Study on equivalence of biological activity of insulin glargine by quantitative immunofluorescence assay and insulin bioassay
- VernacularTitle:甘精胰岛素生物学活性定量免疫荧光测定法和胰岛素生物测定法的等效性研究
- Author:
Yi-Min GAO
1
,
2
;
Hong-Mei ZHANG
;
Kai-Yong HE
;
Deng-Ke YIN
;
Bei SUN
;
Lei-Ming XU
Author Information
- Keywords: biological activity; insulin glargine; quantitative immunofluorescence assay; transgenic cells; methodology vali-dation; equivalence evaluation
- From: Chinese Pharmacological Bulletin 2024;40(11):2193-2199
- CountryChina
- Language:Chinese
- Abstract: Aim To establish a quantitative immunofluorescent bioactivity assay(ICW)for insulin glargine based on CHO-IN-SRB 1284 transgenic cells,and to study its equivalence with in-sulin bioassay of Ch.P.Methods The cells were diluted 25 times with 1.5 × 108 L-1 cell density plates and 1 500 μmol·L-1 insulin glargine,and then diluted with a 3-fold gradient se-ries.The cells were stimulated in microporous plates for 20 min.After fixation,permeation and antibody incubation.Quantitative immunofluorescence biological activity was detected by odyssey two-color infrared fluorescence imaging system.Results There was a good dose-effect relationship between the concentration of insulin glargine in ICW and its relative potency.The method had good specificity,and the relative accuracy,intermediate preci-sion and linearity met the requirements.The relative deviation of biological activity results of 7 batches of insulin glargine samples measured by the two methods was less than 10％.The results were analyzed by SPSS and SAS software,which showed that the methods were correlated and equivalent.Conclusions The quantitative immunofluorescence assay for the biological activity of insulin glargine can be established.The method has good spe-cificity,high accuracy and precision,and has correlation and e-quivalent with biotiter assay,which can be applied to in vitro ef-ficacy evaluation and quality control of insulin glargine.
