基于Nb2-11细胞增殖的重组人生长激素生物学活性测定方法考察

Investigation on Biological Activity Assay of Human Recombinant Growth Hormone Based on Nb2-11 Cell Proliferation

该研究建立了基于细胞的重组人生长激素(rhGH)生物学活性检测方法,以替代动物体内试验法。首先,用不同浓度的rhGH刺激大鼠淋巴瘤细胞株(Nb2-11细胞)增殖,采用发光活细胞检测系统测定细胞增殖情况,并利用四参数拟合分析计算样品相对效价。随后,对试验条件进行了优化,并对方法的专属性、相对准确度、精密度和线性进行验证,结果显示方法专属性较好,相对准确度的相对偏倚范围为–6.2%~0.9%,线性回归方程相关系数为0.9850,中间精密度几何变异系数范围为5.2%~7.7%,线性范围为50%~200%。最后用该方法和动物体内试验法分别检测15批rhGH原液的生物学活性,结果显示该方法检测结果与动物体内试验法检测结果的一致性良好。该研究建立并验证了测定rhGH生物学活性的Nb2-11细胞增殖法,可作为rhGH生物学活性的常规检测方法用于rhGH的生物学活性评价和质量控制。

A cell-based method for recombinant human growth hormone(rhGH)bioactivity evaluation was established to replace in vivo animal experiments.Different concentrations of rhGH were used to stimulate the proliferation of the Nb-2 rat lymphoma cell line(Nb2-11 cell)and luminescent cell viability assay was used to measure the cell proliferation to evaluate the bioactivity of rhGH.The relative potency of rhGH was calculated by the 4-parameter logistic(4PL)regression model.Then,the method parameters were optimized.The specificity,relative accuracy,precision,and linearity of this method were verified.The results showed that this method had good specificity,and the relative bias for relative accuracy range was–6.2%-0.9%,the correlation coefficient of the linear regression equation was 0.9850,the geometric coefficient of variation for intermediate precision range was 5.2%-7.7%,the linear range was 50%-200%.The biological activities of 15 batches of rhGH products were investigated by the Nb2-11 cell proliferation method and the in vivo animal experiment,respectively.The results showed that these two methods had a good consistency.In this study,the Nb2-11 cell proliferation method for determination of rhGH bioactivity was established and verified,and it could be used as a routine method for rhGH bioactivity evaluation and quality control.