Oriented Protein Microarray for Functional Studies

用于功能研究的定向蛋白质微阵列

• 批准号: 6833813
• 负责人: Athena Guo
• 金额: $ 36.33万
• 依托单位: MICROSURFACES, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6833813
• 关键词: SDS polyacrylamide gel electrophoresis; biotechnology; enzyme activity; fluorescence microscopy; green fluorescent proteins; immobilized enzymes; metalloendopeptidases; microarray technology; protein quantitation /detection; protein structure function; proteomics; sulfotransferase; surface coating; technology /technique development

DESCRIPTION (provided by applicant): The proposed research is aimed at developing protein microarrays as a superior technology oriented and viable product for the proteomics market. During Phase-I research, the PI and coworkers have successfully demonstrated the feasibility of fabricating protein microarrays with high selectivity and specificity, with uniform and controllable orientation of immobilized protein molecules, and with proteins in native states and easily accessible by targets in the solution. More importantly, preliminary research established not only the advantage of this chemistry in terms of selective protein immobilization without the need of pre-purification but also the necessity of orientational control in ensuring activity of the native state. The latter was demonstrated by the observation that enzymatic activity of oriented sulfotransferases on the surface was the same as that of solution phase enzymes, while randomly oriented enzymes on the surface showed less than 20% of the activity. Under Phase II funding, we propose (1) to establish the generality of oriented proteins on the surface in maintaining native activity; (2) to develop methods for on-chip refolding of immobilized proteins; (3) to further develop surface coating technology for commercialization. The success of Phase-II research will lead to the launch of a superior surface coating technology for protein microarrays with higher performance at potentially lower cost than current technology in the protein microarray market, which is very important for future large scale drug screening and disease diagnosis.

描述（由申请人提供）： 拟议的研究旨在开发蛋白质微阵列，作为蛋白质组学市场的卓越技术和可行的产品。在第一阶段研究中，PI和同事成功证明了具有高选择性和特异性制造蛋白质微阵列的可行性，具有固定蛋白质分子的均匀和可控制的方向，并在本地状态下具有蛋白质，并且在溶液中易于访问。更重要的是，初步研究不仅建立了这种化学的优势，而无需预纯化的选择性蛋白质固定化，而且还需要取得定向控制，以确保天然状态的活性。通过观察结果证明了后者的表面酶活性与溶液相酶的酶活性相同，而表面上随机定向的酶显示出低于活性的20％。在第二阶段的资金下，我们建议（1）在保持天然活性的情况下在表面上建立定向蛋白的一般性； （2）开发用于芯片重折叠固定蛋白的方法； （3）进一步开发表面涂料技术以进行商业化。 II期研究的成功将导致推出一种优质的表面涂料技术，用于蛋白质微阵列，其性能高于蛋白质微阵列市场的当前技术的性能较高，这对于将来的大规模药物筛查和疾病诊断非常重要。