{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T09:26:21Z","timestamp":1776763581010,"version":"3.51.2"},"reference-count":34,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T00:00:00Z","timestamp":1666742400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31770838"],"award-info":[{"award-number":["31770838"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32071256"],"award-info":[{"award-number":["32071256"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008048","name":"Nanjing University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008048","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2022,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Severe fever with thrombocytopenia syndrome (SFTS) is an acute infectious disease caused by novel bunyavirus (SFTSV), with a mortality rate of 6.3%\u2009~\u200930%. To date, there is no specific treatment for SFTS. Previously, we demonstrated that SFTSV surface glycoprotein (Glycoprotein N, Gn) was a potential target for the development of SFTS vaccine or therapeutic antibodies, and anti\u2010Gn neutralizing antibodies played a protective role in SFTS infection. Compared with traditional antibodies, nanobodies from camelids have various advantages, including small molecular weight, high affinity, low immunogenicity, convenient production by gene engineering, <jats:italic>etc<\/jats:italic>. In this study, we combined next\u2010generation sequencing (NGS) with proteomics technology based on affinity purification\u2010mass spectrometry (AP\u2010MS) and bioinformatics analysis to high\u2010throughput screen monoclonal anti\u2010Gn nanobodies from camel immunized with Gn protein. We identified 19 anti\u2010Gn monoclonal nanobody sequences, of which six sequences were selected for recombinant protein expression and purification. Among these six anti\u2010Gn nanobodies, nanobody 57,493 was validated to be highly specific for Gn. The innovative high\u2010throughput technical route developed in this study could also be expanded to the production of nanobodies specific for other viruses like SARS\u2010CoV\u20102.<\/jats:p>","DOI":"10.1002\/pro.4461","type":"journal-article","created":{"date-parts":[[2022,9,30]],"date-time":"2022-09-30T07:53:16Z","timestamp":1664524396000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Development of functional <scp>anti\u2010Gn<\/scp> nanobodies specific for <scp>SFTSV<\/scp> based on next\u2010generation sequencing and proteomics"],"prefix":"10.1002","volume":"31","author":[{"given":"Binghao","family":"Wang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bilian","family":"Huang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2607-4644","authenticated-orcid":false,"given":"Xinyu","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guantong","family":"Qi","sequence":"additional","affiliation":[{"name":"School of Life Science Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yibing","family":"Ding","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haidong","family":"Gao","sequence":"additional","affiliation":[{"name":"Genepioneer Biotechnologies Co. Ltd.  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingzi","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xilin","family":"Wu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Fang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Key Laboratory of Molecular Medicine, Chemistry and Biomedicine Innovation Center Medical School of Nanjing University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2022,10,26]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.201500458"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(15)00177-2"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268815002678"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cir776"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid1905.120245"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.7774\/cevr.2018.7.1.43"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2014.12.002"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05746-9"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20101860"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(81)90082-9"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.13.4.602-614.2000"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/256495a0"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2007.10.011"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2006.02.040"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2167"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-063011-092449"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2008.10.299"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.drudis.2016.04.003"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molimm.2018.02.012"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.519"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2017.01603"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2018.1556465"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1172\/jci.insight.136855"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142735.im0217s103"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1883"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2008.150"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2020.105645"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.4161\/mabs.3.1.14169"},{"key":"e_1_2_9_30_1","first-page":"23","article-title":"The present and future of de novo whole\u2010genome assembly","volume":"19","author":"Sohn JI","year":"2018","journal-title":"Brief Bioinform"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-019-01633-4"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2014.039"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3170"},{"key":"e_1_2_9_34_1","first-page":"D1145","article-title":"The ProteomeXchange consortium in 2020: enabling 'big data' approaches in proteomics","volume":"48","author":"Deutsch EW","year":"2020","journal-title":"Nucleic Acids Res"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab1038"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.4461","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/pro.4461","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.4461","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,22]],"date-time":"2023-08-22T02:31:45Z","timestamp":1692671505000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.4461"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,26]]},"references-count":34,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2022,11]]}},"alternative-id":["10.1002\/pro.4461"],"URL":"https:\/\/doi.org\/10.1002\/pro.4461","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,26]]},"assertion":[{"value":"2022-04-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-09-24","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-10-26","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e4461"}}