Fluorescent Protein Targets for CE-LIF Drug Screening

描述（由申请人提供）：更快速、更经济的给药方法 +____________________________________ 筛选是由制药行业寻求由于戏剧性的 预计“可用药”目标的数量会增加。该项目旨在 开发这样一种基于使用新的基于tRNA的荧光标记的方法 结合亲和毛细管电泳技术的蛋白质N-末端 （ACE）。AmberGen将评估一系列单功能和双功能 检测/亲和标签，其将被特异性地并入到 蛋白质在无细胞表达过程中的N-末端， 制备tRNA缀合物。由于能够均匀地发出荧光 标记靶蛋白同时保持其活性，潜在药物 可以使用ACE结合光诱导荧光来筛选相互作用 检测（LIF）。在这种方法中，移动性的小改变可以是 使用低体积试剂检测。可光裂解的亲和标签提供了一种 从无细胞裂解物中快速分离表达蛋白的方法。在 此外，一系列“化学可寻址”部分将在 蛋白质的N-末端，用于翻译后缀合至 不能被天然蛋白质起始系统容纳。在第二阶段， 基于无细胞N-末端标记的ACE-LIF药物筛选技术将 优化。Aclara Biosciences将协助转让这项技术 微流体装置。 拟定商业应用： 基于ACE-LIF的药物筛选无细胞蛋白质标记系统的开发将产生商业产品，包括用于药物筛选的试剂和硬件以及将销售给制药行业的集成系统。

DESCRIPTION (provided by applicant): More rapid and economical methods of drug +____________________________________ screening are sought by the pharmaceutical industry due to the dramatic projected increase in the number of "druggable" targets. This project aims to develop such a method based on the use of novel tRNA-based fluorescent labeling of the N-terminus of proteins combined with affinity capillary electrophoresis (ACE). AmberGen will evaluate a series of monofunctional and bifunctional detection/affinity tags, which will be specifically incorporated at the N-terminus of proteins during their cell-free expression using specially prepared tRNA conjugates. Because of the ability to uniformly fluorescence label target proteins while preserving their activity, potential drug interactions can be screened using ACE combined with light induced fluorescence detection (LIF). In this approach, small alterations in mobility can be detected using low volumes of reagents. Photocleavable affinity tags provide a means to rapidly isolate expressed proteins from the cell-free lysate. In addition, a series of "chemically addressable" moieties will be incorporated at the N-terminus of proteins for post-translational conjugation to molecules that cannot be accommodated by the native protein initiation system. In Phase II, an ACE-LIF drug screening technology based on cell-free N-terminal labeling will be optimized. Aclara Biosciences will assist in the transfer of this technology to microfluidic devices. PROPOSED COMMERCIAL APPLICATION: The development of cell-free protein labeling systems for drug screening based on ACE-LIF will result in commercial products including reagents and hardware for drug screening as well as an integrated system which will be marketed to the pharmaceutical industry.