两种贝伐珠单抗生物学活性检测方法的对比

Comparison of two methods for biological activity assay of Bevacizumab

目的对比两种贝伐珠单抗生物学活性检测方法,比较两种方法的检测性能。方法对两种贝伐珠单抗生物学活性检测方法[人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖抑制法和荧光素酶报告基因法(reporter gene assay,RGA)]分别进行验证,比较两种方法的检测性能;采用两种方法同时检测53批贝伐珠单抗样品(包括原研药及其生物类似药),并对两种方法的稳定性指示能力进行分析。结果RGA法检测周期更短,曲线的R2、信噪比、线性段范围均优于HUVEC增殖抑制法,检测范围更宽,精密度更高;两种方法检测的贝伐珠单抗生物学活性差异无统计学意义(P>0.05);除强光照外,RGA法与HUVEC增殖抑制法相比,在反映贝伐珠单抗生物学活性变化方面相似或略好。结论RGA法的检测性能优于HUVEC增殖抑制法,且能更好地反映贝伐珠单抗生物学活性变化,可作为HUVEC增殖抑制法的替代方法,用于产品的放行检测及稳定性研究等。

Objective To compare the performances of two methods for biological activity assay of Bevacizumab.Methods Two methods for biological activity assay of Bevacizumab,human umbilical vein endothelial cell(HUVEC)proliferation inhibition assay and luciferase reporter gene assay(RGA),were validated to compare their detection performances.Fifty-three batches of Bevacizumab(including original patented drug and its biosimilar)were determined simultaneously by the two methods,and the stability indicating properties of the two methods was compared.Results RGA showed a shorter experiment period,wider detection range and higher precision,of which the R2 value of curve,signal/noise ratio and linear range were superior to those of HUVEC proliferation inhibition assay.No significant difference was observed between the biological activities of Bevacizumab determined by RGA and HUVEC proliferation inhibition assay(P>0.05).RGA was similar to or slightly better than HUVEC proliferation inhibition assay in reflecting the biological activity change of Bevacizumab except strong photostress.Conclusion The performance of RGA was superior to that of HUVEC proliferation inhibition assay.The method effectively reflected the biological activity change of Bevacizumab,thus may be used as an alternative to classic HUVEC proliferation inhibition assay for product release and stability test of the McAb.