Song, ShengliManook, MiriamKwun, JeanJackson, Annette MKnechtle, Stuart JKelsoe, Garnett2022-11-012022-11-012021-112399-36422399-3642https://hdl.handle.net/10161/26168Multiplex immunoassays with acellular antigens are well-established based on solid-phase platforms such as the Luminex^® technology. Cell barcoding by amine-reactive fluorescent dyes enables analogous cell-based multiplex assays, but requires multiple labeling reactions and quality checks prior to every assay. Here we describe generation of stable, fluorescent protein-barcoded reporter cell lines suitable for multiplex screening of antibody to membrane proteins. The utility of this cell-based system, with the potential of a 256-plex cell panel, is demonstrated by flow cytometry deconvolution of barcoded cell panels expressing influenza A hemagglutinin trimers, or native human CCR2 or CCR5 multi-span proteins and their epitope-defining mutants. This platform will prove useful for characterizing immunity and discovering antibodies to membrane-associated proteins.AntibodiesCell LineEpitopesFlow CytometryFluorescent DyesHemagglutininsImmunoassayInfluenza A virusMembrane ProteinsMutationProtein MultimerizationReceptors, CCR2Receptors, CCR5Journal article2022-11-01