赭曲霉毒素A的抗原设计及在免疫分析中的应用

赭曲霉毒素A(Ochratoxin A，OTA)是一种重要的生物毒素，其广泛存在于谷物和其他植物性食品中。食品中的OTA残留进入人体后会对健康造成危害。为快速便捷地对OTA进行检测，本文以OTA为原料，设计合成了结构更加合理的新的OTA半抗原(OTA-H1)，制备了OTA的完全抗原(OTA-H1-BSA, OTA-H1-OVA)，并进行了小鼠免疫实验，建立了OTA间接竞争酶联免疫分析法(ic-ELISA)。采用核磁氢谱、核磁碳谱和高分辨质谱对OTA半抗原结构进行表征，结果表明半抗原结构正确；利用OTA-H1-BSA作为免疫抗原免疫Balb/c小鼠，小鼠血清效价为8 000，表明该免疫抗原对OTA具有一定的抑制作用；利用以OTA-H1-OVA作为包被抗原建立的间接竞争酶联免疫分析法检测小麦样品时，IC₅₀为0.04 ng/mL，灵敏度较原有方法提高了30倍。本方法可用于小麦样品中OTA的快速检测。

Antigen Design of Ochratoxin A and Its Application in Immunoassay

Ochratoxin A (Ochratoxin A, OTA) is an important biological toxin, which is widely found in grains and other plant foods. The OTA residues in food can cause health hazards after entering the human body. In order to detect OTA quickly and conveniently, this article uses OTA as the raw material, a new OTA hapten (OTA-H1) with reasonable structure was designed and synthesized using OTA as a raw material, and its structure was characterized by 1H NMR, 13C NMR and HRMS. The complete antigen (OTA-H1-BSA, OTA-H1-OVA) of OTA was obtained by activating OTA-H1 coupled with carrier protein. Using OTA-H1-BSA as the immune antigen to immunize Balb/c mice, the blood test was carried out and the results show that the serum titer of the mice is 8 000 and has a certain inhibitory effect on OTA. Using OTA-H1-OVA as the coating antigen, an indirect competitive enzyme-linked immunosorbent assay method (ic-ELISA) of OTA was established, the IC₅₀ of the method was 0.04 ng/mL, which was 30 times more sensitive than the original method. This method can be used for the detection of OTA in wheat samples.