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A great deal about the molecular processes of virus infection in the host is getting uncovered. Two sequential proteolytic cleavages of viral spike protein by host proteases are prerequisites for the entry of the virus into the host cell. The first cleavage occurs at S1\/S2 site by the furin protease, and the second cleavage at a fusion activation site, the S2\u2032 site, by the TMPRSS2 protease. S2\u2032 cleavage site is present in the S2 domain of spike protein followed by a fusion peptide. Given the S2\u2032 site to be conserved among all the SARS\u2010CoV\u20102 variants, we chose an S2\u2032 epitope encompassing the S2\u2032 cleavage site and generated single\u2010chain antibodies (scFvs) through an exhaustive phage display library screening. Crystal structure of a scFv in complex with S2\u2032 epitope was determined. Incidentally, S2\u2032 epitope in the scFv bound structure adopts an alpha\u2010helical conformation equivalent to the conformation of the epitope in the spike protein. Furthermore, these scFvs can bind to the spike protein expressed either in vitro or on the mammalian cell surface. We illustrate a molecular model based on structural and biochemical insights into the antibody\u2010S2\u2032 epitope interaction emphasizing scFvs mediated blocking of virus entry into the host cell by restricting the access of TMPRSS2 protease and consequently inhibiting the S2\u2032 cleavage competitively.<\/jats:p>","DOI":"10.1002\/pro.4575","type":"journal-article","created":{"date-parts":[[2023,1,24]],"date-time":"2023-01-24T07:46:13Z","timestamp":1674546373000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Epitope\u2010directed <scp>anti\u2010SARS\u2010CoV<\/scp>\u20102 <scp>scFv<\/scp> engineered against the key spike protein region could block membrane fusion"],"prefix":"10.1002","volume":"32","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9565-1302","authenticated-orcid":false,"given":"Deepika","family":"Jaiswal","sequence":"first","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology  New Delhi Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6080-3313","authenticated-orcid":false,"given":"Ujjwal","family":"Kumar","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology  New Delhi Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7292-2720","authenticated-orcid":false,"given":"Vineet","family":"Gaur","sequence":"additional","affiliation":[{"name":"National Institute of Plant Genome Research  New Delhi Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9003-7373","authenticated-orcid":false,"given":"Dinakar M.","family":"Salunke","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology  New Delhi Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,2,14]]},"reference":[{"key":"e_1_2_9_2_1","unstructured":"Anon.The OPLS [optimized potentials for liquid simulations] potential functions for proteins energy minimizations for crystals of cyclic peptides and crambin \u2013 PubMed.1988. 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