{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:49:38Z","timestamp":1777423778991,"version":"3.51.4"},"reference-count":52,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,5,2]],"date-time":"2023-05-02T00:00:00Z","timestamp":1682985600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"Chan Zuckerberg Biohub\u2014San Francisco","doi-asserted-by":"publisher","award":["3R01AI141003-03S1"],"award-info":[{"award-number":["3R01AI141003-03S1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Knight-Hennessy Scholarship","award":["3R01AI141003-03S1"],"award-info":[{"award-number":["3R01AI141003-03S1"]}]},{"name":"Bio-X Stanford Interdisciplinary Graduate Fellowship","award":["3R01AI141003-03S1"],"award-info":[{"award-number":["3R01AI141003-03S1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>A multiplexed enzyme-linked immunosorbent assay (ELISA) that simultaneously measures antibody binding to multiple antigens can extend the impact of serosurveillance studies, particularly if the assay approaches the simplicity, robustness, and accuracy of a conventional single-antigen ELISA. Here, we report on the development of multiSero, an open-source multiplex ELISA platform for measuring antibody responses to viral infection. Our assay consists of three parts: (1) an ELISA against an array of proteins in a 96-well format; (2) automated imaging of each well of the ELISA array using an open-source plate reader; and (3) automated measurement of optical densities for each protein within the array using an open-source analysis pipeline. We validated the platform by comparing antibody binding to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) antigens in 217 human sera samples, showing high sensitivity (0.978), specificity (0.977), positive predictive value (0.978), and negative predictive value (0.977) for classifying seropositivity, a high correlation of multiSero determined antibody titers with commercially available SARS-CoV-2 antibody tests, and antigen-specific changes in antibody titer dynamics upon vaccination. The open-source format and accessibility of our multiSero platform can contribute to the adoption of multiplexed ELISA arrays for serosurveillance studies, for SARS-CoV-2 and other pathogens of significance.<\/jats:p>","DOI":"10.3390\/pathogens12050671","type":"journal-article","created":{"date-parts":[[2023,5,3]],"date-time":"2023-05-03T01:36:38Z","timestamp":1683077798000},"page":"671","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["MultiSero: An Open-Source Multiplex-ELISA Platform for Measuring Antibody Responses to Infection"],"prefix":"10.3390","volume":"12","author":[{"given":"Janie R.","family":"Byrum","sequence":"first","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6930-9645","authenticated-orcid":false,"given":"Eric","family":"Waltari","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Owen","family":"Janson","sequence":"additional","affiliation":[{"name":"Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA"},{"name":"EPPIcenter Program, University of California, San Francisco, CA 94143, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Syuan-Ming","family":"Guo","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jenny","family":"Folkesson","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg 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L.","family":"Forst","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"},{"name":"Department of Applied Physics, Stanford University, Stanford, CA 94305, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongquan","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Electrical Engineering, Stanford University, Stanford, CA 94305, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adam G.","family":"Larson","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Stanford University, Stanford, CA 94305, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3197-8453","authenticated-orcid":false,"given":"Lena","family":"Blackmon","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, 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Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebecca","family":"Hoh","sequence":"additional","affiliation":[{"name":"Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7616-0321","authenticated-orcid":false,"given":"J. Daniel","family":"Kelly","sequence":"additional","affiliation":[{"name":"Department of Epidemiology and Biostatistics, University of California, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey N.","family":"Martin","sequence":"additional","affiliation":[{"name":"Department of Epidemiology and Biostatistics, University of California, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael J.","family":"Peluso","sequence":"additional","affiliation":[{"name":"Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timothy J.","family":"Henrich","sequence":"additional","affiliation":[{"name":"Division of Experimental Medicine, University of California, San Francisco, CA 94110, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steven G.","family":"Deeks","sequence":"additional","affiliation":[{"name":"Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manu","family":"Prakash","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"},{"name":"Department of Bioengineering, Stanford University, Stanford, CA 94305, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bryan","family":"Greenhouse","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"},{"name":"Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA"},{"name":"EPPIcenter Program, University of California, San Francisco, CA 94143, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shalin B.","family":"Mehta","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John E.","family":"Pak","sequence":"additional","affiliation":[{"name":"Chan Zuckerberg Biohub\u2014San Francisco, San Francisco, CA 94158, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,5,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/S0140-6736(20)31483-5","article-title":"Prevalence of SARS-CoV-2 in Spain (ENE-COVID): A Nationwide, Population-Based Seroepidemiological Study","volume":"396","author":"Oteo","year":"2020","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3566","DOI":"10.1038\/s41467-021-23651-6","article-title":"Citywide serosurveillance of the initial SARS-CoV-2 outbreak in San Francisco using electronic health records","volume":"12","author":"Routledge","year":"2021","journal-title":"Nat. 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