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

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

Pathogens. 2023 May 2;12(5):671. doi: 10.3390/pathogens12050671.

Authors

Janie R Byrum¹, Eric Waltari¹, Owen Janson^{2

3}, Syuan-Ming Guo¹, Jenny Folkesson¹, Bryant B Chhun¹, Joanna Vinden⁴, Ivan E Ivanov¹, Marcus L Forst^{1

5}, Hongquan Li⁶, Adam G Larson⁷, Lena Blackmon¹, Ziwen Liu¹, Wesley Wu¹, Vida Ahyong¹, Cristina M Tato¹, Krista M McCutcheon¹, Rebecca Hoh², J Daniel Kelly⁸, Jeffrey N Martin⁸, Michael J Peluso², Timothy J Henrich⁹, Steven G Deeks², Manu Prakash^{1

7}, Bryan Greenhouse^{1

2

3}, Shalin B Mehta¹, John E Pak¹

Affiliations

¹ Chan Zuckerberg Biohub-San Francisco, San Francisco, CA 94158, USA.
² Division of HIV, Infectious Disease, and Global Medicine, University of California, San Francisco, CA 94143, USA.
³ EPPIcenter Program, University of California, San Francisco, CA 94143, USA.
⁴ Infectious Diseases and Immunity Graduate Program, University of California, Berkeley, CA 94720-3370, USA.
⁵ Department of Applied Physics, Stanford University, Stanford, CA 94305, USA.
⁶ Department of Electrical Engineering, Stanford University, Stanford, CA 94305, USA.
⁷ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
⁸ Department of Epidemiology and Biostatistics, University of California, San Francisco, CA 94158, USA.
⁹ Division of Experimental Medicine, University of California, San Francisco, CA 94110, USA.

Abstract

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.

Keywords: ELISA; SARS-CoV-2; multiplex; open-source; serology; serosurveillance.

Abstract

Grants and funding