID NO.28, and specifically comprises the following steps: WYRQVPGRQRELVA; the amino acid sequence of FR3 is shown as SEQ ID NO.29, and specifically comprises the following steps: RFTISRDNAKNTIFLQMSGLKPEDTAVYYCYA; the amino acid sequence of FR4 is shown as SEQ ID NO.10, and specifically comprises the following steps: WGQGTQVTVSS; the amino acid sequence of CDR1 is shown as SEQ ID NO.30, specifically: GDAYVLG; the amino acid sequence of CDR2 is shown in SEQ ID NO.31, specifically: TITSSGDTSYADSVKG; the amino acid sequence of CDR3 is shown in SEQ ID NO.32, specifically: ERWNGLLQR.
In the nano antibody NbN6, the amino acid sequence of FR1 is shown as SEQ ID NO.33, and specifically comprises the following steps: AVQLVESGGGLVQPGGSLRLSCAASG I IFR; the amino acid sequence of FR2 is shown as SEQ ID NO.34, specifically: WYRQAPGKQRELVA; the amino acid sequence of FR3 is shown as SEQ ID NO.35, and specifically comprises the following steps: RFTISRDNAKNTVYLQMNSLKPEDTGVYYCAA; the amino acid sequence of FR4 is shown as SEQ ID NO.17, and specifically comprises the following steps: WGKGTQVTVSS; the amino acid sequence of CDR1 is shown in SEQ ID NO.36, specifically: FRT I S; the amino acid sequence of CDR2 is shown in SEQ ID NO.37, specifically: SI SGGSSTSYADTVKG; the amino acid sequence of CDR3 is shown in SEQ ID NO.38, specifically: SHTMVGWIYGMDY.
In the nano antibody NbN11, the amino acid sequence of FR1 is shown as SEQ ID NO.39, and specifically comprises the following steps: AVQLVESGGGLVQAGGSLRLSCAASGSFFS; the amino acid sequence of FR2 is shown as SEQ ID NO.34, specifically: WYRQAPGKQRELVA; the amino acid sequence of FR3 is shown as SEQ ID NO.40, and specifically comprises the following steps: RFTISRDNAKNTIYLQMNSLKPEDTAVYYCNA; the amino acid sequence of FR4 is shown as SEQ ID NO.10, and specifically comprises the following steps: WGQGTQVTVSS; the amino acid sequence of CDR1 is shown in SEQ ID NO.41, specifically: INTMG; the amino acid sequence of CDR2 is shown in SEQ ID NO.42, specifically: QITNAGNTNYADSVKG; the amino acid sequence of CDR3 is shown in SEQ ID NO.43, specifically: GQLWGSYRS.
In a second aspect, an embodiment of the present application provides a method for preparing a nanobody targeting a novel coronavirus, including the steps of:
s01, providing target proteins, coating the target proteins on an immune tube, and carrying out enrichment screening to obtain a phage library;
s02, performing second generation sequencing on eluent of the phage library, and synthesizing a gene sequence of the nano antibody with the initial counting front ratio of 10 according to a sequencing result;
s03, cloning the gene sequence of the nano antibody into an expression vector to obtain a recombinant plasmid, transferring the recombinant plasmid into a host cell to induce expression and purifying to obtain the nano antibody of the targeted novel coronavirus.
According to the preparation method of the nano antibody targeting the novel coronavirus Omicron and the subtype BA.2 thereof, which is provided by the second aspect of the embodiment of the application, a nano antibody library is constructed by taking RBD proteins of the novel coronavirus Omicron and the subtype BA.2 thereof as targets based on phage display screening technology, and the nano antibody targeting the novel coronavirus Omicron and the subtype BA.2 thereof is obtained by combining second-generation sequencing, sequence synthesis, cloning, expression and purification; the preparation method utilizes the biological characteristics of phage fast replication in host bacteria, can rapidly screen the nano antibody combined with the specific variant Omikovia and subtype BA.2RBD protein thereof with high flux, is rapid and simple, is favorable for large-scale screening, and improves the screening efficiency.
In step S01, target proteins are provided, and the target proteins are coated on an immune tube for enrichment screening to obtain a phage library.
In some embodiments, the protein of interest is provided as the spinous process protein RBD domain of the novel coronavirus variant, omicron (Omicron), and the concentration of the protein of interest is provided at a concentration of 10-12 μg/mL. In a specific embodiment, the protein of interest is coated onto the immune tube at a concentration of 10. Mu.g/mL.
In some embodiments, in the step of enrichment screening, 2-3 rounds of enrichment screening are performed. Through multiple rounds of enrichment, the obtained library is ensured to have large capacity, and the screening of positive clones is facilitated.
In step S02, the eluent of the phage library is subjected to second generation sequencing, and the gene sequence of the nanobody with the initial pre-count ratio of 10 is synthesized according to the sequencing result.
In the step S03, the gene sequence of the nano antibody is cloned into an expression vector to obtain a recombinant plasmid, and the recombinant plasmid is transferred into a host cell to induce expression and purify to obtain the nano antibody of the targeted novel coronavirus.
In some embodiments, the expression vector is selected from the group consisting of pcoldi vectors, and the gene sequences of the nanobodies are cloned into pcoldi vectors to obtain recombinant plasmids; further, the hemagglutinin tag (hemagglutinin HA tag) was fusion expressed simultaneously during cloning for subsequent detection.
In some embodiments, the host cell is selected from E.coli, and the recombinant plasmid is transferred into E.coli cells to induce expression and purified.
In some embodiments, the recombinant plasmid is transferred into E.coli cells to induce expression and purified as follows: (1) The IPTG with the concentration of 0.2mM is used for induction expression at the low temperature of 16 ℃, so that inclusion bodies and protein degradation can be prevented from being formed; (2) Performing a large amount of induction expression according to the pre-experiment induction conditions, and performing bacteria breaking under the working condition of 1000W of a high-pressure bacteria breaker; (3) Centrifuging at 17000g and 4deg.C for 30min, incubating supernatant with Ni filler at 4deg.C for 1 hr, and eluting; (4) And (3) carrying out molecular sieve separation after Ni column purification, wherein AKATA parameters are set at a flow rate of 0.5 mL/min, and the nano antibody is obtained after collecting once every 1 mL.
