聚乙二醇化Exendin-4类似物受体亲和力测定方法的建立  

Establishment and verification of assay method for the receptor binding affinity of a PEGylated Exendin-4 analogue

作　　者：郭林峰[1] 胡晓静[1] 印蕾[1] 田浤[1] 姚文兵[1] 

机构地区：[1]中国药科大学生命科学与技术学院,南京210009

出　　处：《中国药科大学学报》2016年第2期222-227,共6页Journal of China Pharmaceutical University

基　　金：国家"重大新药创制"科技重大专项资助项目(No.2012ZX09103101-016);教育部博士点基金资助项目(No.20120096110007)~~

摘　　要：基于荧光素酶报告基因法和本实验室前期构建的CHO-GLP-1R-CRE-Luc＋检测模型细胞株,建立并优化了长效降糖多肽——聚乙二醇化Exendin-4类似物PE与GLP-1受体的亲和力测定方法,并对其进行了方法学验证。结果表明,当药物作用4 h,化学发光底物作用15 min,检测药物在5.7×10-3-1.5×103nmol/L浓度区间内时,该检测方法的专属性和耐用性良好、准确度和精密度较高,符合《生物制品质量控制分析方法验证技术一般原则》的相关要求。本研究为该类GLP-1受体激动剂药物的快速评价与筛选奠定了基础。The assay method of GLP-1 receptor binding affinity for a long-acting hypoglycemic peptide—PEgylated Exendin-4 analogue（ PE） was optimized and established based on the luciferase reporter gene approach. CHOGLP-1R-CRE-Luc＋cells were previously constructed in our lab followed by the verification of methodology. This assay method showed good specificity and robustness as well as high accuracy and precision when PE was incubated with the cell for 4 h,the luminescent substrate reacted with cell lysates for 15 min and the concentration for PE ranged 5. 7 × 10- 3-1. 5 × 103 nmol / L,on which condition this developed method is in accordance with General Principles of Analytical Method Validation Techniques for Biological Products Quality Control. This study also lays the foundations for rapid evaluation and screening of GLP-1 receptor agonist drugs.

关 键 词：EXENDIN-4 聚乙二醇化 受体亲和力测定 荧光素酶报告基因法 

分 类 号：R392-33[医药卫生—免疫学]