文献类型：期刊文献

中文题名：黄曲霉毒素B_1完全抗原合成及鼠源多抗血清的制备

英文题名：Preparation of AflatoxinB_1 Complete Antigen and Mouse Polyclonal Antiserum

作者：魏继涛[1,2];胡延春[2];宁红梅[3];银梅[3];岳锋[1];贾文科[1];田献礼[3];王选年[1,3]

第一作者：魏继涛

机构：[1]新乡学院生物技术研究中心;[2]四川农业大学动物医学院;[3]河南科技学院动物科学学院

第一机构：新乡学院生命科学技术学院

年份：2012

卷号：28

期号：14

起止页码：44-49

中文期刊名：中国农学通报

外文期刊名：Chinese Agricultural Science Bulletin

收录：CSTPCD;;CSCD:【CSCD_E2011_2012】；

基金：河南省高校科技创新团队支持计划(2008IRTSTHN011)

语种：中文

中文关键词：黄曲霉毒素B1;完全抗原;多抗血清

外文关键词：Aflatoxin B1; complete antigen; polyclonal antiserum

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。
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 was good with half inhibitory concentration IC50 of 22.268 ng/mL,low cross-reactivity.The complete antigen of AFB1 was 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.