赭曲霉毒素A人工抗原的合成与鉴定

Synthesis and identification of ochratoxin A artificial immunogen

采用N-羟琥珀酰亚胺酯(N-hydroxysuccinimide NHS)法将赭曲霉毒素A(OTA)与牛血清白蛋白(BSA)偶联制备了免疫抗原OTA-BSA,采用1-乙基-3(3-二甲基氨基丙基)碳二亚胺(EDC)法将赭曲霉毒素A(OTA)与卵清白蛋白(OVA)偶联制备了包被抗原OTA-OVA。紫外扫描和SDS-PAGE凝胶电泳结果初步表明偶联成功。用OTA-BSA分10μg/只和50μg/只两个剂量分别免疫BALB/C小鼠,获得了高效价的特异性多克隆抗血清,效价可达1∶104,抑制效价达14.7ng,证明偶联成功,并显示低剂量免疫可以提高小鼠pAb的敏感性。本研究为OTA单抗的制备及其免疫学分析方法的建立奠定了基础。

OTA was linked to the carrier proteins BSA with NHS method to prepare the immunogen OTA-BSA,and linked to the carrier proteins OVA with EDC method to prepare the coating antigen OTA-OVA.UV scanning spectrum and SDS-PAGE preliminarily proved that the hapten OTA was coupled successfully to the carrier protein.The specific polyclonal antiserum with high titre were obtained by immunizing BALB/C mice with OTA-BSA of 10μg and 50μg each respectively,with the titre 1∶104 and the inhibition titer 14.7ng.These results indicated that OTA was successfully linked to the carrier proteins,and showed that a low dose of immunization can increase the sensitivity of pAb in the mouce.This study lay the foundation for further gain monoclonal anti-OTA and its immunological assay.