Producing human IgG Fc antibodies in yeasts - Methanol-free expression in Pichia pastoris

Nowadays, therapeutic monoclonal antibodies (mAb) are predominantly produced with mammalian cell culture systems such as those using Chinese hamster ovary (CHO) cells. Efforts are underway to reduce the costs of this process to meet the increasing global demand in biopharmaceuticals; meanwhile cheaper and faster expression systems are being investigated as alternatives. The yeast Pichia pastoris has become a substantial workhorse for recombinant protein production. However, the N-linked glycosylation in P. pastoris, namely high mannose glycosylation, is significantly different from that in CHO or other mammalian cells including human cells. In this webinar, we will show, how we constructed a SuperMan5 strain of P. pastoris using the Pichia GlycoSwitch^® technology to successfully produce a more mammalian-like immunoglobulin fragment IgG Fc. This showcases the potential of P. pastoris as a next generation mAb production platform. Importantly, we will highlight the benefits of working with a methanol-independent promoter. Most P. pastoris promoters used for protein expression are derived from genes in the methanol metabolism pathway, creating safety concerns due to the flammable nature of methanol especially at large scale. Here, a fed-batch SuperMan5 P. pastoris fermentation was carried out in which methanol induction, as well as its affiliated safety risks, was eliminated. The 3 L bench scale bioprocess demonstrates the feasibility of using P. pastoris for IgG Fc production in a methanol-independent environment, providing insight for future industrial mAb production seeking a safe and cost-effective approach to compete with mammalian cell culture.

 

Learning Objectives