Determination of biological activity of human insulin by a homogeneous time-resolved fluorescence method
10.16438/j.0513-4870.2022-0641
- VernacularTitle:均相时间分辨荧光法测定人胰岛素生物学活性
- Author:
Lü-yin WANG
1
;
Yan-feng YANG
2
;
Xiao-ming ZHANG
1
;
Ping LÜ
1
;
Hui ZHANG
1
;
Jing LI
1
;
Cheng-gang LIANG
1
Author Information
1. NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Division of Hormone, National Institutes for Food and Drug Control, Beijing 102629, China
2. NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Division of Hormone, National Institutes for Food and Drug Control, Beijing 102629, China; China Pharmaceutical University, Nanjing 211198, China
- Publication Type:Research Article
- Keywords:
biological activity;
human insulin;
transgenic cell;
homogeneous time resolved fluorescence;
method validation
- From:
Acta Pharmaceutica Sinica
2022;57(10):3223-3228
- CountryChina
- Language:Chinese
-
Abstract:
We developed an in-vitro bioassay for determining the bioactivity of human insulin by homogeneous time-resolved fluorescence immunoassay. CHO-INSR B1284 transgenic cells were used as target cells. Key assay parameters, including the cell density, the range of working concentrations, and the stimulation time were optimized. The specificity, relative accuracy, intermediate precision, linearity, and range of the method were validated, as well as the passage stability of the CHO-INSR B1284 cell line. The national standard of recombinant human insulin was used as the benchmark to evaluate the relative potency of insulin analogues and drugs. The drugs and the reference human insulin showed a dose-response relationship, R2 > 0.995, which conforms to the four-parameter equation: y = (A - D) / [1 + (x/C)B] + D. Specificity of the method was good. The geometric mean, relative bias, and geometric coefficient of variation (GCV, %) of the five concentrations (n = 8, 64%, 80%, 100%, 125% and 156%) met the requirements of the General Rules of Chinese Pharmacopoeia, 2020 edition, Volume IV (9401). In summary, a bioassay for determining the in vitro bioactivity of human insulin based on a homogeneous time-resolved fluorescence technique was established; the method was simple, time-saving, accurate and precise, and could be used for the evaluation of biological activity and quality.
