Novel In Vitro Expression Methods for Proteome Analysis

DESCRIPTION (Applicant's abstract): Proteomic analysis is an important goal for the post-genomic era. While all genes in the human genome have now been decoded, the function of most of these genes remains undetermined. A new approach for proteomic analysis was demonstrated during Phase I based on in vitro expression and isolation of proteins. Until now, it has not been possible to rapidly detect and isolate proteins, which are produced in a cell-free system. AmberGen has solved this problem by developing a set of misaminoacylated tRNAs which facilitate the rapid non-isotopic detection, isolation and N-terminal labeling of nascent proteins. Phase II research will focus on analyzing libraries of in vitro expressed proteins (LIVE-PRO) derived from human cDNA, mRNA and plasmid libraries. Test proteins expressed from these libraries including DHFR, RAS, P53 and C-Jun will be analyzed. Mass spectrometry and capillary electrophoresis will be used as methods for rapidly screening tRNA labeled nascent proteins. Two specific applications of this approach are the rapid screening of LIVE-PRO to determine post-translational modifications and interactions with other molecules including DNA, proteases and potential drug compounds. Market opportunities for LIVE-PRO including sale of reagents, hardware and licenses to drug companies are estimated to exceed $100 million/year. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE