MultiSero: An Open-Source Multiplex-ELISA Platform for Measuring Antibody Responses to Infection

Janie R. Byrum; Eric Waltari; Owen Janson; Syuan-Ming Guo; Jenny Folkesson; Bryant B. Chhun; Joanna Vinden; Ivan E. Ivanov; Marcus L. Forst; Hongquan Li; Adam G. Larson; Lena Blackmon; Ziwen Liu; Wesley Wu; Vida Ahyong; Cristina M. Tato; Krista M. McCutcheon; Rebecca Hoh; J. Daniel Kelly; Jeffrey N. Martin; Michael J. Peluso; Timothy J. Henrich; Steven G. Deeks; Manu Prakash; Bryan Greenhouse; Shalin B. Mehta; John E. Pak

doi:10.3390/pathogens12050671

Pathogens (May 2023)

MultiSero: An Open-Source Multiplex-ELISA Platform for Measuring Antibody Responses to Infection

Janie R. Byrum,
Eric Waltari,
Owen Janson,
Syuan-Ming Guo,
Jenny Folkesson,
Bryant B. Chhun,
Joanna Vinden,
Ivan E. Ivanov,
Marcus L. Forst,
Hongquan Li,
Adam G. Larson,
Lena Blackmon,
Ziwen Liu,
Wesley Wu,
Vida Ahyong,
Cristina M. Tato,
Krista M. McCutcheon,
Rebecca Hoh,
J. Daniel Kelly,
Jeffrey N. Martin,
Michael J. Peluso,
Timothy J. Henrich,
Steven G. Deeks,
Manu Prakash,
Bryan Greenhouse,
Shalin B. Mehta,
John E. Pak

Affiliations

Janie R. Byrum: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Eric Waltari: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Owen Janson: Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA
Syuan-Ming Guo: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Jenny Folkesson: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Bryant B. Chhun: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Joanna Vinden: Infectious Diseases and Immunity Graduate Program, University of California, Berkeley, CA 94720-3370, USA
Ivan E. Ivanov: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Marcus L. Forst: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Hongquan Li: Department of Electrical Engineering, Stanford University, Stanford, CA 94305, USA
Adam G. Larson: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA
Lena Blackmon: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Ziwen Liu: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Wesley Wu: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Vida Ahyong: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Cristina M. Tato: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Krista M. McCutcheon: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Rebecca Hoh: Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA
J. Daniel Kelly: Department of Epidemiology and Biostatistics, University of California, San Francisco, CA 94158, USA
Jeffrey N. Martin: Department of Epidemiology and Biostatistics, University of California, San Francisco, CA 94158, USA
Michael J. Peluso: Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA
Timothy J. Henrich: Division of Experimental Medicine, University of California, San Francisco, CA 94110, USA
Steven G. Deeks: Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA
Manu Prakash: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Bryan Greenhouse: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
Shalin B. Mehta: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA
John E. Pak: Chan Zuckerberg Biohub—San Francisco, San Francisco, CA 94158, USA

DOI: https://doi.org/10.3390/pathogens12050671
Journal volume & issue: Vol. 12, no. 5
p. 671

Abstract

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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.

Published in Pathogens

ISSN: 2076-0817 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Medicine
Website: http://www.mdpi.com/journal/pathogens

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