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Methods Mol Biol. 2013;1023:137-45. doi: 10.1007/978-1-4614-7209-4_8.

Highly multiplexed antibody suspension bead arrays for plasma protein profiling.

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1
Science for Life Laboratory Stockholm, School of Biotechnology, KTH-Royal Institute of Technology, Stockholm, Sweden.

Abstract

Alongside the increasing availability of affinity reagents, antibody microarrays have become a powerful tool to screen for target proteins in complex samples. Applying directly labeled samples onto arrays instead of using sandwich assays offers an approach to facilitate a systematic, high-throughput, and flexible exploration of protein profiles in body fluids such as serum or plasma. As an alternative to planar arrays, a system based on color-coded beads for the creation of antibody arrays in suspension has become available to offer a microtiter plate-based option for screening larger number of samples with variable sets of capture reagents. A procedure was established for analyzing biotinylated samples without the necessity to remove excess labeling substance. We have shown that this assay system allows detecting proteins down into lower pico-molar and higher pg/ml levels with dynamic ranges over three orders of magnitude. Presently, this workflow enables the profiling of 384 samples for up to 384 proteins per assay.

PMID:
23765623
DOI:
10.1007/978-1-4614-7209-4_8
[Indexed for MEDLINE]
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