In a third aspect, embodiments of the present application disclose the use of nanobodies targeting novel coronaviruses in the preparation of a medicament for the prevention and/or treatment of novel coronavirus pneumonia.
The application of the nanobody of the targeting novel coronavirus Omicron and subtype BA.2 thereof in preparing the medicament for preventing and/or treating the novel coronavirus pneumonia provided by the third aspect of the application is that the nanobody of the targeting novel coronavirus Omicron and subtype BA.2 thereof obtained by screening comprises at least one of nanobody NbN2, nanobody NbN3, nanobody NbN4, nanobody NbN5, nanobody NbN6 and nanobody NbN11, has better neutralization activity capability and inhibition activity capability on viruses of wild strains and variant strains of the novel coronavirus, and is suitable for preparing related medicaments for preventing and/or treating the novel coronavirus pneumonia.
The fourth aspect of the embodiment of the application discloses application of a nanobody targeting a novel coronavirus in preparation of a detection reagent or a detection kit for detecting the novel coronavirus.
The application of the nanobody for targeting the novel coronavirus Omicron and the subtype BA.2 thereof in preparing the detection reagent or the detection kit for detecting the novel coronavirus provided by the fourth aspect of the application provides the nanobody for targeting the novel coronavirus Omicron and the subtype BA.2 thereof, which comprises at least one of the nanobody NbN2, the nanobody NbN3, the nanobody NbN4, the nanobody NbN5, the nanobody NbN6 and the nanobody NbN11, has higher binding capacity for the wild strain of the novel coronavirus and the viruses of various variants, and is suitable for preparing the detection reagent or the detection kit for detecting the novel coronavirus.
The following description is made with reference to specific embodiments.
Example 1
Nano antibody for targeting novel coronavirus Omicron and subtype BA.2 and preparation method thereof
The test steps are as follows:
(1) Coating target protein Omicron RBD on an immune tube according to the concentration of 10 mug/mL, and carrying out 3 rounds of enrichment screening; (2) Second generation sequencing using third round phage eluate to determine sequence information; (3) Designing and synthesizing the screened nano antibody according to the sequencing information; (4) Cloning the nano antibody gene sequence into a pColdII vector, and simultaneously fusion-expressing a hemagglutinin tag (hemagglutinin HA tag) for subsequent detection; (5) Inducing by using IPTG with the concentration of 0.2mM at the low temperature of 16 ℃ (6) carrying out a large amount of induced expression according to the pre-experimental inducing conditions, and carrying out bacteria breaking under the working condition of a high-pressure bacteria breaker of 1000W; (7) 17000g, centrifuging at 4 ℃ for 30min, taking supernatant and incubating with Ni filler at 4 ℃ for 1 hour; (8) And (3) carrying out molecular sieve separation after Ni column purification, wherein AKATA parameters are set at a flow rate of 0.5 mL/min, and collecting once every 1mL to obtain the nanobody targeting the novel coronavirus Omicron and the subtype BA.2 thereof.
Analysis of results:
screening and identification of Omicron RBD nanobodies
Screening of phage nanobody library was performed on omicon RBD, after three rounds of screening, as shown in fig. 1, the library was enriched approximately 3000 times, and the third round phage eluate was subjected to second generation sequencing, as shown in fig. 2, and the sequences with the first 10 content were selected for expression, respectively: nbN2, nbN3, nbN4, nbN5, nbN6, nbN7, nbN8, nbN9, nbN10, nbN11.
Further purifying to obtain 10 high-purity nano antibodies, wherein the protein results of the 10 nano antibodies obtained by purifying are shown in figure 3, and the size of the protein is about 15 kD. And the correctness of the protein is verified by anti-HA WB detection, as shown in figure 4, 10 clones obtained by screening are correctly expressed.
(II) preliminary ELISA identification of nanobody binding to RBD of each variant strain of SARS-CoV-2
The test steps are as follows:
the HA tag is fused into a nanobody gene coding sequence, 10 nanobodies NbN2, nbN3, nbN4, nbN5, nbN6, nbN7, nbN8, nbN9, nbN10 and NbN11 with the HA tag are expressed, an Omicron RBD protein is coated and blocked by ELISA plates, then nanobodies with various concentrations are added for incubation for 1 hour at room temperature, PBS is used for rinsing 3 times, the anti-HA antibody is incubated for 1 hour at room temperature, and then the horseradish peroxidase labeled anti-HA antibody amplifies the signal, TMB color is developed, and meanwhile, the control of irrelevant nanobodies and the blank control of irrelevant protein antigens are made.
Analysis of results
ELISA was performed on 10 nanobodies whose expression was confirmed to be correct by the purification of the above step, and SARS-CoV-2WT, delta (B.1.617.2), omicron (B.1.1.529) RBD protein and bovine serum albumin (Bovine Serum Albumin, BSA), wherein the corresponding legends of 10 nanobodies NbN2, nbN3, nbN4, nbN5, nbN6, nbN7, nbN8, nbN9, nbN10, nbN11 in the figures are as follows: NGS2, NGS3, NGS4, NGS5, NGS6, NGS7, NGS8, NGS9, NGS10, NGS11.
The results of the analysis of the binding to Omicron RBD protein are shown in fig. 5, and it can be seen that, among them, six nanobodies of NbN2, nbN3, nbN4, nbN5, nbN6, and NbN11 showed binding activity to Omicron RBD.
The results of the analysis of binding to SARS-CoV-2 Wild-Type (WT) RBD protein are shown in FIG. 6, wherein six nanobodies of NbN2, nbN3, nbN4, nbN5, nbN6 and NbN11 exhibit binding activity to Omicron RBD.
