Rapidly bring the
candidates to commercialization
Pichia pastoris expression system is a common foreign protein expression system, which belongs to the same yeast expression system as Saccharomyces cerevisiae. Yeast is a single-cell eukaryotic organism with simple culture conditions, rapid growth and reproduction, and can withstand high hydrostatic pressure, especially suitable for large-scale production of genetic engineering products, which can improve production efficiency and reduce production costs. Yeast expression system has the advantages of both prokaryotic expression system and eukaryotic expression system. It has the function of post-translational modification at the same time of high-level expression of protein, and the genetic stability of foreign protein genes, which is favored by more and more research institutions and pharmaceutical manufacturers. At present, there are pancreatic islet element, hepatitis B surface antigen, human serum albumin, epidermal growth factor, tumor necrosis factor Many protein products such as antibody drugs have been commercially produced using Pichia pastoris expression system.
The advantages of Pichia pastoris expression system are as follows：
It contains a specific AOX (alcohol oxidase gene) strong promoter, and can regulate the expression of foreign genes with methanol;
The expression level is high, and the yield is above 1g/L;
It can be expressed both in cells and out of cells through cell secretion;
For the foreign target gene with the sequence of guiding secretion signal peptide, the target protein can be directly secreted into the fermentation broth, which is conducive to subsequent separation and purification;
The fermentation process is mature and easy to enlarge. There has been a large-scale industrialized and high-density fermentation process, and the cell dry weight is more than 100g/L, and the expression of recombinant protein has been successfully amplified to 10000L;
The fermentation process is mature and easy to enlarge. There has been a large-scale industrialized and high-density fermentation process, and the cell dry weight is more than 100g/L, and the expression of recombinant protein has been successfully amplified to 10000L;
The inheritance of foreign protein gene is stable, and the foreign gene can be integrated into the chromosome of Pichia pastoris, and it will replicate with the chromosome replication, and it is not easy to lose;
As an eukaryotic expression system, Pichia pastoris has the subcellular structure of eukaryotes, and has post-translational modification processing functions such as glycosylation, fatty acylation, protein phosphorylation, etc.
At present, antibody protein drugs are mostly produced by fermentation of CHO cell expression system. Pichia pastoris still has many advantages over CHO cells. First, Pichia pastoris and CHO cells are both eukaryotic cells, which have many advantages of higher eukaryotic expression system, such as protein processing, protein folding and post-translational modification; Secondly, the construction of Pichia pastoris cell lines is faster and does not require time-consuming virus clearance verification steps, which greatly saves the research and development cycle; Finally, the fermentation and culture of Pichia pastoris will be faster, easier, cheaper and more productive than CHO cells. It can be cultivated in a high-density and large-scale manner, which greatly reduces the production cost of antibodies.
Pichia pastoris expression system is one of the most successful foreign protein expression systems at present, belonging to a yeast expression system. Yeast is a single-cell low-grade eukaryotic organism with common culture conditions, rapid growth and reproduction speed, and can withstand high hydrostatic pressure. When used to express genetic engineering products, it can be produced on a large scale, effectively reducing production costs. As a latecomer exogenous protein expression system, yeast expression system is increasingly widely used in the field of genetic engineering because it has the advantages of both prokaryotic and eukaryotic expression systems.
Rocky Biology has successfully established the Pichia pastoris CMC platform, and currently has many complete project development experience. More than 90% of the project output of Pichia pastoris CMC platform is more than 1g/L, and the overall recovery rate of purification platform process is more than 50%. It only takes 3-4 months from cell line construction to protein purification, greatly shortening the project development time.
The small and medium scale expression platform established by Loqi Biology, including 12 7L fermenters and 1 100L fermenter, is equipped with corresponding purification system; In addition, the 500 L Pichia pastoris pilot test GMP clean plant was officially put into operation, which enabled DeLoki Biologics to have the clinical phase I and phase II sample production capacity. The completion of the pilot production workshop of Pichia pastoris nanoantibody 500L GMP fills the industry gap in this field, which is extremely scarce and competitive.
