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中国农学通报

中国农学通报 ›› 2014, Vol. 30 ›› Issue (17): 26-30.doi: 10.11924/j.issn.1000-6850.2013-3318

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

卡那霉素人工抗原的制备及鉴定

王爱萍 栗宁 祁艳华 段倩倩 郑伟 王新洲 裴艳艳 周景明   

  • 收稿日期:2013-12-19 修回日期:2014-03-19 出版日期:2014-06-14 发布日期:2014-06-14
  • 基金资助:
    国家科技支撑计划子课题项目“食品中重要危害物抗体库的建立及产品研发”(20111BAK10B01-17)；河南省重点科技攻关项目“噬菌体展示技术淘选赭曲霉毒素模拟表位研究”(122102310629)。

Synthesis and Identification of Kanamycin Complete Antigen and Preparation of Polyclonal Antibody Against Kanamycin

  • Received:2013-12-19 Revised:2014-03-19 Online:2014-06-14 Published:2014-06-14

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摘要： 为了人工制备卡那霉素完全抗原。将卡那霉素标准品与牛血清白蛋白(BSA)或卵清白蛋白(OVA)偶联，制备人工抗原。利用两种不同的偶联方法：碳二亚胺法(EDC)和戊二醛法(GDA)，分别制备出KAN-BSA（作为免疫原），KAN-GDA-OVA（作为包被原）。利用SDS-PAGE凝胶电泳鉴定和动物免疫试验等，鉴定了完全抗原偶联情况。结果显示：制备的卡那霉素完全抗原能刺激小鼠机体产生相应抗体，且抗血清效价均达到1:2.56×104，其中一号小鼠的IC50值为22.28 ng/mL，说明完全抗原免疫原性良好，抗原制备成功。

关键词: 风险应对, 风险应对

Abstract: The purpose of this experiment was preparing kanamycin completely artificial antigens. Firstly, kanamycin standard was reacted with bovine serum albumin (BSA) or ovalbumin (OVA) to prepare kanamycin artificial antigens. Two different coupling methods carbodiimide (EDC) and glutaraldehyde (GDA) were respectively prepared KAN-BSA (as an immunogen) and KAN-GDA-OVA (as the coating antigen). Both of the SDS-PAGE gel electrophoresis and the animal immunization experiment identified the antigen-coupled case. The results showed that, the prepared kanamycin complete antigen could stimulate the bodies to produce antibodies in mice, and the antiserum titer reached 1:2.56×104, The IC50 of the mice was 22.28 ng/mL. The results illustrated the immunogenicity of the complete antigens were good and the preparations of the antigens were successful.