Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection

The emergence of COVID-19 necessitated the rapid development of vaccines. While highly effective at reducing severe disease and death, breakthrough infections remain a problem as the virus continues to mutate. To help address this issue, we show the utility of a multiplex immunoassay in measuring mu...

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Main Authors: Troy Odo, Brien K. Haun, Caitlin A. Williams, Aquena Ball, Albert To, Teri Ann S. Wong, Lauren Ching, Eileen Nakano, Alex Van Ry, Laurent Pessaint, Hanne Andersen, Oreola Donini, Vivek R. Nerurkar, Axel T. Lehrer
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Language:English
Published: MDPI AG 2025-03-01
Series:COVID
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Online Access:https://www.mdpi.com/2673-8112/5/4/44
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author Troy Odo
Brien K. Haun
Caitlin A. Williams
Aquena Ball
Albert To
Teri Ann S. Wong
Lauren Ching
Eileen Nakano
Alex Van Ry
Laurent Pessaint
Hanne Andersen
Oreola Donini
Vivek R. Nerurkar
Axel T. Lehrer
author_facet Troy Odo
Brien K. Haun
Caitlin A. Williams
Aquena Ball
Albert To
Teri Ann S. Wong
Lauren Ching
Eileen Nakano
Alex Van Ry
Laurent Pessaint
Hanne Andersen
Oreola Donini
Vivek R. Nerurkar
Axel T. Lehrer
author_sort Troy Odo
collection DOAJ
description The emergence of COVID-19 necessitated the rapid development of vaccines. While highly effective at reducing severe disease and death, breakthrough infections remain a problem as the virus continues to mutate. To help address this issue, we show the utility of a multiplex immunoassay in measuring multiple aspects of the antibody response generated by SARS-CoV-2 vaccines. We use a multiplex immunoassay platform to measure spike-specific IgG concentration, avidity, and receptor-binding inhibition. In addition, we correlate results from an ACE-2 receptor-binding inhibition assay with corresponding data from a SARS-CoV-2 microneutralization assay to establish this inhibitory assay as a potential predictor of virus neutralization. We studied these antibody responses in SARS-CoV-2-naïve and -convalescent vaccinees. Our results showed increased IgG concentrations, avidity, and inhibition following vaccination in both groups. We were also able to differentiate the immune response between the two groups using the multiplex immunoassay platform to look at antibody diversity. The receptor-binding inhibition assay has strong correlations with a cell-based pseudovirus neutralization assay as well as with WT SARS-CoV-2 Washington and Delta variant PRNT<sub>50</sub> assays. This suggests that the inhibition assay may be able to simultaneously predict virus neutralization of different SARS-CoV-2 variants. Overall, we show that the developed custom multiplex immunoassay with several experimental variations is a powerful tool in assessing multiple aspects of the SARS-CoV-2 antibody response in vaccinated individuals.
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spelling doaj-art-0fce203e1a0e4d94850bb34f7d19ec0b2025-08-20T03:13:51ZengMDPI AGCOVID2673-81122025-03-01544410.3390/covid5040044Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior InfectionTroy Odo0Brien K. Haun1Caitlin A. Williams2Aquena Ball3Albert To4Teri Ann S. Wong5Lauren Ching6Eileen Nakano7Alex Van Ry8Laurent Pessaint9Hanne Andersen10Oreola Donini11Vivek R. Nerurkar12Axel T. Lehrer13Department of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USABioqual Inc., Rockville, MD 20850, USABioqual Inc., Rockville, MD 20850, USABioqual Inc., Rockville, MD 20850, USASoligenix, Inc., Princeton, NJ 08540, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USADepartment of Tropical Medicine, Medical Microbiology, and Pharmacology, University of Hawaii Manoa, Honolulu, HI 96813, USAThe emergence of COVID-19 necessitated the rapid development of vaccines. While highly effective at reducing severe disease and death, breakthrough infections remain a problem as the virus continues to mutate. To help address this issue, we show the utility of a multiplex immunoassay in measuring multiple aspects of the antibody response generated by SARS-CoV-2 vaccines. We use a multiplex immunoassay platform to measure spike-specific IgG concentration, avidity, and receptor-binding inhibition. In addition, we correlate results from an ACE-2 receptor-binding inhibition assay with corresponding data from a SARS-CoV-2 microneutralization assay to establish this inhibitory assay as a potential predictor of virus neutralization. We studied these antibody responses in SARS-CoV-2-naïve and -convalescent vaccinees. Our results showed increased IgG concentrations, avidity, and inhibition following vaccination in both groups. We were also able to differentiate the immune response between the two groups using the multiplex immunoassay platform to look at antibody diversity. The receptor-binding inhibition assay has strong correlations with a cell-based pseudovirus neutralization assay as well as with WT SARS-CoV-2 Washington and Delta variant PRNT<sub>50</sub> assays. This suggests that the inhibition assay may be able to simultaneously predict virus neutralization of different SARS-CoV-2 variants. Overall, we show that the developed custom multiplex immunoassay with several experimental variations is a powerful tool in assessing multiple aspects of the SARS-CoV-2 antibody response in vaccinated individuals.https://www.mdpi.com/2673-8112/5/4/44SARS-CoV-2subunit vaccinemRNA vaccinemultiplex immunoassayhumoral immunitymultiplex inhibition test
spellingShingle Troy Odo
Brien K. Haun
Caitlin A. Williams
Aquena Ball
Albert To
Teri Ann S. Wong
Lauren Ching
Eileen Nakano
Alex Van Ry
Laurent Pessaint
Hanne Andersen
Oreola Donini
Vivek R. Nerurkar
Axel T. Lehrer
Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection
COVID
SARS-CoV-2
subunit vaccine
mRNA vaccine
multiplex immunoassay
humoral immunity
multiplex inhibition test
title Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection
title_full Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection
title_fullStr Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection
title_full_unstemmed Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection
title_short Use of a Multiplex Immunoassay Platform to Investigate Multifaceted Antibody Responses in SARS-CoV-2 Vaccinees with and Without Prior Infection
title_sort use of a multiplex immunoassay platform to investigate multifaceted antibody responses in sars cov 2 vaccinees with and without prior infection
topic SARS-CoV-2
subunit vaccine
mRNA vaccine
multiplex immunoassay
humoral immunity
multiplex inhibition test
url https://www.mdpi.com/2673-8112/5/4/44
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