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

美国国家科学基金会化学与生物分离项目授予的这一奖项支持了加州大学河滨分校的Wilfred Chen教授和佐治亚理工学院的Rachel Chen教授的工作，他们分别开发了一个用于细胞外分泌亲和标记蛋白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，代表了一种独特的努力，通过纯化技术的实施，扩大了蛋白质工程的基础发展。提出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，同时保留基于亲和相互作用的高特异性。该集成平台有望简化和降低蛋白质生产成本，并且可以很容易地适应任何感兴趣的亲和力标记蛋白质。这项研究将结合两家pi的专业知识，为实施这种蛋白质纯化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一个综合的视角，了解生物化学、现代遗传学和过程工程之间的重要接口和协同作用。

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.