Intensified purification and refolding of inclusion body protein through multi-column simulated moving bed chromatography

点击翻译按钮获取中文摘要

Protein refolding is essential to impart biological activity on proteins which affects therapeutic efficacy. Challenges in protein refolding include low yield, high product dilution, and large process volumes, which translates to exorbitant product costs. Protein purification using single chromatographic columns are commonly used, but disadvantaged by low media utilization, high solvent consumption, and low productivity. Multi-column Continuous Chromatography (MCC) processing based on Simulated Moving Bed (SMB) technology is a promising technology that has had significant impact on pharmaceutical chiral separations, but has not found widespread use in bio-processing due to challenges in processing. This research aims to intensify refolding and purification of few commercially-relevant proteins, for improved process efficiency and productivity via MCC. The new knowledge generated from this fundamental study will address technical challenges that have caused SMB to be largely unexplored for large-scale bio-processing. Particularly, this will enable innovative rational design and scale-up methodology of MCC process aimed to enhance productivity of continuous protein bio-processing. The successful outcome of this work will have broad application for high-throughput of new biologics entering clinical trials reducing drug manufacturing costs and the speed with which new protein drugs can be brought to market. It is expected to have immediate applicability for both new protein-based drugs and "bio-generics", which are competing with drugs coming off-patent on a cost basis. The PI has a unique MCC experimental rig capable be undertaking the proposed research. The requested analytical infrastructure of Waters UPLC H-Class Bio systems will help perform reliable high-resolution rapid analysis of extremely small amounts of target molecules as diverse as intact proteins, protein complexes, and peptides upstream of mass spectrometry in LC-MS workflows. This initiative is unique as there is no similar MCC development on-going anywhere in Canada, and therefore, this investment will be creating a distinctive state-of-the-art research capability on large-scale protein refolding and purification within this country.