抗EB病毒LMP2A单克隆抗体的制备及免疫学特性分析

Preparation and identification of immunologic characterization of a monoclonal antibody against latent membrane protein 2A of EBV

目的:利用杂交瘤技术制备抗EB病毒潜伏膜蛋白2A(LMP2A)单克隆抗体并分析其免疫学特性。方法:以LMP第2和第5胞外区表位串联制备的融合蛋白免疫BALB/c小鼠,取免疫后小鼠脾细胞与SP2/0细胞融合,制备抗LMP2A的单克隆抗体。通过ELISA、Western blot、免疫荧光、免疫组化、流式细胞术等方法鉴定单抗的免疫学特性。结果:获得1株稳定并持续分泌抗LMP2A单克隆抗体的稳定杂交瘤细胞株,所分泌的单克隆抗体5C12-C4-A6能够特异性识别鼻咽癌细胞膜表面的LMP2A蛋白。结论:本文制备了能够特异性识别鼻咽癌细胞膜表面LMP2A蛋白的单克隆抗体,为进一步进行鼻咽癌临床早期诊断和分子靶向治疗奠定了基础。

Objective：To produce a monoclonal antibody（mAb） against latent membrane protein 2A（LMP2A） with hybridoma technology and characterize its immune specificity. Methods：The spleen cells from BALB / c mice immunized with epitope fusion protein LMP2A were fused with SP2 / 0 cells to produce a monoclonal antibody against LMP2A. We further determined immunospecificity of the purified antibody by various immunological assays such as ELISA,Western blotting assays,immunofluorescence,immunohistochemisty and FACS. Results：We successfully produced a hybridoma cell line,which continuously and stably secreted anti-LMP2A mAb. Moreover,this mAb can effectively recognize the transmembrane glycoprotein LMP2A over-expressed in nasopharyngeal carcinoma cells and tissues. Conclusion：We successfully obtained mouse anti-LMP2A mAb that can specifically recognize the transmembrane glycoprotein LMP2A in nasopharyngeal carcinoma cells. It can be potentially applied to early stage clinical diagnosis and some tumor targeted therapy.