Development and validation of multiplex assays for mouse and human IgG and IgA to Neisseria gonorrhoeae antigens

Erica L Stover; Marguerite B Little; Kristie L Connolly; Lixin Li; Robert A Nicholas; Aleksandra E Sikora; Ann E Jerse; Marcia M Hobbs; J Alex Duncan; Andrew N Macintyre

doi:10.1093/infdis/jiae153

Development and validation of multiplex assays for mouse and human IgG and IgA to Neisseria gonorrhoeae antigens

J Infect Dis. 2024 Mar 25:jiae153. doi: 10.1093/infdis/jiae153. Online ahead of print.

Authors

Affiliations

¹ Duke Human Vaccine Institute and Department of Medicine, Duke University School of Medicine, Durham, North Carolina, USA.
² Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
³ Department of Microbiology and Immunology, Uniformed Services University, Maryland, USA.
⁴ Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, Oregon, USA.
⁵ Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
⁶ Vaccine and Gene Therapy Institute, Oregon Health & Science University, Beaverton, Oregon, USA.
⁷ Department of Medicine, Division of Infectious Diseases, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.

PMID: 38526341
DOI: 10.1093/infdis/jiae153

Abstract

There is an urgent need for vaccines against Neisseria gonorrhoeae (Ng), the causative agent of gonorrhea. Vaccination with an outer-membrane vesicle (OMV)-based Neisseria meningitidis (Nm) vaccine provides some protection from Ng; however, the mechanisms underlying this cross-protection are unknown. To address this need, we developed multiplexed bead-based assays for the relative quantification of human and mouse IgG and IgA against Ng antigens. The assays were evaluated for analyte independence, dilutional linearity, specificity, sensitivity, intra- and inter-assay variability, and robustness to sample storage conditions. The assay was then used to test samples from mice and humans immunized with an Nm-OMV vaccine.

Keywords: Neisseria gonorrhoeae; assay development; humoral response monitoring; multiplex bead assay.

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