The results of the analysis of binding to Delta RBD protein are shown in FIG. 7, and it can be seen that, among them, six nanobodies of NbN2, nbN3, nbN4, nbN5, nbN6, nbN11 showed binding activity to Omicron RBD.
The results of the analysis of binding to the negative control protein bovine serum albumin are shown in FIG. 8, and it can be seen that, among them, 10 nanobodies were found to have no binding activity to BSA protein.
Therefore, it was confirmed that 6 nanobodies (Nb N2, nbN3, nbN4, nbN5, nbN6, nbN 11) had binding activity to each variant RBD of SARS-CoV-2, but did not bind to the negative control protein BSA (FIGS. 5-8).
Affinity of six positive nano antibodies with RBD protein of each variant strain SARS-CoV-2
The test steps are as follows:
6 nanobodies (Nb N2, nbN3, nbN4, nbN5, nbN6, nbN 11) were provided, and the affinity of each nanobody to each variant SARS-CoV-2 (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein was analyzed and detected based on the molecular interaction analysis platform Biacore.
Analysis of results
The affinities of the 6 positive nanobodies Nb N2, nbN3, nbN4, nbN5, nbN6 and NbN11 with the RBD proteins of each variant strain (WT, alpha, beta, delta, omicron and Omicron BA.2) of SARS-CoV-2 are shown in Table 1, and the affinities are at the nanomolar level, so that the affinity is higher.
TABLE 1
(IV) 5 positive nanobody and surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
The test steps are as follows:
experiments were used to verify the direct interaction of the in vitro purified nanobody expressed in vitro with the in vitro purified antigen protein and to calculate the equilibrium constants of the two. Multicycle kinetics were run on Biacore S200 to study the binding kinetics of nanobodies and omacron RBD proteins. 5 positive nanobodies Nb D2, nbE8, nbF2, nbG and NbG were provided, and new crown wild strain (WT), variant Alpha (Alpha), beta (Beta), delta (Delta) and Omicron (Omicron) and its subtype BA.2RBD proteins were immobilized on the surface of a CM5 Series S sensor chip at an operating temperature of 25℃respectively, and the nanobodies diluted in a double ratio were flowed over the chip surface at a rate of 30. Mu.L/min for 120 seconds followed by dissociation for 240 seconds to make a kinetic curve, and each relevant parameter was calculated.
Analysis of results
(1) Nb N2 nanobody surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
SPR results showed that Nb N2 nanobody was bound to each variant (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein of SARS-CoV-2, and that Nb N2 had an affinity of 24.45nM to WT RBD protein as shown in FIG. 9; as shown in FIG. 10, nb N2 has an affinity of 43nM with Alpha RBD protein; as shown in FIG. 11, nb N2 has an affinity of 28.87nM for Beta RBD protein; as shown in FIG. 12, nb N2 has an affinity of 97.75nM for the Delta RBD protein; as shown in FIG. 13, nb N2 has an affinity of 55.44nM with Omicron RBD protein; as shown in FIG. 14, nb N2 has an affinity of 8.33nM with the Omacron BA.2RBD protein.
(2) Nb N3 nanobody surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
SPR results showed that Nb N3 nanobody was bound to each variant (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein of SARS-CoV-2, as shown in FIG. 15, the affinity of Nb N3 to WT RBD protein was 12.09nM; as shown in FIG. 16, nb N3 has an affinity of 21.24nM with Alpha RBD protein; as shown in FIG. 17, nb N3 has an affinity of 16.92nM for Beta RBD protein; as shown in FIG. 18, nb N3 has an affinity of 18.65nM for the Delta RBD protein; as shown in FIG. 19, nb N3 has an affinity of 4.79nM with Omicron RBD protein; as shown in FIG. 20, nb N3 had an affinity of 8.77nM for the Omacron BA.2RBD protein.
(3) Nb N4 nanobody surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
SPR results showed that Nb N4 nanobody was bound to each variant (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein of SARS-CoV-2, and that Nb N4 had an affinity of 2.89pM with WT RBD protein as shown in FIG. 21; as shown in FIG. 22, nb N4 has an affinity of 73.09nM for Alpha RBD protein; as shown in FIG. 23, nb N4 has an affinity of 46.4nM for Beta RBD protein; as shown in FIG. 24, nb N4 has an affinity of 23.42nM for the Delta RBD protein; as shown in FIG. 25, nb N4 had an affinity of 5.09nM for Omicron RBD protein; as shown in FIG. 26, nb N4 had an affinity of 38.93nM for the Omacron BA.2RBD protein.
(4) Nb N5 nanobody surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
SPR results showed that Nb N5 nanobody was bound to each variant (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein of SARS-CoV-2, as shown in FIG. 27, nb N5 had an affinity of 0.21nM to WT RBD protein; as shown in FIG. 28, nb N5 has an affinity of 80.48nM for Alpha RBD protein; as shown in FIG. 29, nb N5 has an affinity of 37.22nM for Beta RBD protein; as shown in FIG. 30, nb N5 has an affinity of 24.46nM for the Delta RBD protein; as shown in FIG. 31, nb N5 has an affinity of 90.88nM with Omicron RBD protein; as shown in FIG. 32, nb N5 had an affinity of 20.38nM for the Omacron BA.2RBD protein.
(5) Nb N6 nanobody surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
SPR results showed that Nb N6 nanobody was bound to each variant (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein of SARS-CoV-2, as shown in FIG. 33, nb N6 has an affinity of 38.65nM to WT RBD protein; as shown in FIG. 34, nb N6 has an affinity of 101.3nM with Alpha RBD protein; as shown in FIG. 35, nb N6 has an affinity of 65.36nM for Beta RBD protein; as shown in FIG. 36, nb N6 has an affinity of 26.91nM for the Delta RBD protein; as shown in FIG. 37, nb N6 has an affinity of 78.77nM with Omicron RBD protein; as shown in FIG. 38, nb N6 has an affinity of 89.52nM for the Omacron BA.2RBD protein.
