I-Corps: Innovation Corps Team: Commercial feasibility of a novel strategy for protein purification development

I-Corps：创新军团团队：蛋白质纯化开发新策略的商业可行性

• 批准号: 1445363
• 负责人: Jannette Carey
• 金额: $ 5万
• 依托单位: Princeton University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1445363&HistoricalAwards=false
• 关键词: Corps; Innovation; Team; Commercial; feasibility

Purification is an essential early step in any studies of protein structure and function. Devising a protein purification strategy is typically the most significant bottleneck in all such studies, because the number of possible ways to purify a protein is essentially infinite. The routes to a suitable solution are entirely empirical and require time-consuming trial-and-error approaches by expert operators with specialized equipment. The most common go-around approach today is to fuse the protein of interest to a tag that facilitates purification, but tags can be difficult to remove and can compromise protein properties. Direct purification of the protein in its native condition is considered the gold standard for structure/function studies. This team has developed a method to circumvent current limitations in devising protein purification strategies that can be developed as a marketable product for non-expert users.The use of tagged or fusion proteins to facilitate purification is a recognized cause of artifacts due to perturbation of protein properties. Devising a purification strategy for untagged proteins presently requires empirical optimization by expert users with specialized equipment. End-users of pure proteins today are not always trained in chromatographic principles that would enable devising optimized purification strategies. Even for proteins with established purification schemes, minor mutational changes commonly used to probe protein function may so drastically alter properties as to make previous purification strategies useless. The proposed approach for devising optimized protein strategies for purification of non-tagged proteins represents the transfer of knowledge from basic research on protein purification and characterization to practical applications in science and technology.

纯化是任何蛋白质结构和功能研究中必不可少的早期步骤。在所有这类研究中，设计一种蛋白质纯化策略通常是最重要的瓶颈，因为纯化蛋白质的可能方法基本上是无限的。通向合适解决方案的途径完全是经验的，需要专家操作员用专门的设备进行耗时的反复试验。如今最常见的变通方法是将感兴趣的蛋白质融合到便于纯化的标签上，但标签可能很难去除，而且可能会损害蛋白质的性质。在天然条件下直接纯化蛋白质被认为是结构/功能研究的金标准。该团队已经开发出一种方法来绕过目前在设计蛋白质纯化策略方面的限制，该策略可以开发为非专家用户的市场产品。使用标记的或融合的蛋白质来促进纯化是由于蛋白质性质的扰动而导致的公认的人工制品的原因。目前，设计一种未标记蛋白质的纯化策略需要专家用户使用专门的设备进行经验优化。如今，纯蛋白质的终端用户并不总是接受过色层分析原理的培训，从而能够设计出优化的纯化策略。即使对于已经建立了纯化方案的蛋白质，通常用于探测蛋白质功能的微小突变也可能会如此剧烈地改变蛋白质的性质，以至于以前的纯化策略变得毫无用处。所提出的用于设计优化的蛋白质纯化策略的方法代表了知识从蛋白质纯化和表征的基础研究向科学和技术中的实际应用的转移。