Fluorescent Protein Targets for CE-LIF Drug Screening

说明(申请人提供)：更快速、更经济的用药方法 _ 制药行业寻求筛查是因为戏剧性的 预计“可下药”目标的数量将会增加。该项目旨在 开发一种基于新型tRNA荧光标记的方法 结合亲和毛细管电泳法测定蛋白质N-末端 (A)。AmberGen将评估一系列单功能和双功能 检测/亲和标签，这些标签将在 蛋白质在无细胞表达过程中的N-末端 制备了tRNA偶联物。因为能够均匀地发出荧光 标记靶蛋白的同时保持其活性，潜在的药物 可以使用ACE与光诱导荧光相结合的方法来筛选相互作用 检测(LIF)。在这种方法中，移动性方面的小变化可以 使用低剂量试剂检测。可光切割的亲和标记提供了 从无细胞裂解物中快速分离表达蛋白质的方法。在……里面 此外，一系列的“化学可寻址”部分将合并在 翻译后结合为分子的蛋白质的N-末端 不能被天然的蛋白质起始系统所适应。在第二阶段， 基于无细胞N末端标记的ACE-LIF药物筛选技术 进行优化。Aclara生物科学公司将协助这项技术的转让 到微流控设备。 建议的商业应用： 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包括药物筛选试剂和硬件在内的商业产品，以及将向制药行业销售的集成系统。

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.