(6) Nb N11 nanobody surface plasmon resonance (Surface Plasmon Resonance, SPR) analysis
SPR results showed that Nb N11 nanobody was bound to each variant (WT, alpha, beta, delta, omicron and Omicron BA.2) RBD protein of SARS-CoV-2, and that Nb N11 had an affinity of 36.06nM to WT RBD protein as shown in FIG. 39; as shown in FIG. 40, nb N11 has an affinity of 26.18nM with Alpha RBD protein; as shown in fig. 41, nb N11 has an affinity 19.17 nM for Beta RBD protein; as shown in FIG. 42, nb N11 has an affinity of 28.66nM for the Delta RBD protein; as shown in FIG. 43, nb N11 has an affinity of 47.39nM for omacron RBD protein; as shown in FIG. 44, nb N11 had an affinity of 105nM for the Omacron BA.2RBD protein.
(fifth) six Positive nanobodies competitive ELISA experiments with ACE2
Test procedure
According to SPR results, selecting nano antibodies with better affinity for competitive ELISA experiment verification. Omicron RBD protein was coated on ELISA plates at a concentration of 0.5. Mu.g/mL overnight at 4 ℃. The PBS was washed once for the next day and blocked with 3% BSA for 2 hours at room temperature. Nanobody fold dilution was performed according to the lowest saturation concentration of ACE2, i.e. the dilution was 0.1% pbst solution containing 0.5 μg/mL ACE2, and incubated for 2.5 hours at room temperature. Rinse with 0.1% pbst.
Analysis of results
As shown in fig. 45, it was examined whether or not these 6 nanobodies compete with ACE2 for binding to omicron RBD protein, nanobody dilution was performed with 0.1% pbst solution containing the lowest saturation concentration of ACE2, and incubated with ELISA plates coated with omicron RBD protein in advance for 2.5 hours at room temperature. 0.1% PBST was rinsed 3 times, and the anti-ACE2 antibody was used to detect the ACE2 content, while a blank control without nanobody and without ACE2 protein was made. The results show that Nb N2 can significantly competitively inhibit ACE2 binding to omicron RBD protein at a half inhibitory concentration (half maximum inhibitory concentration, IC 50) of 0.09 μm. The results show that Nb N2 nanobodies can significantly competitively inhibit ACE2 binding to omicron RBD protein.
Therefore, the nanometer antibody of the novel coronavirus Omicron and subtype BA.2 thereof provided by the application comprises at least one of nanometer antibody NbN2, nanometer antibody NbN3, nanometer antibody NbN4, nanometer antibody NbN5, nanometer antibody NbN6 and nanometer antibody NbN11, and the provided nanometer antibody has higher affinity to RBD domains of novel coronavirus wild strains, novel coronavirus variant Alpha (Alpha), novel coronavirus Beta (Beta), novel coronavirus Delta (Delta), novel coronavirus Omicron and subtype BA.2 thereof, can neutralize different novel coronaviruses rapidly and specifically, and effectively inhibit the activity of the novel coronavirus; meanwhile, the provided nano antibody has a longer CDR3 sequence, is beneficial to flexibly regulating the conformation of the nano antibody, improves the binding effect with viruses, has lower preparation cost and is beneficial to wide application.
The foregoing disclosure is illustrative of the present invention and is not to be construed as limiting the scope of the invention, which is defined by the appended claims.
Sequence listing
<110> Shenzhen City people Hospital
<120> nanobody targeting novel coronavirus, preparation method and application thereof
<140> 2022106375212
<141> 2022-03-31
<160> 49
<170> SIPOSequenceListing 1.0
<210> 1
<211> 128
<212> PRT
<213> Artificial Sequence
<400> 1
Met Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly
1 5 10 15
Gly Ser Leu Arg Leu Ser Cys Ala Ala Thr Gly Leu Thr Phe Thr Gly
20 25 30
Tyr Val Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Asn Arg Glu Phe
35 40 45
Val Ala Ala Val Ser Arg Ser Gly Val Val Thr Arg Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn Thr Val
65 70 75 80
Tyr Leu Gln Met Asn Thr Leu Asn Pro Glu Asp Thr Ala Leu Tyr Tyr
85 90 95
Cys Ala Ala Asp Trp His Pro Leu Thr Gly Ile Met Thr Thr Asp Ser
100 105 110
Thr Glu Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 2
<211> 121
<212> PRT
<213> Artificial Sequence
<400> 2
Met Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly
1 5 10 15
Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Arg
20 25 30
Tyr Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Phe
35 40 45
Val Ala Gly Ile Ser Gly Ser Gly Ile Val Thr Asn Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr
85 90 95
Cys Ala Pro Leu Ile Leu Ala Arg Thr Val Asp Met Asn His Trp Gly
100 105 110
Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 3
<211> 123
<212> PRT
<213> Artificial Sequence
<400> 3
Met Ala Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly
1 5 10 15
Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Gly Phe Ser Leu
20 25 30
Tyr Gly Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu
35 40 45
Val Ala Ser Ile Ala Ser Gly Gly Asp Ser Thr Asn Tyr Gln Asp Ser
50 55 60
Val Lys Gly Arg Phe Ser Met Ser Arg Asp Asn Ala Lys Ser Thr Val
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Val Ala Glu Arg Val Ser Ser Phe Leu Lys Ile Arg Thr Ala Tyr
100 105 110
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 4
<211> 120
<212> PRT
<213> Artificial Sequence
