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目的分析重组人粒细胞刺激因子生物学活性检测中,以NFS-60细胞为测活基础的几种常见显色/裂解/检测方法,所得检测结果是否一致。方法应用NFS-60细胞/MTT染色法、NFS-60细胞/MTS染色法、NFS-60细胞/CCK-8染色法、NFS-60细胞/化学发光法等检测重组人粒细胞刺激因子生物学活性,并对结果进行统计分析。统计分析了NFS-60细胞/MTT法双波长(570 nm检测,630 nm参比)、单波长(570 nm检测和630 nm检测)结果。分析比较了NFS-60细胞/MTS动态检测法、NFS-60细胞/CCK-8动态法检测结果。结果各生物学活性检测方法的检测结果差别不显著(P>0.05);NFS-60/MTT法双波长、单波长检测结果差别不显著(P>0.05)。NFS-60细胞/MTS动态检测法、NFS-60细胞/CCK-8动态法与NFS-60细胞/MTT染色法检测结果间差别不显著(P>0.05)。结论以NFS-60细胞为测活基础的几种常见的显色/裂解/检测方法,所得检测结果一致,为各实验室(应用不同显色/裂解/检测方法)给出的数据是否可以综合利用提供依据,为2015年版《中国药典》中重组人粒细胞刺激因子生物学活性检测方法的扩充提供支持,也为其他细胞因子生物学活性检测方法的扩充提供借鉴。
OBJECTIVE: To analyze the common chromogenic / lytic / detection methods based on the assay of NFS-60 cells in the detection of biological activity of recombinant human granulocyte stimulating factor. The results obtained are consistent. Methods The biological activity of recombinant human granulocyte-stimulating factor was detected by the method of NFS-60 / MTT staining, NFS-60 / MTS staining, NFS-60 / CCK-8 staining and NFS-60 / chemiluminescence , And the results of statistical analysis. The results of double wavelength (570 nm detection, 630 nm reference) and single wavelength (570 nm detection and 630 nm detection) by NFS-60 cell / MTT method were statistically analyzed. The results of NFS-60 cell / MTS dynamic assay and NFS-60 cell / CCK-8 assay were compared and analyzed. Results There was no significant difference between the two methods (P> 0.05). The results of double wavelength and single wavelength of NFS-60 / MTT method showed no significant difference (P> 0.05). There was no significant difference between NFS-60 / MTS assay, NFS-60 / CCK-8 assay and NFS-60 / MTT assay. Conclusions Several common chromogenic / lytic / detection methods based on the measurement of NFS-60 cells are consistent with the results obtained for each laboratory (using different chromogenic / lytic / detection methods) whether the data can be synthesized It provides support for the expansion of the detection methods of the biological activity of recombinant human granulocyte stimulating factor in the Chinese version of the Chinese Pharmacopoeia 2015 edition and provides references for the expansion of other cytokine bioassay methods.