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Steven Pelech

Steven Pelech

Kinexus Bioinformatics Corporation, Canada

Title: Tracking Protein Expression, Modifications and Interactions with Antibody Microarrays

Biography

Steven Pelech is the founder, president and CSO of Kinexus Bioinformatics. He was previously the founder and president of Kinetek Pharmaceuticals. His post-doctoral training was with Sir Philip Cohen at the Univ. of Dundee and Nobel laureate Dr. Edwin Krebs at the Univ. of Washington in Seattle. Since 1988, he has concurrently been on faculty at the Univ. of British Columbia (UBC) and is a full professor in the Department of Medicine. He received his B.Sc. (1979; Honours) and Ph.D. (1982) degrees in Biochemistry from UBC. He has authored over 220 peer-reviewed publications about cell signalling and is one of the discoverers of the MAP kinases.

Abstract

High content antibody microarrays such as the Kinex™ KAM-900 series chips permit highly sensitive and semi-quantitative analyses of the expressions, post-translational modifications and interactions of proteins with sub-milligram amounts of lysate proteins recovered from cells and tissues. The KAM antibody microarrays utilize at least 878 pan- and/or phosphosite-specific antibodies for profiling protein kinases, phosphatases and other low abundance cell signaling proteins. Kinexus has developed multiple complementary detection strategies with the KAM chips that enable high depth monitoring of protein levels, phosphorylation and interactions. One method (KAM) involves the capture of in vitro dye-labeled proteins in lysates obtain from cells subjected to diverse treatments. Another method (PAM) involves the detection of general changes in total protein phosphorylation with the pIMAGO stain. A third approach with a dye-labeled reporter antibody in a sandwich antibody microarray format (SAM), for example with our generic phosphotyrosine-specific PYK rabbit polyclonal antibody, allows detection of specific covalent modifications. The SAM method also allows exploration of the interactions of adapter, scaffolding and chaperone proteins with hundreds of potential target signal transduction proteins with dye-labeled reporter antibodies for these highly interactive proteins. Finally, a fourth technique (FAM) utilizes a biotinylated ATP analogue to probe for the accessibility of the ATP binding state in captured protein kinases on KAM chips as a means to determine their possible states of activation. Combining these methods permits a multi-dimension interrogation of the signalling proteomes of cells and tissues and identifies their most dramatic perturbations in response to diverse treatments and pathological conditions.