Collaborative Research: Extracellular secretion of affinity-tagged proteins in E. coli and their non-chromatographic purification using intein-mediated removal

合作研究：大肠杆菌中亲和标记蛋白的胞外分泌及其使用内含肽介导的去除的非色谱纯化

• 批准号: 0965973
• 负责人: Ruizhen Chen
• 金额: $ 15万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-15 至 2014-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0965973&HistoricalAwards=false
• 关键词: Collaborative; Research; Extracellular; secretion; affinity

This NSF award by the Chemical and Biological Separations program supports work by Professors Wilfred Chen and Rachel Chen at University of California, Riverside and Georgia Institute of Technology, respectively, to develop an integrated platform for the extracellular secretion of affinity tagged proteins and their non-chromatographic purification. The integration of the development of genetically engineered capturing scaffolds with protein secretion represents a unique effort that expands the fundamental development of protein engineering with the implementation of purification technology.The proposed research will provide a flexible platform for the direct purification of mature authentic proteins from the growth medium while retaining the high specificity of the affinity-based interaction. The integrated platform is expected to simplify and lower the cost of protein production and can be easily adaptable for any affinity tagged protein of interest. This research will combine the expertise of the two PIs in order to provide a proof of concept study for the implementation of this transformative technology for protein purification. Moreover, graduate students participating in this research will gain an integrated perspective of the important interfaces and synergies connecting biochemistry, modern genetics, and process engineering.

这项由化学和生物分离计划颁发的NSF奖支持加州大学河滨分校的Wilfred Chen教授和佐治亚理工学院的Rachel Chen教授分别为亲和标记蛋白质的胞外分泌和非层析纯化开发一个集成平台。将基因工程捕获支架的开发与蛋白质分泌相结合是一项独特的努力，它通过实施纯化技术来扩展蛋白质工程的基础发展。所提出的研究将为从生长介质中直接纯化成熟的真实蛋白质提供一个灵活的平台，同时保留基于亲和力的相互作用的高度特异性。该集成平台有望简化和降低蛋白质生产成本，并可以很容易地适用于任何亲和标记的感兴趣蛋白质。这项研究将结合两个PI的专业知识，为实施这一变革性的蛋白质纯化技术提供概念研究的证明。此外，参与这项研究的研究生将获得连接生物化学、现代遗传学和过程工程的重要接口和协同作用的综合视角。