<400> 4
Met Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly
1 5 10 15
Gly Ser Leu Arg Val Ser Cys Glu Ala Ser Gly Phe Leu Phe Asn Gly
20 25 30
Asp Ala Tyr Val Leu Gly Trp Tyr Arg Gln Val Pro Gly Arg Gln Arg
35 40 45
Glu Leu Val Ala Thr Ile Thr Ser Ser Gly Asp Thr Ser Tyr Ala Asp
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr
65 70 75 80
Ile Phe Leu Gln Met Ser Gly Leu Lys Pro Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Tyr Ala Glu Arg Trp Asn Gly Leu Leu Gln Arg Trp Gly Gln
100 105 110
Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 5
<211> 122
<212> PRT
<213> Artificial Sequence
<400> 5
Met Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly
1 5 10 15
Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Ile Phe Arg Phe
20 25 30
Arg Thr Ile Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu
35 40 45
Val Ala Ser Ile Ser Gly Gly Ser Ser Thr Ser Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys
85 90 95
Ala Ala Ser His Thr Met Val Gly Trp Ile Tyr Gly Met Asp Tyr Trp
100 105 110
Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 6
<211> 118
<212> PRT
<213> Artificial Sequence
<400> 6
Met Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly
1 5 10 15
Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Phe Phe Ser Ile
20 25 30
Asn Thr Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu
35 40 45
Val Ala Gln Ile Thr Asn Ala Gly Asn Thr Asn Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Ile Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Asn Ala Gly Gln Leu Trp Gly Ser Tyr Arg Ser Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 7
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 7
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Thr Gly Leu Thr Phe Thr
20 25 30
<210> 8
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 8
Trp Phe Arg Gln Ala Pro Gly Lys Asn Arg Glu Phe Val Ala
1 5 10
<210> 9
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 9
Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Thr Leu Asn Pro Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Ala
20 25 30
<210> 10
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 10
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 11
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 11
Gly Tyr Val Met Ala
1 5
<210> 12
<211> 17
<212> PRT
<213> Artificial Sequence
<400> 12
Ala Val Ser Arg Ser Gly Val Val Thr Arg Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 13
<211> 18
<212> PRT
<213> Artificial Sequence
<400> 13
Asp Trp His Pro Leu Thr Gly Ile Met Thr Thr Asp Ser Thr Glu Tyr
1 5 10 15
Asp Tyr
<210> 14
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 14
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser
20 25 30
<210> 15
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 15
Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Phe Val Ala
1 5 10
<210> 16
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 16
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ala Pro
20 25 30
<210> 17
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 17
Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 18
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 18
Arg Tyr Ala Met Gly
1 5
<210> 19
<211> 17
<212> PRT
<213> Artificial Sequence
<400> 19
Gly Ile Ser Gly Ser Gly Ile Val Thr Asn Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 20
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 20
Leu Ile Leu Ala Arg Thr Val Asp Met Asn His
1 5 10
<210> 21
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 21
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Gly Phe Ser
20 25 30
<210> 22
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 22
Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala
1 5 10
<210> 23
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 23
Arg Phe Ser Met Ser Arg Asp Asn Ala Lys Ser Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Val Ala
20 25 30
<210> 24
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 24
Leu Tyr Gly Met Ser
1 5
<210> 25
<211> 17
<212> PRT
<213> Artificial Sequence
<400> 25
Ser Ile Ala Ser Gly Gly Asp Ser Thr Asn Tyr Gln Asp Ser Val Lys
1 5 10 15
Gly
<210> 26
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 26
Glu Arg Val Ser Ser Phe Leu Lys Ile Arg Thr Ala Tyr
1 5 10
<210> 27
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 27
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Val Ser Cys Glu Ala Ser Gly Phe Leu Phe Asn
20 25 30
<210> 28
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 28
Trp Tyr Arg Gln Val Pro Gly Arg Gln Arg Glu Leu Val Ala
1 5 10
<210> 29
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 29
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Ile Phe Leu Gln
1 5 10 15
Met Ser Gly Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Tyr Ala
20 25 30
<210> 30
<211> 7
<212> PRT
<213> Artificial Sequence
<400> 30
Gly Asp Ala Tyr Val Leu Gly
1 5
<210> 31
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 31
Thr Ile Thr Ser Ser Gly Asp Thr Ser Tyr Ala Asp Ser Val Lys Gly
1 5 10 15
<210> 32
<211> 9
<212> PRT
<213> Artificial Sequence
<400> 32
Glu Arg Trp Asn Gly Leu Leu Gln Arg
1 5
<210> 33
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 33
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Ile Phe Arg
20 25 30
<210> 34
