抗Nodal单克隆抗体的制备及夹心ELISA试剂盒的研制

Preparation of Nodal Antibody and Development of Its ELISA Kit

利用Nodal成熟肽为抗原免疫小鼠,取其脾脏B淋巴细胞与骨髓瘤细胞SP2/0融合,经间接ELISA法、融合细胞有限稀释法克隆筛选出阳性单克隆杂交瘤细胞株,接种至小鼠腹腔,培养后抽取腹水并用饱和硫酸铵沉淀法初步纯化,从而获得抗Nodal单克隆抗体。该单克隆抗体经特异性验证实验后,与Nodal的多克隆抗体兔血清一道分别作为夹心ELISA检测试剂盒的上下层抗体使用,建立了以Nodal为抗原的夹心ELISA试剂盒的检测方法,并进行了对肿瘤细胞提取物、肿瘤小鼠血清的检测。该研究为该试剂盒在肿瘤诊断方面的应用奠定了基础。

Nodal mature peptide was used as antigen to immunize the mouse. By cell fusion between B lymphocyte and SP2/0, indirect ELISA and limiting dilution assay, the positive monoclonal hybridoma cell was obtained. Then the hybridoma cell was inoculated into enterocoelia of mouse and extracted after amplification, which was purified by sedimentation of saturation ammonium sulfate. The monoclonal antibody, after test for the specificity, was used with another Nodal polyclonal antibody of rabbit serum in the sandwich ELISA kit. The test method of the kit was established then and its pre- liminary application had been researched by examining many sorts of tumor extractive as well as the serum of mice with tumor.