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Currently, there are no approved treatments or vaccines for ZIKV infection. We describe here the epitopes for six monoclonal antibodies (mAbs) that bind to domain III of the envelope protein of ZIKV, some of which have therapeutic potential. We show that by using hydrogen\u2013deuterium exchange mass spectrometry (HDX\u2010MS), we can identify three spatially distinct epitopes for the six mAbs investigated. The HDX\u2010MS approach identified epitopes for three mAbs that agreed well with previously reported X\u2010ray crystallography data. The HDX\u2010MS determined epitopes for the other three anti\u2010ZIKV mAbs, for which there were no crystal structures, and the epitopes were confirmed by structure\u2010guided mutagenesis and biolayer interferometry competition binding assay. Our results have implications for the design of vaccine and antibody therapeutics against ZIKV and demonstrate the use of HDX\u2010MS as a rapid and valid approach for epitope mapping.<\/jats:p>","DOI":"10.1002\/jms.4685","type":"journal-article","created":{"date-parts":[[2021,1,15]],"date-time":"2021-01-15T18:38:53Z","timestamp":1610735933000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Hydrogen\u2013deuterium exchange mass spectrometry identifies spatially distinct antibody epitopes on domain III of the Zika virus envelope protein"],"prefix":"10.1002","volume":"56","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6295-645X","authenticated-orcid":false,"given":"Jagat","family":"Adhikari","sequence":"first","affiliation":[{"name":"Department of Chemistry Washington University in St. Louis  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haiyan","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Pathology and Immunology Washington University School of Medicine  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Estefania","family":"Fernandez","sequence":"additional","affiliation":[{"name":"Department of Medicine Washington University School of Medicine  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5535-7731","authenticated-orcid":false,"given":"Yining","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Chemistry Washington University in St. Louis  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8791-3165","authenticated-orcid":false,"given":"Michael S.","family":"Diamond","sequence":"additional","affiliation":[{"name":"Department of Pathology and Immunology Washington University School of Medicine  Saint Louis Missouri USA"},{"name":"Department of Medicine Washington University School of Medicine  Saint Louis Missouri USA"},{"name":"Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs Washington University School of Medicine  Saint Louis Missouri USA"},{"name":"Department of Molecular Microbiology Washington University School of Medicine  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8544-2689","authenticated-orcid":false,"given":"Daved H.","family":"Fremont","sequence":"additional","affiliation":[{"name":"Department of Pathology and Immunology Washington University School of Medicine  Saint Louis Missouri USA"},{"name":"Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs Washington University School of Medicine  Saint Louis Missouri USA"},{"name":"Department of Molecular Microbiology Washington University School of Medicine  Saint Louis Missouri USA"},{"name":"Department of Biochemistry and Molecular Biophysics Washington University School of Medicine  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1159-4636","authenticated-orcid":false,"given":"Michael L.","family":"Gross","sequence":"additional","affiliation":[{"name":"Department of Chemistry Washington University in St. Louis  Saint Louis Missouri USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,12,8]]},"reference":[{"key":"e_1_2_5_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00252-16"},{"key":"e_1_2_5_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/v9050097"},{"key":"e_1_2_5_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2017.06.001"},{"key":"e_1_2_5_5_1","doi-asserted-by":"crossref","unstructured":"Oehler E. Watrin L. Larre P. Leparc\u2010Goffart I. Lastere S. Valour F. Baudouin L. Mallet H. Musso D. andGhawche F.(2014)Zika virus infection complicated by Guillain\u2013Barre syndrome\u2014case report French Polynesia December 2013.Euro surveillance: bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin19","DOI":"10.2807\/1560-7917.ES2014.19.9.20720"},{"key":"e_1_2_5_6_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMp1600297"},{"key":"e_1_2_5_7_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1602412"},{"key":"e_1_2_5_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature17994"},{"key":"e_1_2_5_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf5316"},{"key":"e_1_2_5_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2016.04.013"},{"key":"e_1_2_5_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.07.020"},{"key":"e_1_2_5_12_1","first-page":"3","volume-title":"Human Vaccines: Emerging Technologies in Design and Development","author":"Moody LM a 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