With many years of experience in glycoengineering, CD BioGlyco has solved various shortcomings of the expression systems and brought customers high-quality cell lines to produce glycoproteins. We have confidence to be your essential research assistant in the field of glycobiology.

Background

The Pichia pastoris expression system has the characteristics of fast growth, easy culture, simple genetic manipulation, and can perform glycosylation of proteins, It has been recognized as a powerful tool for large-scale protein expression and is broadly applied in bioengineering. Yeast first generates Man₉GlcNAc₂ sugar chain under the action of 1,6 mannose transferase encoded by och1 gene, which triggers the continuous addition of multiple mannoses on the sugar chain, forming a high mannose chain Man_10-100GlcNAc₂ which is different from mammalian cells.

The high mannose chain severely affects the structure and reduces the biological activity of the protein. At the same time, the protein containing the high mannose chain can cause an immune response in the body. Because the yeast expression system has the disadvantage of excessive glycosylation modification, it cannot replace the mammalian cell expression system in the production of various glycoproteins. Therefore, solving the problem of the high mannose chain is the core step of P. pastoris glycosylation engineering.

The therapeutic glycoprotein produced by the current glycoengineered P. pastoris platform has shown the same folding, stability and efficacy in vitro and in vivo as its counterpart produced from CHO cells in preclinical models. It is evident that P. pastoris provides a cost-effective alternative platform for the production of therapeutic glycoproteins.

Fig 1. Overview of yeast and mammalian N-linked glycosylation and the strategy used for glyco-engineered P. pastoris expression system (Vervecken, W.; et al. 2014)

Services

CD BioGlyco applies genetic engineering technology to modify the glycosylation pathway of P. pastoris. By knocking out the och1 gene and introducing mannosidase, we constructed a glycoengineered P. pastoris expression system, which overcomes the excessive glycosylation and the final glycoprotein produced has a uniform low-molecular sugar chain. This new expression system can produce glycoproteins with glycosylation properties similar to mammalian systems. It has the advantages of short production cycle, low cost, high yield, easy operation and conducive to industrialized scale-up production, and will have a significant impact on protein expression and biopharmaceuticals.

The services we provide include but are not limited to the following processes

• Design the target protein gene sequence, add tags, optimize codons and synthesize genes
• Clone the target protein gene into the P. pastoris expression vector
• Transform the constructed expression vector into the glycoengineered P. pastoris strain
• Large-volume cultivation and induce expression of the target protein
• Purify and detect the concentration and purity of the target protein
• Repack or freeze-dry the target protein according to customer requirements

Applications

• Therapeutic glycoprotein production, commercial product development
• Biopharmaceutical research
• Improve the biological activity of recombinant proteins
• Genetic engineering drug production
• Glycoprotein structure and function research

Advantages of Us

• Short production cycle
• Efficient expression vector, optimized protein expression strategies
• One-stop service, guarantee protein quantity and purity
• High stability and consistency of experimental results

CD BioGlyco provides glyco-engineered Pichia pastoris express system that can be used for large-scale industrial production and one-stop services from gene synthesis to protein purification.

Customers can contact our employees directly and we will respond promptly. If you are interested in our services, please contact us for more detailed information.

References:

1. Nylen, A.; Chen, M.T. Production of full-length antibody by Pichia pastoris. Recombinant Glycoprotein Production. 2017.
2. Vervecken, W.; et al. In vivo synthesis of mammalian-like, hybrid-type N-glycans in Pichia pastoris. Applied and environmental microbiology. 2004, 70(5): 2639-2646.