CHO Cells Expressing RhFSH Of Large-scale Serum-free Culture Technology

Chinese hamster ovary cells (CHO) is the most representative of the cells in the animal cell expression system. Used to express the exogenous target protein is up to the most successful class of cells. CHO cells is the preferred system for recombinant glycosylated protein production.Recombinant human follicle-stimulating hormone (Recombinant human follicle-stimulating hormone, rhFSH) is the anterior pituitary basophilic cells synthesize and secrete a glycoprotein gonadotropin, a glycoprotein family.For women, FSH function of ovarian cells and estrogen production must be a growth factor secretion agent and cell growth regulator; For men, its main function is generated and the corpus luteum hormone (LH) control the testiclescombination of factors, in order to activate and to maintain normal sperm number and quality of a class of drugs play an important role in the diagnosis and treatment of infertility.Sale rhFSH are foreign biopharmaceutical company production, if we can produce the same activity and quality of products will have great market prospects.Animal cells, large-scale cultivation process is an important development trend of the current production of the protein products of glycosylation activity. Simplify the production process from the current development trend, the large-scale production, increase the cell density increase per unit of production, reduce production costs is the key to ensuring large-scale production. Cell spread cultivation to achieve suspension of serum-free culture, conclusive evidence of the safety of the cells through the cell identification and amplification through the process of mass culture production. Animal cells, large-scale cultivation bioreactor, the use of batch, continuous, fed-batch type, perfusion type culture to enlarge production. In order to improve batch production in the actual production, reduce production costs, improve product safety, efficacy and quality of controllability, we need to determine appropriate training techniques, and optimized medium, the optimization process additives, most notably for the purpose of product To confirm that the cells can be used for the production of feasibility, safety.In this study, first by rolling the bottles, the small size of the roller bottle culture technology research to determine the feed rate control in20%to50%, cell seeding density of2～5×105cells/ml, stirring speed80～100rpm, to maintain the culture temperature from31to34℃. Followed by different large-scale cultivation of groping through the5L reactor culture, including batch, continuous, fed-batch type, perfusion, and basically decided that the fed-batch cultivation process to better achieve amplification culture. Again detected by engineering CHO cells, confirmed the safety and feasibility of the cells. And then through the study of serum-free formula culture media and additives to further improve the expression level of18mg/L, in order to improve the batch production.Finally, fed-40L reactor Culture Technology stably expressing the target protein rhFSH identified with foreign counterparts have been marketed with the same activity through quality research, and has basically reached the scale of production.