Development and validation of a respiratory syncytial virus multiplex immunoassay

• DOI: 10.1007/s15010-024-02180-6
• Publication Date: 2024-02-08T17:02:19Z
• Authors: Patrick Marsall; Madeleine Fandrich; Johanna Griesbaum; Manuela Harries; Berit Lange; ; Stephanie Ascough; Pete Dayananda; Christopher Chiu; Jonathan Remppis; Tina Ganzenmueller; Hanna Renk; Monika Strengert; Nicole Schneiderh...; Alex Dulovic
• Keywords: Adult; Immunoassay; 0303 health sciences; Research; 610; Infant; Antibodies, Monoclonal; Respiratory Syncytial Virus Infections; Antibodies, Viral; Antibodies, Neutralizing; 03 medical and health sciences; GTP-Binding Proteins; Respiratory Syncytial Virus, Human; 616; Humans; Child; Viral Fusion Proteins

Abstract Purpose Respiratory syncytial virus (RSV) is one of the leading causes of severe respiratory disease in infants and adults. While vaccines and monoclonal therapeutic antibodies either are or will shortly become available, correlates of protection remain unclear. For this purpose, we developed an RSV multiplex immunoassay that analyses antibody titers toward the post-F, Nucleoprotein, and a diverse mix of G proteins.

Methods A bead-based multiplex RSV immunoassay was developed, technically validated to standard FDA bioanalytical guidelines, and clinically validated using samples from human challenge studies. RSV antibody titers were then investigated in children aged under 2 and a population-based cohort.

Results Technical and clinical validation showed outstanding performance, while methodological developments enabled identification of the subtype of previous infections through use of the diverse G proteins for approximately 50% of samples. As a proof of concept to show the suitability of the assay in serosurveillance studies, we then evaluated titer decay and age-dependent antibody responses within population cohorts.

Conclusion Overall, the developed assay shows robust performance, is scalable, provides additional information on infection subtype, and is therefore ideally suited to be used in future population cohort studies.