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

作　　者：徐鑫[1] 唐小军[2] 熊林[3] 王欢[1] 李文杰[3] 高畅[2] 熊四平[2] 仇镇宁[2] Tan Min Han Vo Jess Honganh 冯振卿[1,2] 陈仁杰[3] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013 [2]南京医科大学病理学系,卫生部抗体技术重点实验室,江苏南京210029 [3]南京医科大学第二附属医院病理科,江苏南京210011 [4]新加坡生物工程和纳米技术研究院生物医疗器械和诊断中心,新加坡138669

出　　处：《南京医科大学学报（自然科学版）》2014年第7期875-882,共8页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省临床医学科技专项(BL2013038);江苏省普通高校研究生科研创新基金(CXZZ12_0563)

摘　　要：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.

关 键 词：EB病毒 LMP2A 单克隆抗体 免疫学特性 

分 类 号：R392.11[医药卫生—免疫学]