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Selectivity analysis of single binder assays used in plasma protein profiling
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-1855-703X
KTH, Centres, Science for Life Laboratory, SciLifeLab.
Karolinska Institutet, Solna, Sweden .
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2013 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 13, no 23-24, 3406-3410 p.Article in journal (Refereed) Published
Abstract [en]

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 Western 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.

Place, publisher, year, edition, pages
2013. Vol. 13, no 23-24, 3406-3410 p.
Keyword [en]
Antibodies, Plasma profiling, Protein arrays, Suspension bead arrays, Verification
National Category
Medical Biotechnology
URN: urn:nbn:se:kth:diva-139859DOI: 10.1002/pmic.201300030ISI: 000329994000005ScopusID: 2-s2.0-84890306435OAI: diva2:690153
Science for Life Laboratory - a national resource center for high-throughput molecular bioscienceVinnovaKnut and Alice Wallenberg Foundation

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Available from: 2014-01-22 Created: 2014-01-15 Last updated: 2014-02-14Bibliographically approved

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Neiman, MajaFredolini, ClaudiaLehtiö, JanneNygren, Per-ÅkeUhlén, MathiasNilsson, PeterSchwenk, Jochen M.
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