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The Low Molecular Weight Proteome

Volume 1023 of the series Methods in Molecular Biology pp 137-145

Date:

Highly Multiplexed Antibody Suspension Bead Arrays for Plasma Protein Profiling

  • Kimi DrobinAffiliated withScience for Life Laboratory Stockholm, School of Biotechnology, KTH - Royal Institute of Technology
  • , Peter NilssonAffiliated withScience for Life Laboratory Stockholm, School of Biotechnology, KTH - Royal Institute of Technology
  • , Jochen M. SchwenkAffiliated withScience for Life Laboratory Stockholm, School of Biotechnology, KTH - Royal Institute of Technology

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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.

Key words

Suspension bead array Antibody array Serum Plasma Labeling