Glycoprotein Expression in Pichia Pastoris

Figure 1. N-glycosylation pathway in mammals and yeast (The humanization of N-glycosylation pathways in yeast. 2005)

The glycosylation pathways of mammalian and Pichia pastoris began to differ after the formation of core oligosaccharide Man8GlcNA₂. Mammals began to unload mannose residues, while yeast and other fungi added mannose residues. Finally, it is easy to form excessive glycosylation modification with high mannose chain in yeast cells, which will cause an immune response and limit its use as drug protein. Therefore, it is necessary to modify the glycosylation process of Pichia pastoris.

For clients who want to express glycoprotein, Creative BioMart has set up a glycosylation humanization platform based on the Pichia pastoris.

Figure 2. Glycosylation humanization technology based on the Pichia pastoris.(The humanization of N-glycosylation pathways in yeast. 2005)

(1) The activities of several endogenous glycosyltransferases in yeast were eliminated and Mns I was introduced to form Man₅ GlcNAc₂;

(2) GlcNAcTⅠwas introduced and GldAc was added at the end of Man₅ GlcNAc₂;

(3) MnsⅡ was introduced to remove the residual mannose;

(4) Galactose was added to N-glycan containing GlcNAc terminal;

(5) Sialic acid was added to N-glycan containing galactose end;

(6) When exogenous glycosyltransferases were introduced into yeast, these enzymes were properly located.

Technology Features

• Homogeneous glycosylation: produces the same type of glycosylation on each protein.
• High expression: the glycoprotein was expressed at the same high level as Pichia classic.
• Minimize glycosylation abnormalities: eliminate unnecessary glycosylation.
• Custom glycosylation: allows you to customize the amount and type of glycosylation linked to a protein.

Improving the homogeneity of biopharmaceuticals: producing homogeneous glycoproteins with the highest therapeutic benefits.