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.
