(489c) Engineering Yeast for the Manufacturing of Human Proteins (Faculty Candidate)
AIChE Annual Meeting
2020
2020 Virtual AIChE Annual Meeting
Food, Pharmaceutical & Bioengineering Division
New Frontiers in Biomolecular Engineering
Thursday, November 19, 2020 - 8:30am to 8:45am
We demonstrate three methods to improve the utility of genetic engineering in manufacturing strains of K. phaffii. First, we developed a markerless genome editing tool, eliminating energetically expensive resistance or auxotrophic markers. Second, we applied our engineering tool to humanize the glycosylation in K. phaffii without sacrificing cell viability. We utilize native promoters to precisely control the expression of endogenous and exogenous glycosyltransferases and glycosidases, balancing glycan homogeneity with cell viability. Third, we improve the quality and titer of monoclonal antibody production by introducing human chaperones to assist with protein folding and secretion. Chaperones are essential for antibody production in human cells, but themselves are large, complex molecules that require machinery for efficient folding. We demonstrate that despite adding to the recombinant protein load, chaperones work together with glyco-engineering to enable the production of high quality monoclonal antibodies at industrially relevant titers.