The lack of a platform technology for the purification of non-mAb proteins remains a major concern in the biopharmaceutical industry.  The use of purification tags is ubiquitous at small scale; however, the cost of tag removal becomes prohibitive upon scale-up.  To address this, we have developed self-cleaving tags based on intein technology.  While more cost-effective, non-ideal intein cleaving kinetics currently represent a significant barrier to implementation of this system in the biopharmaceutical industry.The current work describes the development of inteins with optimized cleaving kinetics via directed evolution and yeast surface display.  We report progress toward the development of inteins with a shifted pH optimum for cleaving and increased sensitivity to small molecule inhibition that can be used in mammalian expression systems. Coupling of these novel inteins with a choline-binding domain affinity tag produces a robust purification platform for the production of non-mAb proteins at industrial scale.

Development of a purification platform for non-antibody products via optimized intein self-cleaving tags

245th National Meeting (2013)

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