Self-Assembling Protein Microchips

描述(申请人提供)：蛋白质生物芯片可以呈现许多 不同的蛋白质同时将极大地加速研究和 医学和生物科学的发展。一个必不可少的要素 生产蛋白质生物芯片是一种快速、灵活的固定化方法 许多不同的蛋白质。采用连续固定化策略的方法 使用机械定位设备会遇到许多问题，并且 他们生产独一无二的芯片的能力受到限制。我们有 完成并行固定化战略的概念验证演示 用于使用自组装标签生产蛋白质生物芯片。蛋白质是 通过体外翻译准备的，这有助于繁重的任务 提纯成千上万种不同的蛋白质。体外翻译技术 这里描述的产生具有标签粘贴在随机位置的蛋白质， 因此，由此产生的固定化蛋白集合呈现随机分布。 方向。这极大地提高了对蛋白质活性的询问。 原则性实验证明了该方案的有效性。 用于蛋白质的特定固定化和检测。该计划的目标是 本提案中描述的工作是扩展这些初步实验和 为了证明多个蛋白质可以被翻译，在空间上多路传输 通过在生物芯片上自组装，并用于功能分析。自动化是 简单易行，并将使生物芯片在理论上具有无限数量 广泛用于商业用途的功能蛋白质。拟议的研究 涉及到一种方法的开发，该方法选择性地绑定十个(作为 可行性论证)不同的合成蛋白质对多个 CombiMatrix生物芯片的预定位置。之前开发的 Combimatix生物芯片是一种可寻址微电极阵列，它允许 特定电极位置上各种预定寡核苷酸的合成 通过一系列的电化学反应。通过添加所需的标签 含寡核苷酸的蛋白质(VIA，例如链霉亲和素/生物素)这种蛋白质可以 附着在一个电极位置，用来合成 互补寡核苷酸。结果是一种生物芯片，它有各种各样的 已知的蛋白质固定在已知的阵列位置。 建议的商业应用： 这项研究旨在开发一种快速、高效、自动化和廉价的定制蛋白质芯片制造方法。这些芯片可用于诊断分析、药物-铅筛选和功能基因组学研究，包括蛋白质-蛋白质和蛋白质-靶标相互作用、催化和抑制活性。

DESCRIPTION (provided by the applicant): Protein biochips that can present many different proteins simultaneously will greatly accelerate research and development in medical and biological sciences. One element essential for producing protein biochips is a fast and flexible method for immobilization of many different proteins. Methods that employ serial immobilization strategies using mechanical spotting devices suffer from numerous problematic issues and are constrained in their ability to produce one-of-a-kind chips. We have completed proof-of-concept demonstrations of a parallel immobilization strategy for producing protein biochips using self-assembling tags. Proteins are prepared by in vitro translation, which facilitates the laborious task of purifying thousands of different proteins. The in vitro translation techniques described here produce proteins with tags affixed at random positions, therefore, the resulting ensemble of immobilized proteins presents random orientations. This greatly improves interrogating the activity of the proteins. Proof-of-principle experiments have demonstrated that this scheme is effective for specific immobilization and detection of proteins. The objective of the work described in this proposal is to extend these preliminary experiments and to demonstrate that multiple proteins can be translated, spatially multiplexed by self-assembly on a biochip, and used for functional analysis. Automation is facile and will enable biochips with theoretically unlimited numbers of functional proteins for extensive commercial use. The proposed research concerns the development of a method to selectively bind ten (as a demonstration of feasibility) different synthetic proteins to multiple predetermined sites of a CombiMatrix biochip. The previously developed Combimatix biochip is an addressable microelectrode array which, allows the synthesis of various predetermined oligonucleotides at specific electrode sites through a sequence of electrochemical/chemical reactions. By tagging desired proteins with oligonucleotides (via, eg. Streptavidin/Biotin) the protein can be attached at an electrode site, which was used to synthesize the complementary oligonucleotide. The result is a biochip, which has a variety of known proteins immobilized in known array positions. PROPOSED COMMERCIAL APPLICATION: The proposed research is directed to develop a method of manufacture of custom protein microchips in a fast, efficient, automatic and inexpensive way. These chips can be used for diagnostic analysis, drug-lead screening, and functional genomics studies including protein-protein and protein-target interactions, catalytic and inhibitory activity.