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.
出处
《中国农学通报》
CSCD
2012年第14期44-49,共6页
Chinese Agricultural Science Bulletin
基金
河南省高校科技创新团队支持计划(2008IRTSTHN011)
