Optimization of expression conditions of modified tissue plasminogen activator
10.13200/j.cnki.cjb.004624
- VernacularTitle:组织型纤溶酶原激活剂改构体表达条件的优化
- Author:
Ziwei YANG
1
Author Information
1. State Key Laboratory of Druggability Evaluation and Systematic Translational Medicine, Tianjin Institute of Pharmaceutical Research, Tianjin 300301, China
- Publication Type:Journal Article
- Keywords:
Tissue plasminogen activator(t-PA);
Modified t-PA(mt-PA);
Sodium butyrate;
Culture temperature;
CHOZN cell line
- From:
Chinese Journal of Biologicals
2025;38(12):1483-1489+1497
- CountryChina
- Language:Chinese
-
Abstract:
Objective To optimize the expression conditions of modified tissue plasminogen activator(t-PA) in order to enhance the yield and quality of modified t-PA(mt-PA).Methods The fed-batch cultivation method was used to detect the effects of low temperature culture(37 ℃ for 5 days and cooled to 30 ℃) and different concentrations of sodium butyrate(0,0. 5, 1, 2, 5 mmol/L) on the growth status of stable Chinese hamster ovarian cancer mt-PA cell line(CHOZN-mt-PA), the concentration of key metabolites(glucose, lactic acid, and ammonia) of the cells, the expression level of mt-PA, and the ratio of single/double chains. The thrombolytic activity of mt-PA in vitro was measured by bubble method.Results The cell proliferation was inhibited, the glucose consumption as well as the accumulation of lactic acid and ammonia were reduced, the expression level of mt-PA greatly increased, and the ratio of single/double chains was stable with the addition of 2 mmol/L sodium butyrate and 30 ℃ low-temperature cultivation. Compared with the corresponding control group, there was no statistically significant difference in thrombolytic activity of mt-PA prepared at 30 ℃ with the addition of 2 mmol/L sodium butyrate in vitro(t = 0. 546 32 and 0. 441 70, P = 0. 613 9 and 0. 681 5, respectively).Conclusion Cultivation at 30 ℃ and the addition of 2 mmol/L sodium butyrate can increase the expression level of mt-PA without reducing its thrombolytic activity in vitro,which provides a reliable strategy for the development of high-yield culture process of mt-PA.
