Glycoengineered Pichia produced anti-HER2 is comparable to trastuzumab in preclinical study

HER3 intracellular domains play a crucial role in HER3/HER2 dimerization and activation of downstream signaling pathways

“…Antibody against β-actin was from Santa Cruz Biotechnology. HER3mAb was transiently expressed in HEK293 cells based on published amino acid sequences using a mammalian expression vector system and the antibody was purified using protein A affinity resin (Fisher Scientific) as described previously (Rothe et al, 2007;Zhang et al, 2011).…”

Glycoengineered Yeast as an Alternative Monoclonal Antibody Discovery and Production Platform

“…glycoengineered Pichia produced anti-HER2 has demonstrated comparable potencies in receptor inhibition assays in vitro compared to CHO derived trastuzumab. They have very similar HER2 and AKT phosphorylation inhibition and therefore both antibodies inhibit tumor cell proliferation at similar levels Liu et al, 2011;Zhang et al, 2011;Potgieter et al, 2009). …”

“…Humanization of the N-glycosylation pathways of the yeast Pichia pastoris has been achieved by eliminating fungal genes responsible for adding high-mannose and concomitantly reconstituting the canonical human pathway (Hopkins et al, 2011;Li et al, 2006;Liu et al, 2011;Potgieter et al, 2010;Ye et al, 2011b). This result of the engineering is a platform that allows not only yeast-based production of therapeutic glycoproteins with human glycosylation, but also provides a versatile tool for glycosylationbased structure-activity-relationship (SAR) studies to optimize therapeutic proteins for better efficacy, PK and drugability Nett et al, 2012;Zhang et al, 2011). In addition, development of antibody surface display on glycoengineered yeast strain as an antibody discovery tool facilitates the earliest stage of antibody discovery and development in the same expression host which is expected to have a positive impact on cycle times and probabilities of success (Lin et al, 2012).…”

“…Recent preclinical and clinical studies have reported superior efficacy utilizing afucosylated mAbs Junttila et al, 2010;von Horsten et al, 2010;Ward et al, 2011;Wong et al, 2010). Non-mammalian expression hosts have also been utilized for producing afucosylated mAbs including insect cells, plants and yeast (Gasdaska et al, 2007;Barbin et al, 2006;Zhang et al, 2011).…”

“…There are multiple advantages to using yeast as an expression system for therapeutic glycoprotein production, including ease of genetic manipulation, stable expression, rapid cell growth, high yield of secreted protein, low-cost scalable fermentation processes and no risk of human pathogenic virus contamination. However, glycoproteins expressed in wild type yeast generally cannot be used for therapeutic applications due to fungal type highmannose glycans which can result in immunogenicity and poor PK in vivo (Zhang et al, 2011). Humanization of the N-glycosylation pathways of the yeast Pichia pastoris has been achieved by eliminating fungal genes responsible for adding high-mannose and concomitantly reconstituting the canonical human pathway (Hopkins et al, 2011;Li et al, 2006;Liu et al, 2011;Potgieter et al, 2010;Ye et al, 2011b).…”