基于抑制单核巨噬细胞增殖活性检测的注射用骨肽质量控制

目的:通过建立注射用骨肽抑制单核巨噬细胞(THP-1)增殖的生物活性测定法,以补充完善其质量控制方法。方法:采用CCK-8法以体外抑制THP-1增殖作用来考察注射用骨肽的生物活性。以A01160410批次注射用骨肽作为对照物质进行效价测定。结果:注射用骨肽对THP-1的增殖具有一定的抑制作用。采用量反应平行线(2·2)法作为注射用骨肽抑制THP-1增殖的生物效价检测方法,重复性较好,样品检测结果均能通过可靠性检验(回归P0.01,偏离平行P0.05)。A20160102批次注射用骨肽效价94.50 U·m L-1,可信限率(FL)=6.14%,可信限范围为88.87~100.47 U·m L-1;A01160401批次注射用骨肽效价为92.04 U·m L-1,FL=4.36%,可信限范围为88.12~96.15 U·m L-1;A01160406批次注射用骨肽效价为83.25 U·m L-1,FL=8.52%,可信限范围为76.46~90.65 U·m L-1。3个批次注射用骨肽实验结果合并分析结果显示,注射用骨肽抑制THP-1的效价为90.37 U·m L-1,FL=8.63%,可信限范围为82.91~98.51 U·m L-1。不同操作人员对注射用骨肽的效价测定结果没有明显差别,具有良好的可重复性。结论:该研究建立的生物效价检测方法可以补充作为注射用骨肽生物评价的质控方法之一。

Quality Control of Ossotide for Injection in Inhibiting Proliferation of Monocyte-macrophages

Objective: To supplement and improve the quality control method hrough a bioassay test about ossotide for injection on inhibiting the proliferation of monocyte-macrophages(THP-1). Method: The biological activity of ossotide for injection was investigated by its inhibitory effect on THP-1 proliferation in vitro with CCK-8 assay. A01160410 batch of ossotide for injection was used as standard control substanceunder the potency determination. Result: The ossotide for injection had a significant inhibitory effect on the proliferation of THP-1.The parallel line(2·2)assay method as the biopotency detection method for inhibitory effect of ossotide for injection on the proliferation of THP-1,had a good repeatability. All the samples passed the reliable test (regression P<0.01,deviation from parallel P>0.05). A20160102 batch of ossotide for injection potency was 94.50 U·mL^-1,with confidence limit rate(FL)=6.14%,and the confidence limit range was from 88.87 U·mL^-1 to 100.47 U·mL^-1;A01160401batch of ossotide for injection potency was 92.04 U·mL^-1 with FL=4.36%,and the confidence limit ranged from 88.12 to 96.15 U·mL^-1. A01160406 batch of ossotide for injection potency was 83.25 U·mL^-1 with FL=8.52%, and the confidence limit ranged from 76.46 to 90.65 U·mL^-1. The combined analysis of the experimental results of three batches of ossotide for injection showed that the potency of ossotide for injection for inhibiting THP-1 was 90.37 U·mL^-1 with FL=8.63%,and the confidence limit ranged from 82.91 to 98.51 U·mL^-1. Different operators had no significant difference in the potency result of ossotide for injection,with a good repeatability. Conclusion: The biopotency detection method established in our study can be used as one of the quality control methods for the bioassay of ossotide for injection.