Strategies for rapid, low-cost biologic production could shorten process development timelines and improve global access to biologic drugs. Pichia pastoris is a promising production host, due to its fast growth rates to high cell density, and high productivity of heterologous proteins with low host cell protein impurities. Most Pichia pastoris processes use the methanol inducible AOX1 promoter, due to its high expression and tight, inducible regulation, which enable rapid outgrowth and more consistent product quality. While these characteristics have resulted in widespread use of this promoter system, using methanol as a carbon source and inducer results in additional metabolic stress and process safety concerns. In this work, we demonstrate improved process productivity with reduced heat generation and oxygen consumption, using genetic engineering and alternative carbon sources to obtain flexible, methanol free process operation. We conducted a broad screen of 30 alternative carbon sources, characterizing growth and productivity of a rotavirus vaccine subunit product, P[8]. RNA-seq was used to better understand how the choice of production carbon source influences gene regulation and specific productivity. The results of these analyses identified a new production carbon source which enables improved volumetric productivity for several protein products, relative to our original methanol-based perfusion process.

Alternative carbon sources enable flexible, methanol-free Pichia pastoris perfusion processes with increased volumetric productivity

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.