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

0965953Chen化学和生物分离项目授予的NSF奖项支持分别来自加州大学、滨江和格鲁吉亚理工学院的Wilfred Chen和Rachel Chen教授的工作，以开发用于亲和标记蛋白质的细胞外分泌及其非色谱纯化的集成平台。 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，它通过纯化技术的实施扩大了蛋白质工程的基本发展，拟议的研究将为从生长培养基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，并可轻松适用于任何感兴趣的亲和标记蛋白质。本研究将联合收割机结合两名PI的专业知识，为实施这种蛋白质纯化的变革性技术提供概念验证研究。此外，参与这项研究的研究生将获得连接生物化学，现代遗传学和工艺工程的重要接口和协同作用的综合视角。

0965953ChenThis 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.