Powering proteomic research with automated capillary immunoassays - quantitative profiling of signaling pathways in limited samples with precision and accuracy

Wednesday, September 24, 2014 — Poster Session IV
10:00 a.m. –12:00 p.m.	FAES Academic Center	NCI	CANCER-6

Authors

• J-Q Chen
• M.A. Herrmann

Abstract

The Simple Western system (ProteinSimple, CA) has been recently developed and implemented in our Collaborative Protein Technology Resource (CPTR) core facility. The system performs automated western analysis in capillaries using nanogram level of proteins, and provides digital data quantitation with good assay sensitivity and reproducibility. It allows multiplex protein analysis with a through-put of 96 sample / target combination in an overnight run. About two hundred assays covering major key signal pathways have been developed / established in our core and successfully applied for high-throughput profiling of cell signaling events. The technology is especially powerful for quantitative proteomic analysis in limited sample sizes, such as stem cells and fine needle aspirates etc. It offers precise and accurate measurement for pharmacodynamic assessment, targeted therapy evaluation and investigation of drug resistance mechanism. The Simple Western system can also perform iso-electric focusing (IEF) immunoassays to separate, detect and quantify different post-translationally modified states of a protein without using modification-specific antibodies. In addition, the IEF analysis provides additional level of information on the activation status of signaling molecules that are not accessible by conventional Western. The potential of using Simple Western system for clinically relevant biomarker development and therapeutic target identification is particularly exciting.

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