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

在任何蛋白质结构和功能的研究中，纯化都是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。找到合适的解决方案完全是经验性的，需要专家操作人员用专门的设备进行耗时的反复试验。目前最常见的方法是将感兴趣的蛋白质融合到一个便于纯化的标签上，但是标签很难去除，并且会损害蛋白质的特性。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规避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业用户的可销售产品。使用标记或融合蛋白来促进纯化是由于蛋白质性质的扰动而产生伪影的公认原因。设计一种纯化策略，为未标记的蛋白质目前需要经验优化的专家用户与专门的设备。今天纯蛋白质的最终用户并不总是受过色谱原理的培训，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变化也可能极大地改变蛋白质的性质，从而使以前的纯化策略失效。本文提出的设计非标记蛋白质纯化优化蛋白质策略的方法，代表了知识从蛋白质纯化和表征的基础研究转移到科学和技术的实际应用。

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.