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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (14): 44-49.doi: 10.11924/j.issn.1000-6850.2011-3182

所属专题： 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

黄曲霉毒素B1完全抗原合成及鼠源多抗血清的制备

魏继涛 胡延春 宁红梅 银梅 岳锋 贾文科 田献礼 王选年   

  • 收稿日期:2011-10-31 修回日期:2012-03-07 出版日期:2012-05-15 发布日期:2012-05-15
  • 基金资助:

    河南省高校科技创新团队支持计划

Preparation of AflatoxinB1 Complete Antigen and Mouse Polyclonal Antiserum

  • Received:2011-10-31 Revised:2012-03-07 Online:2012-05-15 Published:2012-05-15

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摘要：

为了合成黄曲霉毒素B1(AFB1)完全抗原，制备鼠源AFB1多克隆抗体血清。通过琥珀酰亚胺酯法在黄曲霉毒素B1(AFB1)分子上引入羧基，生成黄曲霉毒素B1肟(AFB1O)。用EDC法和DCC法合成AFB1-BSA和AFB1-OVA。采用薄层层析技术、紫外光谱技术(Uv)、SDS-PAGE鉴定完全抗原的合成。通过免疫BALB/c小鼠，获得鼠原多克隆血清，采用间接ELISA方法测定多抗血清的免疫效价，用竞争ELISA检测多克隆血清的敏感性和特异性。结果表明：免疫小鼠血清效价都在1:3200以上，其中6号鼠效价最高，到达1.28×10-4，敏感性好，半数抑制浓度IC50为22.268 ng/mL，交叉反应率低。本研究成功制备了AFB1完全抗原，获得了敏感的AFB1多克隆抗体血清，为以后制备AFB1单克隆抗体及免疫学快速检测方法奠定了基础。

关键词: 植烟土壤, 植烟土壤, 养分, 评价, 黔江区

Abstract:

The complete antigen of AFB1 was synthesized. The mouse polyclonal antiserum was prepared by immunizing the BALB/c mice with the complete antigen. The carboxyl group was introduced in the Aflatoxin B1 (AFB1) molecule by Succinimide ester method. AFB1O was produced. AFB1-BSA and AFB1-OVA were synthesized by EDC and DCC methods. Thin layer chromatography technology, Uv spectroscopy, SDS-PAGE and immunological methods were used to identify the complete antigen. BALB/c mice were immunized with AFB1-BSA. The titer, sensibility and specificity of the antiserum was determined by indirect ELISA and competitive ELISA. The results showed that the antiserum titer of all the BALB/c mice were over 1:3200. The highest No.6 mouse reached 1.28×10-4 and the sensitivity is good with half inhibitory concentration IC50 of 22.268 ng/mL, low cross-reactivity. The complete antigen of AFB1was synthesized successfully. The sensitive polyclonal antiserum of AFB1 has been gained in this study, which laid a foundation for the preparation of monoclonal antibodies and the establishment of a rapid detection method against AFB1.