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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (11): 78-81.

• 食品 营养 检测 安全 • 上一篇    下一篇

叶酸人工抗原合成及免疫抗血清的鉴定

张会   

  • 收稿日期:2010-09-08 修回日期:2010-11-17 出版日期:2011-05-15 发布日期:2011-05-15

Preparation of Folic acid artificial antigen and identification of antiserum

  • Received:2010-09-08 Revised:2010-11-17 Online:2011-05-15 Published:2011-05-15

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摘要：

叶酸是人体必需的营养物质，叶酸的缺乏可以引起神经管畸形等多种疾病，建立叶酸的快速检测方法对营养强化食品的质控以及临床研究具有重要意义。本实验通过活化酯法合成叶酸-牛血清白蛋白（FA-BSA）人工免疫原，通过混合酸酐法制备叶酸-卵清蛋白（OVA）包被原。通过紫外扫描和免疫动物进行人工抗原的合成鉴定。根据BSA、OVA的含量以及人工抗原、BSA、OVA及叶酸（FA）的紫外吸收曲线在280nm处的摩尔吸收值，计算出叶酸与BSA的偶联比为1：3.84， 叶酸与OVA的偶联比为4.70。利用合成的叶酸人工抗原免疫小鼠，三免后眼眶内眦静脉采血，采用间接ELISA方法测定免疫小鼠多抗血清效价在128,000以上，叶酸对抗血清的IC50为14.82ng/ml；由此表明叶酸人工免疫原合成成功，为进一步研究叶酸单克隆抗体奠定基础。

关键词: 磷石膏, 磷石膏, 农业, 土壤, 生物降解, 安全

Abstract:

Folic acid, as the necessary nutrition of human, can defense many diseases, such as NDFS, so building up a rapid test method of folic acid is important to quality control of food or research in clinic. The Folic acid hapten was synthesized by Williamson synthesis method. immunogen and coating conjugate have been synthesized. Balb/c Mice were immunized with the folic acid conjugated with bovie serum avumin. The antiserum titer determined by indirect ELISA was more than 1.28×105，Using antibodies, the indirect competivive ELISA was established which was suitable for detecting FA: the IC50 was 14.28ng/mL.