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利用G-CSF依赖型细胞株N8F-60,尝试将MTT比色法应用于基因工程产品重组人粒系集落刺激因子(rhG-CSF)的活性检测。多次实验探索出最适的细胞浓度和培养时间,细胞数为低于1×10 ̄4/孔,培养时间为30─48h。在宽的线性范围内检测G-CSF样品的生物活性,结果经传统的CFU-G方法验证可靠,且较后者更灵敏、简便、省时、省力,并可定性、定量地检测样品活性。特别适用于大量样品的同时检测。另外,此方法可避免接触放射性。
Using the G-CSF-dependent cell line N8F-60, MTT assay was applied to detect the activity of recombinant human granulocyte colony-stimulating factor (rhG-CSF). Multiple experiments to explore the most appropriate cell concentration and culture time, the number of cells is less than 1 × 10 ~ 4 / hole, culture time is 30 ─ 48h. The bioactivity of G-CSF samples was tested over a wide linear range. The results were validated by the conventional CFU-G method and more sensitive, simple, time-saving, labor-saving and qualitatively and quantitatively tested. Particularly suitable for the simultaneous detection of a large number of samples. In addition, this method avoids contact with radioactivity.