抗人脑胶质瘤单克隆抗体的制备及识别抗原的提取

目的：制备新的抗人脑胶质瘤单克隆抗体（monoclonal antibody,McAb)，并提取其识别抗原。方法：以人脑恶性胶质瘤细胞系SHG－44为抗原免疫Balb/c小鼠，通过杂交瘤技术获得稳定分泌抗胶质瘤McAb的杂交瘤细胞株。以酶联免疫吸附测定及免疫组化等方法研究McAb的特性。通过亲和层析法提取该McAb识别的抗原。结果：得到1株稳定分泌抗体、效价高的杂交瘤细胞株，命名为SU－2000。鉴定表明该McAb属IgGl亚类，效价高，特异性高。识别的抗原位于胞膜，可能属于神经外胚层抗原；通过亲和层析法成功地提取出该抗原，十二脂硫酸3钠－聚丙烯酰胺凝胶电泳证实提取的抗原纯度高，分子量约为75kDa。结论：SU－2000 McAb及其识别抗原可进一步用于相关实验研究。

Establishment of the monoclonal antibody against human brain glioma and extraction of the recognized antigen

Objective To establish the new monoclonal antibody (McAb) against human brain glioma and to extract the antigen recognized by the McAb. Methods The Balb/c mice were immunized with the malignant human brain glioma cell SHG-44 as antigen and the hybridoma cell line secreting anti-glioma McAb steadily was obtained by using hybridoma technique. The property of the McAb was studied with enzyme-linked-immunosorbent assay and immunohistochemistry methods. The antigen recognized by the McAb was extracted by using affinity chromatograph. Results A hybridoma cell line secreting McAb steadily with high titer was obtained, which was named SU-2000. The McAb with high titer and specificity was proved to belong to the IgG1 subtype. The antigen recognized by the McAb was located on the cell membrane and might belong to the neuroectodermal antigen. The antigen was extracted successfully by using affinity chromatograph, and proved to have high quality with 75 kDa molecular weight by sodium dedecyl sulfate-polyacrylamide gel electrophoresis. Conclusion The SU-2000 McAb and the recognized antigen are useful for the further research.