T-2毒素人工抗原的制备

Preparation of T-2 Toxin Antigen

在蒸气浴条件下,T-2毒素与琥珀酸酐(HS)反应合成了T-2HS,薄层层析显示,目标半抗原合成成功;然后通过碳二亚胺法将半抗原与载体蛋白偶联制备人工抗原,采用紫外扫描及SDS-PAGE鉴定。结果显示,T-2HS能与牛血清白蛋白(BSA)和卵清蛋白(OVA)结合生成T-2毒素抗原。紫外扫描分析表明,T-2毒素与牛血清白蛋白的偶联比为6.66∶1、与卵清蛋白的偶联比为10.11∶1,表明此完全抗原能够用于免疫动物。为进一步制备T-2毒素抗体奠定了基础。

First,T-2 toxin reacted with succinic anhydride in steam-bath to synthesize T-2 hemisuccinate（T-2HS）.Thin layer chromatography analysis showed that the objective hapten was obtained successfully.Then,antigen was prepared by conjugation of hapten with carrier protein by carbodiimide（EDPC） method and was identified by UV spectroscopy and SDS-PAGE.The result showed that T-2 toxin could conjugate with BSA or OVA to produce T-2 toxin antigen.UV Spectrum analysis showed that the conjugation ratio of T-2 toxin to BSA was 6.66∶1 and that to OVA was 10.11∶1,indicating this antigen could be used to immunize animals.It provided basis for preparing T-2 toxin antibody.