Optimization of parameters affecting on CHO cell culture producing recombinant erythropoietin

Autor: Marouf Khalili, Rahman Shokri, Roya Khosravi, Lida Shafiee, Amin Ghafuri-Esfahani, Athar Sharifi, Hooman Kaghazian
Rok vydání: 2020
Předmět:
Zdroj: Preparative Biochemistry & Biotechnology. 50:834-841
ISSN: 1532-2297
1082-6068
DOI: 10.1080/10826068.2020.1753072
Popis: Several factors may affect erythropoietin (EPO) sugar structures including designing cell culture procedure, pH, concentration of additives, dissolved oxygen, and other physicochemical parameters. In this study, we investigated the influence of changes in effective parameters and compounds on the growth rate of Chinese hamster ovary cell (CHO) cells producing recombinant EPO. Cell culture was performed at different temperature, buffering conditions, and varied concentrations of additives such as pyruvic acid, insulin, GlutaMAX, and sodium butyrate. Results indicated that the optimal temperature and pH were 37 °C and 7.2, respectively. Also, optimal concentrations for pyruvic acid, butyrate, glutamate, and insulin were obtained to be 20 mM, 1 mM, 2 mM, and 40 μg/mL, respectively. Then, cell culture was performed in microcarrier-coated spinner flasks under the optimized condition. The results showed recombinant human EPO (rhEPO) production with adequate purity. Optimization of physicochemical conditions and culture media are important factors to improve the quantity and quality of protein products. This study showed that cell growth and recombinant EPO protein production significantly increased under the optimized conditions. The results of this research can also be used in scale-up to increase the efficiency of EPO production.
Databáze: OpenAIRE
Externí odkaz: