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Alliance"],"published-print":{"date-parts":[[2022,6]]},"abstract":"<jats:p>Neutralizing antibodies against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are useful for patients\u2019 treatment of the coronavirus disease 2019 (COVID-19). We report here affinity maturation of monobodies against the SARS-CoV-2 spike protein and their neutralizing activity against SARS-CoV-2 B.1.1 (Pango v.3.1.14) as well as four variants of concern. We selected matured monobodies from libraries with multi-site saturation mutagenesis on the recognition loops through in vitro selection. One clone, the C4-AM2 monobody, showed extremely high affinity (<jats:italic>K<\/jats:italic><jats:sub>D<\/jats:sub> &lt; 0.01 nM) against the receptor-binding domain of the SARS-CoV-2 B.1.1, even in monomer form. Furthermore, the C4-AM2 monobody efficiently neutralized the SARS-CoV-2 B.1.1 (<jats:italic>IC<\/jats:italic><jats:sub>50<\/jats:sub> = 46 pM, 0.62 ng\/ml), and the Alpha (<jats:italic>IC<\/jats:italic><jats:sub>50<\/jats:sub> = 77 pM, 1.0 ng\/ml), Beta (<jats:italic>IC<\/jats:italic><jats:sub>50<\/jats:sub> = 0.54 nM, 7.2 ng\/ml), Gamma (<jats:italic>IC<\/jats:italic><jats:sub>50<\/jats:sub> = 0.55 nM, 7.4 ng\/ml), and Delta (<jats:italic>IC<\/jats:italic><jats:sub>50<\/jats:sub> = 0.59 nM, 8.0 ng\/ml) variants. The obtained monobodies would be useful as neutralizing proteins against current and potentially hazardous future SARS-CoV-2 variants.<\/jats:p>","DOI":"10.26508\/lsa.202101322","type":"journal-article","created":{"date-parts":[[2022,3,7]],"date-time":"2022-03-07T17:00:16Z","timestamp":1646672416000},"page":"e202101322","update-policy":"https:\/\/doi.org\/10.26508\/lsa.crossmarkpolicy","source":"Crossref","is-referenced-by-count":8,"title":["Monobodies with potent neutralizing activity against SARS-CoV-2 Delta and other variants of concern"],"prefix":"10.26508","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2870-6956","authenticated-orcid":true,"given":"Taishi","family":"Kondo","sequence":"first","affiliation":[{"name":"Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya, 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