细胞增殖法检测注射用特立帕肽生物活性方法的建立和验证

Establishment and validation of the method for determination of biological activity of teriparatide for injection by cell proliferation test

目的:建立并验证检测注射用特立帕肽生物活性的细胞增殖法。方法:采用MC3T3-E1细胞增殖法检测不同浓度注射用特立帕肽的生物活性,通过检测特异性拮抗剂TIP-39对特立帕肽促MC3T3-E1细胞增殖的抑制作用以验证方法的专属性,同时验证方法的重复性、线性和检测范围并用确立的方法将不同批次的注射用特立帕肽与上市对照品进行活性对比。结果:建立的细胞增殖法专属性和重复性良好,有效检测范围是1.25～80 nmol.L-1,促增殖活性为20%～85%,线性相关系数=0.989 8,且自制注射用特立帕肽活性与上市对照品一致。结论:建立的细胞增殖法能有效用于注射用特立帕肽的体外生物活性测定。

OBJECTIVE To establish and validate the cell proliferation test for detecting the biological activity of teriparatide for injection.METHODS MC3T3-E1 cell proliferation test was used for detecting biological activity of teriparatide for injection in different concentration.The specificity investigation of teriparatide stimulating MC3T3-E1 cell proliferation was demonstrated by adding the specific inhibitor TIP-39.Reproducibility,linear and range of this method were validated and the activity consistency evaluation of different teriparatide batches with launched product was implemented.RESULTS The cell proliferation test had good specificity and reproducibility and the detection range was from 1.25 to 80 nmol·L-1,the biological activity range of teriparatide was from 20% to 85%,the linear correlation coefficient was 0.9898.Furthermore,the biological activity of domestic produced teriparatide for injection was consistent with launched product.CONCLUSION The established method can be applied to the activity determination of teriparatide for injection in vitro effectively.