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.
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Acknowledgements
We like to thank the entire staff of the Human Protein Atlas (HPA) for their tremendous efforts within the Human Protein Atlas project. This work is funded by the PRONOVA project (VINNOVA, Swedish Governmental Agency for Innovation Systems), and by grants from the Knut and Alice Wallenberg Foundation and Science for Life Laboratory, Stockholm.
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Drobin, K., Nilsson, P., Schwenk, J.M. (2013). Highly Multiplexed Antibody Suspension Bead Arrays for Plasma Protein Profiling. In: Bäckvall, H., Lehtiö, J. (eds) The Low Molecular Weight Proteome. Methods in Molecular Biology, vol 1023. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-7209-4_8
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DOI: https://doi.org/10.1007/978-1-4614-7209-4_8
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