| 抗Ⅳ型登革病毒NS1单克隆抗体的制备及特性鉴定 |
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| 作者姓名： | Ding XX DI B Che XY |
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| 作者单位： | 1. 南方医科大学珠江医院医学检验中心,广东,广州,510282
2. 广州市疾病预防控制中心,广东,广州,510080 |
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| 基金项目： | 国家科技重大传染病专项,国家高技术研究发展计划(863)资助项目,广东省科技攻关项目 |
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| 摘 要： | 目的:研制Ⅳ型登革病毒(DENV-4)非结构蛋白1(NS1)特异性的单克隆抗体(mAbs),并鉴定其特异性。方法:在表达具有良好抗原性的重组DENV4-NS1蛋白的基础上,用重组DENV4-NS1蛋白和灭活的DENV-4免疫BALB/c小鼠,取免疫小鼠的脾细胞与小鼠骨髓瘤细胞进行融合,用间接ELISA法筛选阳性的杂交瘤细胞,并结合免疫荧光测定法(IFA)和Western blot分析对mAb的特异性进行鉴定;通过竞争抑制试验对mAb识别的抗原位点进行分析。结果:获得15株特异性针对DENV4-NS1的mAbs,与其他3型DENV的NS1不产生交叉反应。mAb的Ig亚类测定均为IgG1。竞争抑制试验显示,15株mAbs存在2个不同识别抗原位点。结论:成功地获得特异性针对DENV4-NS1蛋白的mAbs,为进一步研究DENV4-NS1蛋白的结构与功能及研制DENV-4的临床诊断试剂奠定了基础。
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| 关 键 词： | 登革病毒 非结构蛋白1 单克隆抗体 |
Production and characterization of the monoclonal antibodies against nonstructural 1 protein of DENV-4 |
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| Ding XX,DI B,Che XY.Production and characterization of the monoclonal antibodies against nonstructural 1 protein of DENV-4[J].Journal of Cellular and Molecular Immunology,2011,27(7):777-779. |
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| Authors: | Ding Xi-xia DI Biao Che Xiao-yan |
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| Affiliation: | DING Xi-xia1,DI Biao2,CHE Xiao-yan1 1Center for Clinical Laboratory,Zhujiang Hospital,Southern Medical University,Guangzhou,510282,2Guangzhou Center for Disease Control and Prevention,510080,China |
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| Abstract: | AIM:To produce monoclonal antibodies(mAbs) against nonstructural 1 protein(NS1) of DENV-4 and characterization.METHODS:BALB/c mice were immunized with recombinant DENV4-NS1 protein and inactive DENV-4.Splenocytes of immunized mice were fused with myeloma cells(NS-1) to produce hybridoma cell lines,secreting anti-DENV4-NS1 protein mAbs.ELISA was used to identify the mAbs against NS1of DENV-4.Immnofluorescence assay(IFA) and Western blot analysis were applied to identify the specificity of mAbs.RESULTS:fiftee... |
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| Keywords: | dengue virus nonstructural 1 protein monoclonal antibody |
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