Novel In Vitro Expression Methods for Proteome Analysis

描述(申请人摘要)：蛋白质组学分析是 后基因组时代。虽然人类基因组中的所有基因现在都已经 破译后，这些基因中的大多数功能仍未确定。一种新的 基于In的蛋白质组学分析方法在第一阶段得到了验证 蛋白质的体外表达和分离。到目前为止，这还不可能 为了快速检测和分离在无细胞条件下产生的蛋白质 系统。AmberGen通过开发一套 促进快速非同位素检测的氨基酰化tRNA， 新生蛋白的分离和N-端标记。第二阶段研究将 重点分析体外表达蛋白(live-Pro)来源的文库 从人的cDNA库、mRNA库和质粒库中获得。由这些基因表达的测试蛋白 将分析包括DHFR、RAS、P53和C-Jun在内的文库。质量 光谱分析和毛细管电泳法将作为快速检测方法 筛选tRNA标记的新生蛋白。它的两个具体应用 方法是快速筛选Live-Pro以确定翻译后 修饰和与包括DNA、蛋白酶在内的其他分子的相互作用 和潜在的药物化合物。Live-Pro包括销售的市场机会 试剂、硬件和向制药公司发放许可证的数量估计超过 1亿美元/年。 建议的商业应用：不可用

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