Abstract
Comprehensive proteome-scale protein microarrays can be used to simultaneously screen up to several thousand proteins for drug binding, molecular interactions, or enzymatic activity. The ProtoArray ® Human Protein Microarray v4.0 is comprised of over 8,000 human proteins spotted in duplicate on glass slides. The arrayed proteins are expressed as full-length, N-terminal glutathione S-transferase (GST) fusion proteins in a baculovirus system. The proteins are purified under non-denaturing conditions to maximize proper folding and functionality. Examples will be given about how arrays of human proteins have been used to define protein interaction pathways, screen for enzyme substrates, measure enzyme inhibition, and identify protein targets of drugs. In addition, we have demonstrated the utility of these arrays for determining the specificity of antibodies, thus providing a new tool for the development and characterization of therapeutic and diagnostic antibodies. Finally, we will show how these arrays have been used to provide a rapid and sensitive profiling platform to investigate the circulating antibody profile in several disease states.
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