Development and Validation of Multiplex Assays for Mouse and Human IgG and IgA to Neisseria gonorrhoeae Antigens

Author:

Stover Erica L1,Little Marguerite B2,Connolly Kristie L3,Li Lixin4,Nicholas Robert A25,Sikora Aleksandra E46,Jerse Ann E3,Hobbs Marcia M7,Duncan J Alex27,Macintyre Andrew N1ORCID

Affiliation:

1. Duke Human Vaccine Institute, Department of Medicine, School of Medicine, Duke University , Durham, North Carolina

2. Department of Pharmacology, School of Medicine, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina

3. Department of Microbiology and Immunology, Uniformed Services University , Bethesda, Maryland

4. Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University , Corvallis, Oregon

5. Department of Microbiology and Immunology, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina

6. Vaccine and Gene Therapy Institute, Oregon Health & Science University , Beaverton, Oregon

7. Division of Infectious Diseases, Department of Medicine, School of Medicine, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina

Abstract

Abstract There is an urgent need for vaccines against Neisseria gonorrhoeae, the causative agent of gonorrhea. Vaccination with an outer membrane vesicle–based Neisseria meningitidis vaccine provides some protection from N. gonorrhoeae; 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 N gonorrhoeae antigens. The assays were evaluated for analyte independence, dilutional linearity, specificity, sensitivity, intra- and interassay variability, and robustness to sample storage conditions. The assay was then used to test samples from mice and humans immunized with an N meningitidis outer membrane vesicle vaccine.

Funder

National Institutes of Health

Publisher

Oxford University Press (OUP)

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