The methylotrophic yeast Pichia pastoris has become an increasingly popular host for recombinant protein expression in recent times. MRL pioneered a glycoengineered humanized P. pastoris expression system that could produce glycoproteins with glycosylation profiles similar to mammalian systems. Therapeutic glycoproteins produced by the humanized P. pastoris platform have shown comparable folding, stability, and in vitro and in vivo efficacies in preclinical models to their counterparts produced from the CHO cells. P. pastoris offers a cost and time efficient alternative platform for therapeutic protein production. This chapter describes a protocol for using P. pastoris to produce full-length monoclonal antibodies. It covers a broad spectrum of antibody expression technologies in P. pastoris, including expression vector construction, yeast transformation, high-throughput strain selection, fermentation, and antibody purification.
Keywords: Fermentation; High-throughput screening; Monoclonal antibody; Pichia pastoris; Purification.