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 34
Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala
1 5 10
<210> 35
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 35
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Ala Ala
20 25 30
<210> 36
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 36
Phe Arg Thr Ile Ser
1 5
<210> 37
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 37
Ser Ile Ser Gly Gly Ser Ser Thr Ser Tyr Ala Asp Thr Val Lys Gly
1 5 10 15
<210> 38
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 38
Ser His Thr Met Val Gly Trp Ile Tyr Gly Met Asp Tyr
1 5 10
<210> 39
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 39
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Phe Phe Ser
20 25 30
<210> 40
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 40
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Ile Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala
20 25 30
<210> 41
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 41
Ile Asn Thr Met Gly
1 5
<210> 42
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 42
Gln Ile Thr Asn Ala Gly Asn Thr Asn Tyr Ala Asp Ser Val Lys Gly
1 5 10 15
<210> 43
<211> 9
<212> PRT
<213> Artificial Sequence
<400> 43
Gly Gln Leu Trp Gly Ser Tyr Arg Ser
1 5
<210> 44
<211> 384
<212> DNA
<213> Artificial Sequence
<400> 44
atggcggtgc agctggtgga gtctggggga ggattggtgc aggctggggg ctctctgaga 60
ctctcctgtg cagccactgg actcaccttc actggttatg taatggcctg gttccgccag 120
gctccaggga agaaccgtga gtttgtagcg gccgttagcc ggagtggtgt tgttacacgc 180
tatgcagact ccgtgaaggg ccgattcacc gtctccagag acaacgccaa gaacacggtg 240
tatctgcaaa tgaacaccct gaatcctgag gacacggccc tttattactg tgcagcagat 300
tggcatccac taactggaat aatgacgact gactctacag agtatgacta ctggggccag 360
gggacccagg tcaccgtctc ctca 384
<210> 45
<211> 363
<212> DNA
<213> Artificial Sequence
<400> 45
atggcggtgc agctggtgga gtctggggga ggattggtgc aggctggggg ctctctgaga 60
ctctcctgtg cagcctctgg acgcaccttc agtaggtatg ccatgggctg gttccgccag 120
gctccaggga aggagcgcca gtttgtagca ggtattagcg ggagtggtat agtcacaaac 180
tatgcagact ccgtgaaggg ccgattcacc atctccagag acaacgccaa gaacacggtg 240
tacctgcaaa tgaacagcct gaaacctgag gacacggcca tttattactg tgcaccgttg 300
atcctagcac gaactgtgga catgaaccac tggggcaaag ggacccaggt caccgtctcc 360
tca 363
<210> 46
<211> 369
<212> DNA
<213> Artificial Sequence
<400> 46
atggcggtgc agctggtgga gtctggggga ggagtggtgc agcctggggg gtctctgaga 60
ctctcctgta cagcctctgg attcggcttc agtctttatg gcatgagctg gtaccgtcag 120
gctccaggga aggagcgcga gttggtcgca tctattgcca gtggtggtga tagtacaaac 180
tatcaagact ccgtgaaggg ccgattctcc atgtccagag acaatgccaa gagcacggtg 240
tatctgcaaa tgaacagcct gaaacctgag gacacggccg tgtattactg cgtggcagaa 300
agagtctcgt cgttcttaaa aattcgtacc gcctactggg gccaggggac ccaggtcacc 360
gtctcctca 369
<210> 47
<211> 360
<212> DNA
<213> Artificial Sequence
<400> 47
atggcggtgc agctggtgga gtctggggga ggcttggtgc aggctggggg gtctctgaga 60
gtgtcctgtg aagcctctgg attccttttc aatggcgatg cctatgtctt gggctggtac 120
cgccaggttc caggtaggca gcgtgaattg gtcgcaacta ttacgagtag cggtgacaca 180
agctatgcgg actccgtgaa gggccgattc accatctcca gagacaacgc caagaataca 240
atttttctgc aaatgagcgg cctgaaaccc gaggacacgg ccgtctatta ctgctatgca 300
gagaggtgga atgggctcct tcaacgctgg ggccagggga cccaggtcac cgtctcctca 360
<210> 48
<211> 366
<212> DNA
<213> Artificial Sequence
<400> 48
atggcggtgc agctggtgga gtctggggga ggcttggtgc aacctggggg gtctctgaga 60
ctctcctgtg cagcctctgg aatcatcttc agattcagaa ccatatcctg gtaccgccag 120
gctccaggga agcagcgcga gttggtcgca agtattagtg gcggcagtag cacgtcctat 180
gcagacactg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acaggcgttt attactgtgc agcgtcacat 300
actatggttg ggtggatcta cggcatggac tactggggca aagggaccca ggtcaccgtc 360
tcctca 366
<210> 49
<211> 354
<212> DNA
<213> Artificial Sequence
<400> 49
atggcggtgc agctggtgga gtctggggga ggcttggtgc aggctggggg gtctctgaga 60
ctctcctgtg cagcctctgg aagcttcttc agtatcaata ccatgggctg gtaccgccag 120
gctccaggga agcagcgcga gttggtcgca caaattacta atgcaggtaa cacaaactat 180
gcagactccg tgaagggccg gttcaccatc tccagagaca acgccaagaa cacgatatat 240
ctgcaaatga acagcctgaa acctgaggac acggccgtct attactgcaa tgcaggacag 300
ttgtggggta gttaccgctc ctggggccag gggacccagg tcaccgtctc ctca 354
Claims (5)
1. The nano antibody for targeting the novel coronavirus is characterized by being a nano antibody NbN6, wherein the amino acid sequence of the nano antibody NbN6 is shown as SEQ ID No. 5.
2. The nanobody of claim 1, wherein said nanobody comprises 4 framework regions FR1, FR2, FR3, FR4 and 3 complementarity determining regions CDR1, CDR2, CDR3;
in the nano antibody NbN6, the amino acid sequence of FR1 is shown as SEQ ID NO.33, the amino acid sequence of FR2 is shown as SEQ ID NO.34, the amino acid sequence of FR3 is shown as SEQ ID NO.35, the amino acid sequence of FR4 is shown as SEQ ID NO.17, the amino acid sequence of CDR1 is shown as SEQ ID NO.36, the amino acid sequence of CDR2 is shown as SEQ ID NO.37, and the amino acid sequence of CDR3 is shown as SEQ ID NO. 38.
3. The nanobody for targeting a novel coronavirus according to claim 1, wherein the base sequence of the nanobody NbN6 is shown as seq id No. 48.
4. Use of the nanobody of any one of claims 1-3 for targeting a novel coronavirus in the preparation of a medicament for preventing and/or treating a novel coronavirus pneumonia.
