Comparison of several common methods for measuring the biological activity of recombinant human granulocyte colony stimulating factor
- Author:
Xin-Chang SHI
1
Author Information
- Publication Type:Journal Article
- Keywords: Biological activity; CCK-8; Dual wavelength detection; MTS; MTT; NFS-60 cells; Recombinant human granulocyte colony stimulating factor; Single wavelength detection
- From: Chinese Pharmaceutical Journal 2016;51(13):1080-1084
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To compare several common staining and detection methods using NFS-60 cells for biological activity test of recombinant human granulocyte colony stimulating factor (rhG-CSF). METHODS: The biological activity of rhG-CSF was detected using some common methods, named NFS-60 cells/MTT staining, NFS-60 cells/MTS staining, NFS-60 cells/CCK-8 staining, and NFS-60 cells/fluorescence staining. The biological activity was detected using the NFS-60 cells/MTT method using dual wavelength (570 nm detection, 630 nm reference) and single wavelength (570nm detection and 630nm detection). The biological activity was detected using NFS-60 cells/MTS dynamic detection method and NFS-60 cell/CCK-8 dynamic method. Then, the results were analyzed and compared. RESULTS: The different methods were not significantly different (P>0.05); the difference between the dual wavelength detection and single wavelength detection of NFS-60 cells/MTT was not significant (P>0.05). The results from NFS-60 cells/MTS dynamic detection method, NFS-60 cells/CCK-8 dynamic method and NFS-60 cells/MTT method had not significant difference (P>0.05). CONCLUSION: The biological activity determination results of the tested methods using NFS-60 cells are consistent. This study provides basis for utilization of results from different laboratories using different methods, support for expansion of the biological activity detection method of rhG-CSF in Chinese Pharmacopoeia, and reference for expansion of the biological activity detection method of other cytokines.
