重组人纽兰格林生物学活性检测方法及质控标准

目的　研究重组人纽兰格林生物学活性检测方法 ,并制定其质量控制标准。方法　采用激酶受体活化的酶联免疫吸附试验 (KIRA ELISA) ,通过优化细胞浓度、样品稀释梯度、药物刺激时间及细胞裂解方法等建立定量测定重组人纽兰格林体外生物学活性的方法 ,同时建立了一套完整可行的质量控制标准。结果　重组人纽兰格林刺激MCF 7细胞表面ErbB2 ErbB3分子磷酸化的反应存在量 效关系 ,相关系数大于 0 98,原液的平均比活性为 85 10U mg± 115 0U mg。并对原液设立肽图、纯度等检测项目 ,对成品设立热原质试验、鉴别试验等检测项目。结论　KIRA ELISA可用于定量检测重组人纽兰格林的体外生物学活性 ,结果准确可靠 ,重复性好 ;建立的方法和标准可以有效控制重组人纽兰格林制品的质量。

Method for Determination of Biological Activity and Standard for Quality Control of Recombinant Human Neuregulin

Objective To study the method for determination o f biological activity and the standard for quality control of recombinant human neuregulin.Methods Develop a kinase receptor activation enzyme -linked immunosorbant assay (KIRA-ELISA) by optimizing the concentration of MC F-7 cells,dilution gradient of sample,time for stimulating the phosphorylation of cells and the method for lysis of cells.Meanwhile,a practical standard for qu ality control was developed on the basis of physical examination and chemical te st on recombinant human neuregulin. Results Recombinant human neuregulin showed a dose-dependent stimu lating effect on the phosphorylation of ErbB3 molecules on the surface of MCF-7 cells (r>0.98).The mean specific activity of bulk was 8 510 U/mg±1 150 U/mg. A standard for quality control of recombinant human neuregulin,including the pep tide mapping and purity test on bulk and sterility and identity tests on final p roduct,was developed. Conclusion The developed KIRA-ELISA can be used for the quantitative d etermination of in vitro biological activity of recombinant human neuregulin.The result was accurate and reliable and of good reproducibility.The developed stan dard was effective in the quality control of recombinant human neuregulin.