重组人尿激酶原冻干产品的稳定性  

Stability of Lyophilized Recombinant Human Prourokinase

作　　者：高丽华[1] 张正光[1] 胡显文[1] 胥照平[1] 肖成祖[1] 郗永义[1] 邵勇[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2008年第3期394-397,共4页Letters in Biotechnology

基　　金：国家高技术研究发展计划项目(2007AA021702;2006AA02A245)

摘　　要：目的:探讨重组人尿激酶原(rhPro-UK)冻干产品的稳定性。方法:采用S-2444发色底物法测定贮存在4℃和-20℃的rhPro-UK冻干产品的活性、单链比例随时间的变化规律;用SDS-PAGE及RP-HPLC肽图分析贮存在-20℃的rhPro-UK冻干产品的结构与组成的变化。结果:4℃保存3年后的rhPro-UK冻干产品的总活性和单链比例基本没有变化,但随着贮存时间的延长,有部分产品降解,如贮存78个月的样品,总活性可降低13%～15%;-20℃保存78个月后,rhPro-UK冻干产品的总活性和单链比例未见明显变化。SDS-PAGE及RP-HPLC肽图图谱显示,-20℃贮存78个月后的rhPro-UK冻干产品的组成和结构没有变化。结论:rhPro-UK冻干产品在4℃的贮存寿命可达3年;长期贮存于-20℃的rhPro-UK冻干产品,其总活性、单链比例及结构组成非常稳定。Objective： To study the stability of lyophilized recombinant human prourokinase（rhPro-UK）. Methods： The lyophilized rhPro-UK products stored at 4℃ and -20℃ were analyzed by S-2444 chromogenic assay to measure the total activity and the inactive single-chain zymogen form （scu-PA） ratio of the rhPro-UK. SDS-PAGE and RP-HPLC peptide mapping methods were used to determine the alterations of rhPro-UK composition and structure when stored at -20℃. Results： There was no influence in the activity and scu-PA ratio of lyophilized rhPro-UK stored at 4℃ during 3 years, but 13%-15% of its activity was lost when last for 78 months. Almost no alteration of the activity and scu-PA ratio was detected when the lyophilized products were kept at -20℃ for 78 months. At the same time, the results of SDS-PAGE and RP-HPLC peptide mapping showed that there was no change in composition and structure after the rhPro-UK stored at -20℃ for 78 months. Conclusion： The shelf life of lyophilized rhPro-UK products kept at 4℃ could reach 3 years, and the lyophilized rhPro-UK was very stable when stored at -20℃.

关 键 词：重组人尿激酶原 稳定性 冻干 贮存寿命 

分 类 号：Q502[生物学—生物化学] Q813