5. Use of the nanobody of any one of claims 1-3 for targeting a novel coronavirus in the preparation of a detection reagent or a detection kit for detecting a novel coronavirus.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210637521.2A CN115043934B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210637521.2A CN115043934B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210330386.7A CN114478757B (en) | 2022-03-31 | 2022-03-31 | Nano antibody targeting new coronavirus, and preparation method and application thereof |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210330386.7A Division CN114478757B (en) | 2022-03-31 | 2022-03-31 | Nano antibody targeting new coronavirus, and preparation method and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115043934A CN115043934A (en) | 2022-09-13 |
CN115043934B true CN115043934B (en) | 2023-06-27 |
Family
ID=81488870
Family Applications (6)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210637519.5A Active CN115043933B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210637547.7A Active CN115043935B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210637521.2A Active CN115043934B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210638231.XA Active CN115043937B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210330386.7A Active CN114478757B (en) | 2022-03-31 | 2022-03-31 | Nano antibody targeting new coronavirus, and preparation method and application thereof |
CN202210638220.1A Active CN115043936B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210637519.5A Active CN115043933B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210637547.7A Active CN115043935B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
Family Applications After (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210638231.XA Active CN115043937B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
CN202210330386.7A Active CN114478757B (en) | 2022-03-31 | 2022-03-31 | Nano antibody targeting new coronavirus, and preparation method and application thereof |
CN202210638220.1A Active CN115043936B (en) | 2022-03-31 | 2022-03-31 | Nanometer antibody targeting novel coronavirus and preparation method and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (6) | CN115043933B (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114773459B (en) * | 2022-06-16 | 2022-09-09 | 深圳大学 | Nano antibody for resisting novel coronavirus and variant thereof, preparation method and application thereof |
CN114773464B (en) * | 2022-06-20 | 2022-08-26 | 北京市疾病预防控制中心 | Single-domain antibody VHH-2 aiming at new coronavirus omicron strain S protein, coding sequence and application |
CN115772218B (en) * | 2022-11-04 | 2023-08-29 | 四川大学 | Broad-spectrum neutralizing nanobody of two pan-variant strains targeting novel coronavirus S protein receptor binding region and potential application thereof |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6018032A (en) * | 1995-09-11 | 2000-01-25 | Kyowa Hakko Kogyo Co., Ltd. | Antibody against human interleukin-5-receptor α chain |
CN111647076A (en) * | 2020-04-27 | 2020-09-11 | 南京医科大学 | Neutralizing single-domain antibody for resisting novel coronavirus SARS-Cov-2 and application thereof |
CN113754739A (en) * | 2020-06-03 | 2021-12-07 | 中国人民解放军军事科学院军事医学研究院 | Preparation method and application of coronavirus S protein RBD glycoprotein |
Family Cites Families (30)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8907065B2 (en) * | 2006-12-15 | 2014-12-09 | Ablynx N.V. | Polypeptides that modulate the interaction between cells of the immune system |
GB201115214D0 (en) * | 2011-09-02 | 2011-10-19 | Health Prot Agency | Influenza virus antibody compositions |
TR201819577T4 (en) * | 2013-04-29 | 2019-01-21 | Agrosavfe N V | Agrochemical compositions containing antibodies that bind sphingolipids. |
SG10202007233TA (en) * | 2014-04-10 | 2020-09-29 | Lava Therapeutics B V | IMMUNOGLOBULINS BINDING HUMAN Vγ9Vδ2 T CELL RECEPTORS |
CN114106183B (en) * | 2019-01-15 | 2023-06-23 | 浙江道尔生物科技有限公司 | anti-CLD 18A2 nano antibody and application thereof |
CN111116752B (en) * | 2019-12-24 | 2021-09-03 | 北京纽安博生物技术有限公司 | Immunoglobulin-binding single domain antibody, anti-avian influenza single domain antibody, bifunctional antibody and application thereof |
WO2021156490A2 (en) * | 2020-02-06 | 2021-08-12 | Vib Vzw | Corona virus binders |
EP4106806A1 (en) * | 2020-02-21 | 2022-12-28 | Harpoon Therapeutics, Inc. | Flt3 binding proteins and methods of use |
CN111592595B (en) * | 2020-04-27 | 2021-02-19 | 南京医科大学 | Neutralizing antibody against novel coronavirus SARS-Cov-2 and application thereof |
EP4146689A1 (en) * | 2020-05-04 | 2023-03-15 | The Rosalind Franklin Institute | Single domain antibodies binding to sars-cov-2 spike protein |
US20220074938A1 (en) * | 2020-05-07 | 2022-03-10 | Senseutics Limited | Method of detecting pathogens and/or antigens in samples |
AR122104A1 (en) * | 2020-05-15 | 2022-08-10 | Univ Austral De Chile | VHH SIMPLE DOMAIN ANTIBODIES AGAINST SARS-CoV-2 VIRUS AND RAPID METHOD FOR OBTAINING VHH |
WO2022015573A2 (en) * | 2020-07-13 | 2022-01-20 | President And Fellows Of Harvard College | Sars-cov-2 antigen-binding proteins and uses thereof |
WO2022015668A1 (en) * | 2020-07-15 | 2022-01-20 | Regents Of The University Of Minnesota | SARS-CoV-2 NANOBODIES AND METHODS OF USE THEREOF |
WO2022011717A1 (en) * | 2020-07-17 | 2022-01-20 | 上海洛启生物医药技术有限公司 | Nanobody against novel coronavirus, and use thereof |
US20230235027A1 (en) * | 2020-07-23 | 2023-07-27 | The Usa, As Represented By The Secretary, Dept. Of Health And Human Services | Nanobodies directed to coronavirus spike protein receptor binding domain and uses thereof |
KR20220012810A (en) * | 2020-07-23 | 2022-02-04 | (주)셀트리온 | SARS-CoV-2 Virus Neutralizing Binding Molecule Binding To A Epitopes Of Spike Protein Of SARS-CoV-2 Virus |
EP3944877A1 (en) * | 2020-07-29 | 2022-02-02 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Nanobodies sars-cov2 |
