నైరూప్య
Intein-based purification tags in recombinant protein production and new methods for controlling self-cleavage
Changhua Shi, Elif E Miskioglu, Qing Meng & David W Wood*Tag removal, which is often a necessary step for recombinant proteins purified by affinity tags, is usually accomplished using site-specific protease enzymes. Protease costs and the additional separation of the cleaved tag and protease from the final products have hindered the use of this method for many applications. Furthermore, many protease enzymes leave residual amino acids on the final product, which may affect downstream applications. Self-cleaving tags, based on conventional tags combined with various self-cleaving elements, have recently been developed. We will discuss self-cleaving tags based on inteins. Existing commercial and non-commercial intein systems will be discussed, in addition to the limitations of these existent systems and recent efforts to overcome these.