A reporter gene assays for bioactivity determination of human chorinonic gonadotropin
10.16438/j.0513-4870.2023-0545
- VernacularTitle:报告基因法测定人绒毛膜促性腺激素生物活性
- Author:
Ying HUANG
1
,
2
;
Xiao-ming ZHANG
3
;
He-yang LI
3
;
Lü-yin WANG
3
;
Hui ZHANG
3
;
Ping LÜ
3
;
Jing LI
3
;
Xiang-dong GAO
1
;
Cheng-gang LIANG
3
Author Information
1. China Pharmaceutical University, Nanjing 211198, China
2. National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China
3. National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China
- Publication Type:Research Article
- Keywords:
reporter gene assay;
human chorinonic gonadotropin;
four-parameter curve;
biological activity;
method validation
- From:
Acta Pharmaceutica Sinica
2024;59(2):432-438
- CountryChina
- Language:Chinese
-
Abstract:
This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.
