抗EB病毒LMP2A单克隆抗体的制备及免疫学特性分析
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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)


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

    Abstract:

    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 immuno-specificity 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.

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徐 鑫,唐小军,熊 林,王 欢,李文杰,高 畅,熊四平,仇镇宁,Tan Min Han, Vo Jess Honganh,冯振卿,陈仁杰.抗EB病毒LMP2A单克隆抗体的制备及免疫学特性分析[J].南京医科大学学报(自然科学版),2014,(7):875-882

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  • 收稿日期:2014-03-06
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  • 在线发布日期: 2014-07-05
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