{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T05:35:15Z","timestamp":1776749715063,"version":"3.51.2"},"reference-count":51,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2025,9,18]],"date-time":"2025-09-18T00:00:00Z","timestamp":1758153600000},"content-version":"vor","delay-in-days":18,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"Beijing Science and Technology New Star Program","award":["Z211100002121005"],"award-info":[{"award-number":["Z211100002121005"]}]},{"name":"Beijing Municipal Science & Technology Plan","award":["Z221100007922016"],"award-info":[{"award-number":["Z221100007922016"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31971377"],"award-info":[{"award-number":["31971377"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2021YFA0910604"],"award-info":[{"award-number":["2021YFA0910604"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,8,31]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Recently, nanobody-based therapeutics have emerged as a highly effective strategy for COVID-19 treatment. However, camelid-derived nanobodies often require humanization engineering to reduce immunogenicity in clinical applications while simultaneously preserving their target-binding affinities. Here, we employed a computational and engineering approach to optimize the binding affinities of complementarity-determining region (CDR)-grafted humanized variants of the camelid-derived nanobody Nb2\u201367, which exhibits potent SARS-CoV-2 neutralization. By grafting the three CDR loops of Nb2\u201367 onto the humanized scaffold of the approved therapeutic nanobody Caplacizumab and refining the target-binding interface, we generated five nanobody variants with improved computational humanness scores. Three of these variants (Nb491, Nb273, and Nb1052) retained neutralizing activity. To further enhance their potency, we fused these variants to a self-assembling scaffold, generating three multivalent constructs with higher humanness scores. Pseudovirus assays showed that all the trivalent nanobodies exhibited picomolar neutralizing potency comparable to the original trivalent Nb2\u201367. Our study presents a novel computational and multivalent engineering strategy that effectively restores the antiviral efficacy of humanized CDR-grafted nanobody variants, offering a valuable approach for developing nanobody-based therapeutics against COVID-19 and other diseases.<\/jats:p>","DOI":"10.1093\/bib\/bbaf477","type":"journal-article","created":{"date-parts":[[2025,8,28]],"date-time":"2025-08-28T11:47:31Z","timestamp":1756381651000},"source":"Crossref","is-referenced-by-count":1,"title":["Computational refinement and multivalent engineering of complementarity-determining region-grafted nanobodies on a humanized scaffold for retaining antiviral efficacy"],"prefix":"10.1093","volume":"26","author":[{"given":"Liyun","family":"Huo","sequence":"first","affiliation":[{"name":"State Key Laboratory of Genetics and Development of Complex Phenotypes, Shanghai Engineering Research Center of Industrial Microorganisms, MOE Engineering Research Center of Gene Technology, School of Life Sciences, Fudan University , Shanghai 200438 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qin","family":"Qin","sequence":"additional","affiliation":[{"name":"Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, MOE Frontiers Center for Brain Science, Fudan University , Shanghai 200032 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tian","family":"Tian","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Genetics and Development of Complex Phenotypes, Shanghai Engineering Research Center of Industrial Microorganisms, MOE Engineering Research Center of Gene Technology, School of Life Sciences, Fudan University , Shanghai 200438 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xing","family":"Zhang","sequence":"additional","affiliation":[{"name":"ACROBiosystems Inc. , Beijing 100176 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoming","family":"He","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Genetics and Development of Complex Phenotypes, Shanghai Engineering Research Center of Industrial Microorganisms, MOE Engineering Research Center of Gene Technology, School of Life Sciences, Fudan University , Shanghai 200438 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuhui","family":"Cao","sequence":"additional","affiliation":[{"name":"ACROBiosystems Inc. , Beijing 100176 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tianfu","family":"Zhang","sequence":"additional","affiliation":[{"name":"ACROBiosystems Inc. , Beijing 100176 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanqin","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Genetics and Development of Complex Phenotypes, Shanghai Engineering Research Center of Industrial Microorganisms, MOE Engineering Research Center of Gene Technology, School of Life Sciences, Fudan University , Shanghai 200438 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5238-1704","authenticated-orcid":false,"given":"Qiang","family":"Huang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Genetics and Development of Complex Phenotypes, Shanghai Engineering Research Center of Industrial Microorganisms, MOE Engineering Research Center of Gene Technology, School of Life Sciences, Fudan University , Shanghai 200438 ,","place":["China"]},{"name":"Multiscale Research Institute of Complex Systems, Fudan University , Shanghai 201203 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