Wiley, Proteomics, 23-24(13), p. 3406-3410, 2013
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The increasing availability of antibodies towards human proteins enables broad explorations of the proteomic landscape in cells, tissues and body fluids. This includes assays with antibody suspension bead arrays that generate protein profiles of plasma samples by flow cytometer analysis. However, antibody selectivity is context dependent so it is necessary to corroborate on-target detection over off-target binding. To address this, we describe a concept to directly verify interactions from antibody-coupled beads by analysis of their eluates by Western blots and mass spectrometry. We demonstrate selective antibody binding in complex samples with antibodies towards a set of chosen proteins with different abundance in plasma and serum, and illustrate the need to adjust sample and bead concentrations accordingly. The presented approach will serve as an important tool for resolving differential protein profiles from antibody arrays within plasma biomarker discoveries. This article is protected by copyright. All rights reserved.