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以重组人甘露聚糖结合凝集素(MBL)相关丝氨酸蛋白酶2-C端(MASP2-C)蛋白为抗原,免疫Balb/C小鼠,制备功能性单克隆抗体。获得4株能稳定分泌抗人MASP2-C单克隆抗体的细胞株;其亚型均属于IgG1,轻链为κ链;ELISA、western blot鉴定其特异性良好;ELISA显示其与天然MASP2结合活性好;C4b沉积实验证实其对补体凝集素途径具有阻断作用。因此,我们成功制备了具有较好功能活性的抗MASP2-C蛋白单克隆抗体,为研究MASP2的生物学功能及MASP2缺损有关疾病的诊断提供了有价值的工具和手段。
Functional monoclonal antibodies were prepared by immunizing Balb / C mice with the recombinant human mannan-binding lectin (MBL) -related serine protease 2-C (MASP2-C) protein as antigen. Four stable cell lines secreting anti-human MASP2-C monoclonal antibody were obtained. The subtypes were all IgG1, the light chain was κ chain, and the specificity was identified by ELISA and western blot. ELISA showed that the binding activity with MASP2 was good ; C4b deposition experiments confirmed that it has a blocking effect on the complement lectin pathway. Therefore, we successfully prepared the monoclonal antibody against MASP2-C protein with good functional activity, which provided valuable tools and methods for the study on the biological function of MASP2 and the diagnosis of MASP2-related diseases.