Effects of Glucose and Sodium Butyrate on the rHSA Yield in CHO-rHSA Engineering Cell Line

doi:10.13560/j.cnki.biotech.bull.1985.2021-1238

Abstract

Abstract:

At present,suspension CHO cells are widely used as the most potential engineered cell lines for exogenous proteins. The culture conditions of promoting its high density growth and high expression of exogenous protein have also become the research hotspot. Glucose and sodium butyrate play an important role in promoting the growth of CHO cells and the expression of exogenous proteins. Therefore,study the effect of glucose and sodium butyrate on the expression of recombinant serum albumin in CHO-rHSA engineered cells is of great value. Firstly,the effect of glucose and sodium butyrate on the expression of recombinant serum albumin in CHO-rHSA engineering cells was determined by single factor test,and the optimal addition conditions were selected. Then the combination culture conditions of glucose and sodium butyrate which effectively increased the expression of rHSA were optimized by the two-factor combination experiment. The yield of rHSA increased 121.93% by adding 7 g/L glucose every 48 h from the 3rd day of cell culture,and 110.01% by adding 1.0 mmoL/L sodium butyrate on the 2nd day of cell culture. The combination of sodium butyrate and glucose prolonged the culture time from 7 d to 14 d and increased the expression of rHSA to 85.642 mg/L,which was 212.49% higher than that of untreated cells,134.85% higher than glucose alone,and 88.35% higher than sodium butyrate alone. The culture condition of CHO engineering cell line was determined to add 1.0 mmol/L sodium butyrate and 3.0→7.0 g/L glucose when cells cultured for 48 h. Sodium butyrate combined with glucose can promote foreign protein expression more efficiently,and glucose can weaken the inhibition of sodium butyrate on the growth of CHO cells,which provides a new theoretical basis for enhancing the expression of foreign protein in CHO engineering cells.

Key words: recombinant human serum albumin,rHSA, NaBu, glucose, CHO-rHSA engineering cell line

NIU Yu-hui, LI Xiang-rong, WU Bei, LI Hong-shan, LI Dian-yu, CHEN Lei, WEI Suo-cheng, FENG Ruo-fei. Effects of Glucose and Sodium Butyrate on the rHSA Yield in CHO-rHSA Engineering Cell Line[J]. Biotechnology Bulletin, 2022, 38(7): 278-286.

Figures/Tables 5

Table 1 Different strategies for supplementing glucose

补糖策略 Glucose supplement strategies/（g·L^-1）	补糖前葡萄糖含量 Glucose content in the medium before supplementing glucose/（g·L^-1）	补糖后葡萄糖含量 Glucose content in the medium after supplementing glucose /（g·L^-1）
2.0→5.0	2.0	5.0
2.0→7.0	2.0	7.0
3.0→5.0	3.0	5.0
3.0→7.0	3.0	7.0

Fig.1 Effects of different glucose supplementation strategies on the growth of CHO-rHSA cells and the yield of rHSA A：Viable cell density. B：Viability. C：Glucose concentration. D：The productivity of rHSA was detected by ELISA（**P<0.01）

Fig.2 Effects of different concentrations of sodium butyrate on the growth of CHO-rHSA cells and the yield of rHSA A：Viable cell density. B：Viability. C：Glucose concentration. D：The protein expression of rHSA was detected by Western blot（Sd：Standard concentration 100 mg/L）. E：The relative expression level of rHSA was detected by gray analysis. F：The productivity of rHSA was detected by ELISA（*P<0.05）

Fig.3 Effects of combined culture of glucose and sodium butyrate on the growth of CHO-rHSA cells and the yield of rHSA A：Viable cell density（①：3.0→5.0 g/L. ②：3.0→7.0 g/L）. B：Viability. C：Glucose concentration. D：The protein expression of rHSA was detected by Western blot（Sd：standard concentration 100 mg/L）. E：The relative expression level of rHSA was detected by gray analysis. F：The productivity of rHSA was detected by ELISA（***P<0.001）

Fig.4 Effect of adding time of sodium butyrate on the growth of CHO-rHSA cells and the yield of rHSA A：Viable cell density. B：Viability. C：Glucose concentration. D：The productivity of rHSA was detected by ELISA（*P<0.05）

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