Pichia GlycoSwitch® Protein Expression System
While no single host protein production system produces all types of proteins and biologics, Pichia has become a major platform for protein expression, producing an ever-expanding variety of novel proteins. Pichia protein expression technology has progressed over the last several years to include unique strains that enable Pichia to produce human-like proteins, control glycosylation patterns, create new compositions of matter and deliver the next generation of biologics.
Roland Contreras, Ph.D., Nico Callewaert, Ph.D., and colleagues at Ghent University and the Flanders Institute for Biotechnology (VIB) in Belgium, in collaboration with RCT, have developed a technology that modifies Pichia to make the production of proteins with human-like glycosylation possible. This improvement to RCT's Pichia platform is called Pichia GlycoSwitch®.
The Pichia GlycoSwitch® system consists of patents, strains and vectors that are useful for engineering Pichiato make proteins with a number of different, near-homogenous glycoforms.
Examples of proteins produced in the Pichia GlycoSwitch® include GM-CSF, IL-10 and IL-22.
Figures 1 and 2 illustrate the production and characterization of various glycoforms of IL-10 in Pichia GlycoSwitch® strains. For other examples, see Pieter P Jacobs, Steven Geysens, Wouter Vervecken, Roland Contreras & Nico Callewaert. Nature Protocols (2009) 4, 58-70
RCT is seeking partners to apply and commercialize this Pichia GlycoSwitch® technology.
Figure 1. IL-10 expressed in GS115 and in selected Pichia GlycoSwitch® strains. Protein samples from each indicated culture supernatant were analyzed by SDS-PAGE stained with Coomassie blue. The extensively smeared band of IL-10 produced in the wild type GS115 host results from the extensively mannosylated forms (see glycan analysis of IL-10 in Figure 2).
Figure 2. Glycan analysis of purified IL-10 expressed in GS115 and in selected Pichia GlycoSwitch® strains. N-glycans released from each IL-10 sample purified from the indicated culture supernatant were fluorescently labeling and then analyzed by DSA-FACE. Glucose reference ladder is shown in the top panel.
