With its high sample density and multiplexed experimental settings, protein microarray technology is a powerful tool to study system biology at the whole proteome level. Protein microarray technology has a broad-range of utilities in biomedical research and pharmaceutical development. The most prominent application for this technology is disease biomarker discovery and in vitro diagnosis. The success of this technology fits well with NIH's mission for improving public health and developing novel methods and technologies to address unmet medical needs. Although the development of DNA microarrays has provided the technological platform for high-density microarray fabrication, the greatest challenge in assembling protein microarrays is still the acquisition of high-quality functional proteins encompassing the entire human proteome. The most comprehensive protein microarray chip currently available only includes one third of the human proteome, and furthermore, the proteins on this chip were produced in an insect cell expression system and lack appropriate mammalian posttranslational modifications. The major impediment to the application of protein microarray technology is the lack of a high-quality commercially available chip. OriGene Technologies, Inc. is a gene centric biotech company. To spearhead the human recombinant protein production market, OriGene shuttled more than 15,000 ORF cDNAs into its proprietary Eukaryotic expression system and produced 12,000 C-terminal myc- and Flag-epitope tagged human proteins in a human cell originated HEK293T cell line and validated their expression via an anti-Flag Western blot analysis. This unprecedented work has put OriGene in an enviable position to generate the best human protein microarray chip. Our extensive preliminary work also demonstrated that we will be able to generate the highest quality protein microarray chips for the research reagent market. In this Phase I application, we expect to develop a prototype high density protein microarray chip that contains 10,464 unique recombinant human proteins produced in HEK293T cells. The utilities of this protein chip will be demonstrated by detection of protein-protein interactions. The expected duration of the project is one year. Phase II of the grant will be an expansion into the commercialization of this protein chip technology, and the production of a highly purified protein microarray chip spotted with at least 20,000 unique human proteins. Multiple utilities will also be developed and optimized. We believe that this high density protein microarray chip technology will become a disruptive technology for biomedical research, as well as pharmaceutical research and development. PUBLIC HEALTH RELEVANCE: Protein microarray technology has a broad-range of utilities in biomedical research and pharmaceutical development, including disease biomarker discovery and in vitro diagnosis. The development of this technology fits well with NIH's mission for improving public health and developing novel methods and technologies to address unmet medical needs.