Establishment and validation of bioactivity measurement method for recombinant human midkine
10.16438/j.0513-4870.2023-0338
- VernacularTitle:重组人源中期因子生物活性测定方法的建立与验证
- Author:
Ying ZHANG
1
;
Ting HE
2
;
Ming-feng QIU
3
;
Sheng-bin PENG
2
Author Information
1. School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; Usynova Pharmaceuticals Ltd., Shanghai 201203, China
2. Usynova Pharmaceuticals Ltd., Shanghai 201203, China
3. School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China
- Publication Type:Research Article
- Keywords:
recombinant human midkine;
cck-8;
bioactivity;
proliferation;
method validation
- From:
Acta Pharmaceutica Sinica
2024;59(1):198-201
- CountryChina
- Language:Chinese
-
Abstract:
To establish and optimize a method for the detection of recombinant human midkine (rhMK) activity and verify its methodology, cell counting kit-8 (cck-8) method was used to measure the proliferation activity of rat knee chondrocytes. The specificity, accuracy, precision, linearity and robustness of the method were also verified in this study. The established method was proven to have good specificity because the buffer of rhMK and recombinant human interleukin-1 receptor antagonist have no obvious active effect; the recoveries of the samples with relative activities of 50%, 75%, 100%, 125%, 150% were in the range of 80.0% to 124.0% by statistical analysis, the relative standard deviations (RSD) of relative potency were all within 20%, the linear correlation coefficient, R2 ≥ 0.98, suggesting that the accuracy, precision and linearity of the method were good; the robustness correlation coefficient, R2 ≥ 0.92 and the ratio of maximum to minimum of sigmoidal dose-response were no less than 1.5, indicating that robustness of the methods was good. In conclusion, a bioactivity measurement method for rhMK was established and fully validated in this study and it provides a reliable method for the bioactivity analysis of rhMK routine samples during the development. This study was approved by the Animal Care and Use Committee of Shanghai Model Organisms Center, Inc. (approval number: 2019-0008-06).
