重组人胸腺素β4生物学活性测定方法的建立  

Development of a Biological Activity Detection Method for Recombinant Human Thymosin β4

作　　者：徐宏伟[1] 张俊英 马杉姗[1] 汤晓闯[1] 马素永[1,2] 聂李亚 许松山[1,2] XU Hong-Wei;ZHANG Jun-Ying;MA Shan-Shan;TANG Xiao-Chuang;MA Su-Yong;NIE Li-Ya;XU Song-Shan(Beijing Northland Biotech. Co., Ltd., Beijing 100085,China;Beijing Naked Plasmid Gene Therapy Drug Engineering Technology Research Center, Beijing 100085,China)

机构地区：[1]北京诺思兰德生物技术股份有限公司,北京100085 [2]北京市裸质粒基因治疗药物工程技术研究中心,北京100085

出　　处：《生物技术通讯》2017年第5期661-664,共4页Letters in Biotechnology

摘　　要：目的:建立适用于重组人胸腺素β4(rhTβ4)体外质量控制的生物学活性测定方法,用于该制品的生物学活性评价。方法:利用ECV304细胞迁移法,摸索细胞密度、玻连蛋白(Vn)浓度、rhTβ4浓度等影响因素,并对试验条件进行优化及方法验证。结果:细胞密度为0.5×104/孔、Vn浓度为12.5μg/mL、rhTβ4浓度为200nmol/L、孵育时间24h和细胞迁移时间4h时,细胞迁移率较高。结论:建立了rhTβ4生物学活性测定方法,该方法专属性强、重复性好、准确性高,可用于rhTβ4生物学活性测定。Objective:To control the quality of recombinant human thymosin beta4(rhTβ4)in vitro,the biological activity detection method was developed and evaluated.Methods:The ECV304cell migration method wasused to detect the biological activity of rhTβ4.The key factors including cell density,vitronectin(Vn)concentration,rhTβ4concentration were screened to establish the optimal migration conditions.Results:The highest cell migration rate was observed under the conditions of the cell density0.5×104/well,Vn concentration12.5μg/mL,rhTβ4concentration200nmol/L,incubation time24h and cell migration time4h.Conclusion:The developed biological activity detection method of rhTβ4has high specificity,accuracy and repeatability,and it can be used for thequality control of rhTβ4.

关 键 词：重组人胸腺素β4 活性检测 细胞迁移 

分 类 号：Q502[生物学—生物化学] Q24