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Selectivity analysis of single binder assays used in plasma protein profiling
Author(s) -
Neiman Maja,
Fredolini Claudia,
Johansson Henrik,
Lehtiö Janne,
Nygren PerÅke,
Uhlén Mathias,
Nilsson Peter,
Schwenk Jochen M.
Publication year - 2013
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201300030
Subject(s) - antibody , context (archaeology) , blot , chemistry , biomarker , computational biology , proteomics , biomarker discovery , chromatography , biology , biochemistry , immunology , paleontology , gene
The increasing availability of antibodies toward 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 W estern blots and MS . We demonstrate selective antibody binding in complex samples with antibodies toward 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.