WO2022032139A1 (en) * | 2020-08-07 | 2022-02-10 | Sorrento Therapeutics, Inc. | Neutralizing antibodies that bind the sars-cov-2 s protein |
US20240043505A1 (en) * | 2020-08-19 | 2024-02-08 | University Of Pittsburgh-Of The Commonwealth System Of Higher Education | Coronavirus nanobodies and methods for their use and identification |
WO2022040603A2 (en) * | 2020-08-21 | 2022-02-24 | The Rockefeller University | Single-domain antibodies that bind sars-cov-2 |
CN112010967B (en) * | 2020-09-07 | 2021-03-23 | 康维众和(中山)生物药业有限公司 | Novel nano antibody for neutralizing toxicity of coronavirus SARS-Cov-2 and preparation method and application thereof |
EP3970798A1 (en) * | 2020-09-18 | 2022-03-23 | NMI Naturwissenschaftliches und Medizinisches Institut an der Universität Tübingen | Sars-cov-2-nanobodies |
CN112062839A (en) * | 2020-09-22 | 2020-12-11 | 石河子大学 | Nano antibody based on novel coronavirus S protein S1 subunit and application thereof |
CN112062840A (en) * | 2020-09-22 | 2020-12-11 | 石河子大学 | Nano antibody based on novel coronavirus S protein and application thereof |
CN112094342B (en) * | 2020-09-25 | 2022-05-13 | 中国科学技术大学 | Alpaca source nano antibody combined with SARS-CoV-2RBD |
CN113563463B (en) * | 2021-06-11 | 2022-05-17 | 中国医学科学院病原生物学研究所 | Neutralizing nano antibody for resisting novel coronavirus SARS-CoV-2 and application thereof |
CN114031685B (en) * | 2022-01-10 | 2022-03-25 | 中国人民解放军军事科学院军事医学研究院 | Fully human anti-new coronavirus neutralizing antibody ZW2G10 and application |
CN114106166B (en) * | 2022-01-29 | 2022-05-06 | 中国疾病预防控制中心病毒病预防控制所 | Humanized anti-neocoronavirus neutralizing antibody D2 and application thereof |
CN114478758B (en) * | 2022-04-01 | 2022-06-21 | 深圳市人民医院 | Nano antibody of targeting SARS-CoV-2, preparation method and application |
-
2022
- 2022-03-31 CN CN202210637519.5A patent/CN115043933B/en active Active
- 2022-03-31 CN CN202210637547.7A patent/CN115043935B/en active Active
- 2022-03-31 CN CN202210637521.2A patent/CN115043934B/en active Active
- 2022-03-31 CN CN202210638231.XA patent/CN115043937B/en active Active
- 2022-03-31 CN CN202210330386.7A patent/CN114478757B/en active Active
- 2022-03-31 CN CN202210638220.1A patent/CN115043936B/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6018032A (en) * | 1995-09-11 | 2000-01-25 | Kyowa Hakko Kogyo Co., Ltd. | Antibody against human interleukin-5-receptor α chain |
CN111647076A (en) * | 2020-04-27 | 2020-09-11 | 南京医科大学 | Neutralizing single-domain antibody for resisting novel coronavirus SARS-Cov-2 and application thereof |
CN113754739A (en) * | 2020-06-03 | 2021-12-07 | 中国人民解放军军事科学院军事医学研究院 | Preparation method and application of coronavirus S protein RBD glycoprotein |
Non-Patent Citations (2)
Title |
---|
The role of single‑domain antibodies (or nanobodies) in SARS‑CoV‑2 neutralization;Arghavan Zebardast等;《Molecular Biology Reports 》;第49卷;第647-656页 * |
新冠病毒刺突蛋白B细胞线性表位发掘;简春利;《重庆理工大学学报(自然科学)》;第35卷(第1期);第230-235页 * |
Also Published As
Publication number | Publication date |
---|---|
CN114478757B (en) | 2022-07-05 |
CN115043937B (en) | 2023-06-02 |
CN115043933A (en) | 2022-09-13 |
CN115043937A (en) | 2022-09-13 |
CN115043933B (en) | 2023-08-08 |
CN115043936B (en) | 2023-06-27 |
CN115043935A (en) | 2022-09-13 |
CN114478757A (en) | 2022-05-13 |
CN115043934A (en) | 2022-09-13 |
CN115043935B (en) | 2023-06-27 |
CN115043936A (en) | 2022-09-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111303279B (en) | Single-domain antibody for novel coronavirus and application thereof | |
CN115043934B (en) | Nanometer antibody targeting novel coronavirus and preparation method and application thereof | |
CN114478758B (en) | Nano antibody of targeting SARS-CoV-2, preparation method and application | |
CN111690058A (en) | Antibodies with neutralizing activity against coronaviruses and uses thereof | |
JP2021144047A (en) | Antibody-mediated neutralization of chikungunya virus | |
WO2023019724A1 (en) | Monoclonal antibody 35b5, preparation method therefor, and use thereof | |
CN113912710B (en) | Monoclonal antibody for resisting novel coronavirus N protein and application thereof | |
CN113861288B (en) | Novel coronavirus SARS-CoV-2 broad spectrum neutralizing antibody and its use | |
WO2023125520A1 (en) | CAMEL-DERIVED NANOBODY WITH HIGH-AFFINITY FOR α, β, γ AND δ MUTANT STRAINS OF SARS-COV-2 | |
CN116621974A (en) | Novel coronavirus SARS-CoV-2 broad spectrum neutralization nano antibody and application thereof | |
WO2022102744A1 (en) | Antibody against spike protein of sars-cov-2 | |
WO2022179535A1 (en) | Anti-sars-cov-2 nucleocapsid protein monoclonal antibody, and preparation method therefor and use thereof | |
CN115087667B (en) | Antigen binding proteins that specifically bind SARS-CoV-2 | |
CN114478755B (en) | Fully human antibody against novel coronavirus, composition and application thereof | |
WO2019165019A1 (en) | Antibodies to human respiratory syncytial virus protein f pre-fusion conformation and methods of use therefor | |
TWI785583B (en) | Monoclonal antibody against new coronavirus and use thereof | |
WO2021233433A1 (en) | Anti-sars-cov-2 spike protein monoclonal antibody | |
CN114539395A (en) | SARS-CoV-2 wild strain and alpha mutant strain camel source high affinity nano antibody | |
CN114933649A (en) | Anti-varicella-zoster virus antibody and application thereof | |
CN106188285B (en) | A kind of single domain antibody neutralizing xinjiang hemorrhagic fever virus | |
WO2024016842A1 (en) | Anti-respiratory syncytial virus neutralizing antibody and use thereof | |
WO2018175518A1 (en) | Mini-protein immunogens displayng neutralization epitopes for respiratory syncytial virus (rsv) | |
CN114805562B (en) | Anti-novel coronavirus humanized nano antibody and application thereof | |
WO2018140242A1 (en) | Structural basis for antibody cross-neutralization of respiratory syncytial virus and human metapneumovirus | |
WO2023143445A1 (en) | Epitope peptide and antibody for treating hbv infection and related diseases |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |