重组人粒细胞集落刺激因子生物学活性测定方法研究

Study on determination method of the rhG-CSF biological activity

目的用世界卫生组织国际标准品粒细胞集落刺激因子 (G CSF)生物学活性标准品研究NFS 6 0细胞测定重组人粒细胞集落刺激因子 (rhG CSF)生物学活性的方法。方法采用溴化四唑蓝 (MTT)染色法。结果NFS 6 0细胞染色体为 39条 ;细胞数在 1× 10 5～ 1× 10 7个 /ml间MTT染色显示梯度 ;将 (4 ,4)法引入rhG CSF生物学活性的测定 ,单次实验结果的平均可信限率为 13.5 6 0 % ,(4 ,4)法测定rhG CSF生物学活性的批内CV <10 % ,批间CV为 10 .10 9%。结论 (4 ,4)法可用于rhG CSF生物学活性的测定 ,对测定方法标准化后 ,测定结果可准确、有效地指导rhG CSF的生产和研究。

PurposeThe determination method of rhG CSF biological activity by NFS 60 cells was studied in accordance with WHO international standard for G CSF.MethodsMTT method was adopted.ResultsThe chomosone number of NFS 60 cell was 39, about 1×10 5 cells/ml to 1×10 7 cells/ml,the dying color showed gradient when the NFS 60 cells was dyed by MTT,the method(4,4) was adopted in the determination of rhG CSF biological activity,the average FL% of potency was 13.560% for single expriment,and the CV of inter assay and intra assay was lower than 10% and 10.109%,respectively.Conclusionmethod(4,4) can be used in the determination of rhG CSF biological activity,and the results can guide the study and the manufacture of rhG CSF effectively.