High level expression of recombinant human serum albumin in CHO cells and optimization of its culture technology
10.13200/j.cnki.cjb.003991
- VernacularTitle:重组人血清白蛋白在CHO细胞中的高效表达及其培养工艺优化
- Author:
NIU Yuhui
- Publication Type:Journal Article
- Keywords:
Recombinant human serum albumin;
CHO cells;
High level expression
- From:
Chinese Journal of Biologicals
2023;36(9):1054-1061+1071
- CountryChina
- Language:Chinese
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Abstract:
ObjectiveTo achieve efficient expression of human serum albumin(HSA)in Chinese hamster ovary(CHO)cells and optimize its culture technology,so as to lay a foundation of the large-scale production of HSA.MethodsThe eukaryotic expression vector of HSA was constructed by gene recombination technology,and then electrotransfected into fully suspended CHO cells. The monoclonal cell lines with stable and high expression of HSA were screened by G418 and limited dilution method. By adding glucose,sodium butyrate and supplementalmedium to the basal medium,the cell culture process was optimized to improve the expression of HSA. Finally,the scale-up culture verification was carried out in a 5 L bioreactor.ResultsThe recombinant expression vector pcDNA3.1-HSA was successfully constructed and expressed in fully suspended CHO cells. After two monoclonal screening,the secondary monoclonal cell lines CHO-rHSA-7H2A9 and CHOrHSA-7H2D12 were obtained with high HSA expression of 29. 37 mg/L and 25. 26 mg/L respectively. The HSA expression level reached about 100. 00 mg/L by optimizing the culture process and wasfinally increased to 166. 16 mg/L in the 5 L bioreactor,which was about 30 times higher than that in the supernatant of the first monoclonal cells.Conclusion The high level expression of HSA in CHO cells was achieved,which laid a foundation of the further large-scale production of HSA in the field of biological products and solving the market supply problems.
