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Ⅰ型登革病毒NS1蛋白特异性血清型单克隆抗体的制备及鉴定
引用本文:徐华,郝卫,潘玉先,丘立文,王压娣,何丽娟,狄飚,车小燕.Ⅰ型登革病毒NS1蛋白特异性血清型单克隆抗体的制备及鉴定[J].广东医学,2006,27(3):308-310.
作者姓名:徐华  郝卫  潘玉先  丘立文  王压娣  何丽娟  狄飚  车小燕
作者单位:1. 南方医科大学珠江医院中心实验室,广州,510282
2. 广东省广州市疾病预防控制中心,510080
摘    要:目的研制Ⅰ型登革病毒(DEN1)非结构蛋白1(NS1)蛋白特异性单克隆抗体,鉴定其特异性。方法以具有良好免疫原性的DEN1重组NS1蛋白、DEN1全病毒及两者混合免疫共3种免疫方案,分别免疫Balb/c小鼠,取其脾细胞与小鼠骨髓瘤细胞融合,杂交瘤细胞经间接ELISA法双筛I、FA和Western Blot进行mAbs的类型及亚类、特异性等鉴定。结果采用3种免疫方案免疫Balb/c小鼠,获得9株抗NS1蛋白的mAb,其亚类测定一株为IgG2a,另外8株为IgG1。这些mAbs与DEN1重组NS1蛋白和DEN1全病毒均结合,而且特异性好,仅1株杂交瘤分泌的单抗和其余3型DEN有交叉反应。结论成功获得了特异性针对DEN病毒NS1蛋白的特异性血清型mAb,将为进一步研究NS1蛋白的结构和功能及临床诊断试剂研发奠定基础。

关 键 词:登革病毒  非结构蛋白  单克隆抗体
收稿时间:2005-09-16
修稿时间:2005年9月16日

Preparation and characterization of monoclonal antibodies against DEN1 nonstructural 1 protein
XU Hua, HAO Wei, PAN Yu-xian.Preparation and characterization of monoclonal antibodies against DEN1 nonstructural 1 protein[J].Guangdong Medical Journal,2006,27(3):308-310.
Authors:XU Hua  HAO Wei  PAN Yu-xian
Institution:Central Laboratory of Zhujiang Hospital, Nanfang Medical University, Guangzhou 510282, China
Abstract:Objective To prepare monoclonal antibodies against dengue serotype 1 virus(DEN1) nonstructual 1 protein and to perform immunological identification to the antibodies.Methods 9 Balb/c mice were divided into three groups: immunized by recombinant DEN1 nonstructual 1 protein of high immunogenicity,immunized by inactive DEN1 virus,or received several booster of recombinant DEN1 nonstructual 1 protein after two injections of inactive DEN1 virus.Splenocytes of immunized mice were fused with myeloma cells NS-1 to produce hybridoma cell line,ecreting anti-DEN nonstructual 1 protein antibodies.Enzyme-linked immunosorbent assay(ELISA),immnofluorescence assay(IFA) and Western blot analysis were applied to identify specificity of antibodies.Results Nine strains of hybridoma cell lines steadily ecrering antibodies of nonstructual 1 protein were obtained.These antibodies had characteristics of specific binding to DEN and recombinant nonstructual 1 protein.Among them,one strain was identified as IgG2a isotype and other eight were all IgG1.Eight out of nine strains were specific to DEN1 without detectable cross-reactivity with other three serotypes of dengue virus.Conclusion Monoclonal antibodies against DEN1 nonstructual 1 protein with high activity and specificity had been successfully established.These results could provide a potential value for vaccine researches and early diagnosis of dengue virus infection.
Keywords:Dengue virus Nonstructual 1 protein Monoclonal antibodies
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