Interlaboratory standardization of the sandwich enzyme-linked immunosorbent assay designed for MATS, a rapid, reproducible method for estimating the strain coverage of investigational vaccines

Brian D. Plikaytis, Maria Stella, Giuseppe Boccadifuoco, Lisa M. DeTora, Mauro Agnusdei, Laura Santini, Brunella Brunelli, Luca Orlandi, Isabella Simmini, Marzia Giuliani, Morgan Ledroit, Eva Hong, Muhamed Kheir Taha, Kim Ellie, Gowrisankar Rajam, George M. Carlone, Heike Claus, Ulrich Vogel, Ray Borrow, Jamie FindlowStefanie Gilchrist, Paola Stefanelli, Cecilia Fazio, Anna Carannante, Jan Oksnes, Elisabeth Fritzsønn, Anne Marie Klem, Dominique A. Caugan, Raquel Abad, Julio A. Vázquez, Rino Rappuoli, Mariagrazia Pizza, John J. Donnelly, Duccio Medini*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    56 Citations (Scopus)

    Abstract

    The meningococcal antigen typing system (MATS) sandwich enzyme-linked immunosorbent assay (ELISA) was designed to measure the immunologic cross-reactivity and quantity of antigens in target strains of a pathogen. It was first used to measure the factor H-binding protein (fHbp), neisserial adhesin A (NadA), and neisserial heparin-binding antigen (NHBA) content of serogroup B meningococcal (MenB) isolates relative to a reference strain, or "relative potency" (RP). With the PorA genotype, the RPs were then used to assess strain coverage by 4CMenB, a multicomponent MenB vaccine. In preliminary studies, MATS accurately predicted killing in the serum bactericidal assay using human complement, an accepted correlate of protection for meningococcal vaccines. A study across seven laboratories assessed the reproducibility of RPs for fHbp, NadA, and NHBA and established qualification parameters for new laboratories. RPs were determined in replicate for 17 MenB reference strains at laboratories A to G. The reproducibility of RPs among laboratories and against consensus values across laboratories was evaluated using a mixed-model analysis of variance. Interlaboratory agreement was very good; the Pearson correlation coefficients, coefficients of accuracy, and concordance correlation coefficients exceeded 99%. The summary measures of reproducibility, expressed as between-laboratory coefficients of variation, were 7.85% (fHbp), 16.51% (NadA), and 12.60% (NHBA). The overall withinlaboratory measures of variation adjusted for strain and laboratory were 19.8% (fHbp), 28.8% (NHBA), and 38.3% (NadA). The MATS ELISA was successfully transferred to six laboratories, and a further laboratory was successfully qualified.

    Original languageEnglish
    Pages (from-to)1609-1617
    Number of pages9
    JournalClinical and Vaccine Immunology
    Volume19
    Issue number10
    DOIs
    Publication statusPublished - Oct 2012

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