黄曲霉毒素G族人工抗原的合成与免疫效果研究

Synthesis of aflatoxin G class artificial antigen

以黄曲霉毒素(Aflatoxin G1,AFG1)的半缩醛(Aflatoxin G2a,AFG2a)为中间物,通过还原烷基法,使黄曲霉毒素G1在8、9羟基上与牛血清白蛋白(BSA)实现偶联,得到偶联物。对偶联物进行紫外全波长扫描显示,其特征吸收峰与BSA和AFG1的特征吸收峰有明显差异。经完全透析后检测偶联物与BSA荧光强度差异,均表明BSA与黄曲霉毒素G1偶联,AFG1-BSA物质的量结合比为3.2∶1。以合成人工抗原免疫小鼠,用间接非竞争ELISA测得抗血清中抗体的效价可达1∶128 000,间接竞争ELISA测定抗体IC50为14.7 ng/mL。为下一步研制黄曲霉毒素G族单克隆抗体提供了关键试剂,为研究建立黄曲霉毒素G族免疫速测技术奠定了基础。

The hemiacetal of aflatoxin G1（AFG1） was conjugated to BSA at the 8,9-hydroxy groups through the reducing alkylation reaction to yield the artificial antigen for aflatoxin G class.The UV spectra and fluorescence intensity of the conjugated AFG1-BSA were obviously different from those of BSA and aflatoxin G1,attesting to the successful preparation of conjugated AFG1-BSA.The coupling ratio of AFG1 to BSA was 3.2∶1 in the conjugate.After being immunized with AFG1-BSA for 12 weeks,the titer of the antiserum could reach 1∶128 000 and the IC50 was 14.7 ng/mL for the aflatoxin G1.In this paper,the prospects for the development of a fast method for testing aflatoxin G class were referred to.