{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T09:58:02Z","timestamp":1753869482975,"version":"3.41.2"},"reference-count":37,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2024,1,22]],"date-time":"2024-01-22T00:00:00Z","timestamp":1705881600000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82172263","81571979"],"award-info":[{"award-number":["82172263","81571979"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004608","name":"Natural Science Foundation of Jiangsu Province","doi-asserted-by":"publisher","award":["BK20231262"],"award-info":[{"award-number":["BK20231262"]}],"id":[{"id":"10.13039\/501100004608","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2024,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The emergence of rapid and continuous mutations of severe acute respiratory syndrome 2 (SARS\u2010CoV\u20102) spike glycoprotein that increased with the Omicron variant points out the necessity to anticipate such mutations for conceiving specific and adaptable therapies to avoid another pandemic. The crucial target for the antibody treatment and vaccine design is the receptor binding domain (RBD) of the SARS\u2010CoV\u20102 spike. It is also the site where the virus has shown its high ability to mutate and consequently escape immune response. We developed a robust and simple method for generating a large number of functional SARS\u2010CoV\u20102 spike RBD mutants by error\u2010prone PCR and a novel nonreplicative lentivirus\u2010based system. We prepared anti\u2010RBD wild type (WT) polyclonal antibodies and used them to screen and select for mutant libraries that escape inhibition of virion entry into recipient cells expressing human angiotensin\u2010converting enzyme 2 and transmembrane serine protease 2. We isolated, cloned, and sequenced six mutants totally bearing nine mutation sites. Eight mutations were found in successive WT variants, including Omicron and other recombinants, whereas one is novel. These results, together with the detailed functional analyses of two mutants provided the proof of concept for our approach.<\/jats:p>","DOI":"10.1002\/jmv.29425","type":"journal-article","created":{"date-parts":[[2024,1,23]],"date-time":"2024-01-23T06:45:15Z","timestamp":1705992315000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Generation of SARS\u2010CoV\u20102 spike receptor binding domain mutants and functional screening for immune evaders using a novel lentivirus\u2010based system"],"prefix":"10.1002","volume":"96","author":[{"given":"Junli","family":"Jia","sequence":"first","affiliation":[{"name":"Department of Immunology, National Vaccine Innovation Platform, School of Basic Medical Sciences Nanjing Medical University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emanuela","family":"Garbarino","sequence":"additional","affiliation":[{"name":"Department of Immunology, National Vaccine Innovation Platform, School of Basic Medical Sciences Nanjing Medical University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuhang","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Blood Transfusion Children's Hospital of Nanjing Medical University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiaming","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Immunology, National Vaccine Innovation Platform, School of Basic Medical Sciences Nanjing Medical University  Nanjing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Minmin","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Immunology, National Vaccine Innovation Platform, School of Basic Medical Sciences Nanjing Medical University  Nanjing 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