| 抗鸡蛋卵类黏蛋白单克隆抗体的制备与鉴定 |
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| 引用本文： | 邹菊,张强,刘志刚. 抗鸡蛋卵类黏蛋白单克隆抗体的制备与鉴定[J]. 免疫学杂志, 2011, 0(3): 185-188 |
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| 作者姓名： | 邹菊 张强 刘志刚 |
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| 作者单位： | 深圳大学医学院过敏反应与免疫学研究所; |
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| 基金项目： | 深圳大学科研创新团队基金(200904); 广东省科技重点专项(2003A3080502); 深圳市深港创新圈项目(CXQ2008026); 深圳市科技计划(200929) |
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| 摘 要： | 目的制备和鉴定鼠抗鸡蛋主要过敏原卵类黏蛋白的单克隆抗体。方法利用鸡蛋卵类黏蛋白(ovm)为免疫原,免疫Balb/c小鼠,取免疫小鼠脾细胞与小鼠骨髓瘤NS-1细胞融合。采用半固体培养基法和有限稀释法相结合的方法快速筛选获得稳定分泌的特异性杂交瘤细胞。用杂交瘤细胞株诱生小鼠腹水,应用蛋白A亲和层析法进行抗体纯化。采用Ig类与亚类鉴定试剂盒鉴定该单克隆抗体的Ig亚型;通过间接ELISA、Western blotting鉴定该单克隆抗体的特性和交叉性。利用双抗体夹心ELISA法检测食品中的鸡蛋过敏原。结果获得4株可稳定分泌鼠抗卵类黏蛋白的单克隆抗体,分别命名为1G8,2H5,3A7,4G4,其Ig亚型均为IgG1。且4株单抗效价均在10-8以上。ELISA和Western blotting分析表明该4株单抗均能特异性识别卵类黏蛋白,并且建立双抗体夹心ELISA的方法可以准确的检测出食品中鸡蛋过敏原的存在。结论成功制备了4株高效价的鼠抗卵类黏蛋白的单克隆抗体,为建立卵类黏蛋白检测及纯化方法奠定了基础,也可为食品中鸡蛋过敏原的检出提供依据。
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| 关 键 词： | 卵类黏蛋白 单克隆抗体 特性鉴定 过敏原检测 |
Preparation and identification of monoclonal antibodies against allergen ovomucoid |
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| ZOU Ju,ZHANG Qiang,LIU Zhigang Allergy , Immunology Institute,Shenzhen University,Shenzhen ,China. Preparation and identification of monoclonal antibodies against allergen ovomucoid[J]. Immunological Journal, 2011, 0(3): 185-188 |
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| Authors: | ZOU Ju ZHANG Qiang LIU Zhigang Allergy Immunology Institute Shenzhen University Shenzhen China |
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| Affiliation: | ZOU Ju,ZHANG Qiang,LIU Zhigang Allergy and Immunology Institute,Shenzhen University,Shenzhen 518060,China |
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| Abstract: | To prepare monoclonal antibodies against allergen ovomucoid from chicken egg white and to characterize their properties,Balb/c mice were immunized with ovomucoid and the splenocytes of the immunized mice were fused with NS-1 myeloma cells by hybridoma technique.Four hybridoma cell lines secreting McAbs against ovomucoid allergen were obtained,which were determined as 1G8,2H5,3A7,4G4.The McAbs against ovomucoid allergen were purified using affinity chromatography on immobilized protein A and identified by th... |
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| Keywords: | Ovomucoid Monoclonal antibody Characterization Allergen test |
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