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

VernacularTitle:聚乙二醇化Exendin-4类似物受体亲和力测定方法的建立
Author: Linfeng GUO ¹ ; Xiaojing HU ; Lei YIN ; Hong TIAN ; Wenbing YAO
Author Information

1. 中国药科大学生命科学与技术学院
Publication Type:Journal Article
Keywords: Exendin-4; PEGylation; determination of receptor binding affinity; luciferase reporter gene method
From: Journal of China Pharmaceutical University 2016;47(2):222-227
CountryChina
Language:Chinese
Abstract: 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. CHO-GLP-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.