黄曲霉毒素B1人工抗原的合成及鉴定

Synthesis and Identification of Aflatoxin B1 Artificial Antigen

摘要:

采用衍生化在黄曲霉毒素B1(aflatoxin B1，AFB1）上引入羧基合成AFB1羧甲基活化物，通过N-羟琥珀酰亚胺酯(N-hydroxy-succinimide NHS）法将AFB1O与牛血清白蛋白(BSA)偶联，制备黄曲霉毒素B1完全抗原AFB1-BSA。ECI-MS和紫外光谱法的鉴定结果表明目标半抗原合成成功。结合紫外分光光度法和回归方程，分别测得不同浓度的半抗原和蛋白质线性曲线为：Y=0.1440X+0.0103，R2=0.9986；Y=0.0059X+0.0808，R2=0.9889。偶联物中半抗原和蛋白质的浓度分别为186.32 μg/mL、6127.46 μg/mL，即求得抗原分子结合比为5.13:1，从而为制备抗AFB1抗体奠定基础。

Abstract:

Carboxyl group was introduced to Aflatoxin B1 (aflatoxin B1, AFB1) by derivatization method to synthesis the hapten (AFB1 carboxymethyl activator). And the AFB1O was coupled with BSA by using N-hydroxy-succinimide method to prepare the complete antigen of AFB1. The results of ECI-MS and ultraviolet spectroscopy showed that the target hapten was successfully synthesized. Through combined with ultraviolet spectrophotometry and regression equation, the standard curves of the different concentration hapten and BSA were abtained as follow: y=0.1440x+0.0103 (R2 =0.9986) and y=0.0059x+0.0808 (R2 =0.9889) respectively. The concentrations of AFB1O and BSA in adduct were 186.32 μg/mlL and 6127.46 μg/mL respectively, and the molar ration was 5.13:1.