ArticleMethods in molecular biology (Clifton, N.J.)2025
Highly Multiplexed Immunoassays for the Validation of Anti-GPCR Antibodies.
Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Antibodies targeting G protein-coupled receptors (GPCRs) are important tools to study the biology and pharmacology of this important superfamily of cell surface signaling proteins. However, generating anti-GPCR antibodies is challenging, partly due to the high sequence homology of related GPCRs. Immunoassays built on suspension bead arrays (SBAs) can be applied to validate anti-GPCR antibodies for flow-based assay applications by enabling a multi-parallel assessment of on-target and off-target binding. In this chapter, we describe how to immobilize an antibody library to generate SBAs. We further describe how a library of engineered GPCRs can be combined with the multiplexed SBA assay to validate the specificity of the bead-bound anti-GPCR antibodies. The SBA-based approach presented here offers a versatile and robust tool for multiplexed characterization of antibody binding selectivity and off-target interactions, as well as for mapping GPCR epitopes involved in antibody binding.
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