Interrogating antiviral antibody responses with multiplexed, high-throughput serum assays

Hornick, A. C.; Walters, L. C.; Dobson, C. S.; Gaglione, S. A.; Birnbaum, M. E.

doi:10.64898/2026.01.03.697501

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bioRxiv · 10.64898/2026.01.03.697501

Interrogating antiviral antibody responses with multiplexed, high-throughput serum assays

Hornick, A. C.·Walters, L. C.·Dobson, C. S.·Gaglione, S. A.·Birnbaum, M. E.

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Abstract

The COVID-19 pandemic underscored the importance of rapidly analyzing antibody responses against emerging viruses. Existing techniques, however, are limited in their ability to probe antibodies recognition of multiple native-conformation antigens simultaneously. To increase the throughput and multiplexability of antibody profiling, we developed Antibody Reactivity Characterization by Antibody-Dependent Enhancement (ARCADE). This assay employs an antigen-agnostic Fc receptor-expressing cell line and a library of antigen-displaying, genetically barcoded lentiviruses that, when mixed with serum, infect cells and integrate their barcodes at rates reflecting the relative abundances and affinities of the antigen-specific antibodies present. Verified using sera from COVID-19-convalescent and - vaccinated donors, ARCADE delivers insights that align with and expand upon those offered by established immunoassays, highlighting, for example, how an mRNA-based vaccine elicits broader and stronger antibody responses than an adenovirus vector-based vaccine. ARCADE can comprehensively assess how infection and vaccination impact antiviral antibody repertoires over time and across patient populations.

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Hornick, A. C., Walters, L. C., Dobson, C. S., Gaglione, S. A., Birnbaum, M. E.. 2026-01-06. Interrogating antiviral antibody responses with multiplexed, high-throughput serum assays. https://doi.org/10.64898/2026.01.03.697501

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