| 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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| 引用本文： | 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,2014(7):875-882. |
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| 作者姓名： | 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 |
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| 作者单位： | 江苏大学基础医学与医学技术学院,江苏 镇江 212013;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;南京医科大学第二附属医院病理科,江苏 南京 210011;江苏大学基础医学与医学技术学院,江苏 镇江 212013;南京医科大学第二附属医院病理科,江苏 南京 210011;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;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 210030;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 210031;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;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 138670;江苏大学基础医学与医学技术学院,江苏 镇江 212013;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;南京医科大学第二附属医院病理科,江苏 南京 210011 |
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| 基金项目： | 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) |
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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?
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| 关 键 词： | EB病毒 LMP2A 单克隆抗体 免疫学特性 |
| 收稿时间： | 3/6/2014 12:00:00 AM |
Preparation and identification of immunologic characterization of a monoclonal antibody against latent membrane protein 2A of EBV |
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| Xu Xin,Tang Xiaojun,Xiong Lin,Wang Huan,Li Wenjie,Gao Chang,Xiong Siping,Qiu Zhenning,Tan Min Han,Vo Jess Honganh,Feng Zhenqing and Chen Renjie.Preparation and identification of immunologic characterization of a monoclonal antibody against latent membrane protein 2A of EBV[J].Acta Universitatis Medicinalis Nanjing,2014(7):875-882. |
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| Authors: | Xu Xin Tang Xiaojun Xiong Lin Wang Huan Li Wenjie Gao Chang Xiong Siping Qiu Zhenning Tan Min Han Vo Jess Honganh Feng Zhenqing and Chen Renjie |
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| Institution: | School of Medical Science and Laboratory Medicine,Jiangsu University,Zhenjiang 212013;Department of Pathology,Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Department of Pathology,the Second Affiliated Hospital of NJMU,Nanjing 210011;School of Medical Science and Laboratory Medicine,Jiangsu University,Zhenjiang 212013;Department of Pathology,the Second Affiliated Hospital of NJMU,Nanjing 210011;Department of Pathology,Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Department of Pathology,Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210030;Department of Pathology,Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210031;Institute of Bioengineering and Nanotechnology,Division of Biodevices and Diagnostics,Singapore 138669;Institute of Bioengineering and Nanotechnology,Division of Biodevices and Diagnostics,Singapore 138669;School of Medical Science and Laboratory Medicine,Jiangsu University,Zhenjiang 212013;Department of Pathology,Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Department of Pathology,the Second Affiliated Hospital of NJMU,Nanjing 210011 |
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| 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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| Keywords: | EBV LMP2A monoclonal antibody immune specificity |
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