伏马菌素B_1人工抗原的制备及其鉴定

Preparation and Identification of the Fumonisin B_1 Artificial Antigen

为制备人工抗原FB1-OVA,采用戊二醛一步法将小分子半抗原-伏马菌素B1(FB1)与载体蛋白OVA进行偶联。分别采用聚丙烯酰胺凝胶电泳法(SDS-PAGE)、紫外扫描法(UV)、免疫芯片法和质谱法(MS)来鉴定FB1与OVA的偶联效果。结果显示,凝胶电泳图中FB1-OVA的电泳条带迁移率比OVA的迁移率小,紫外扫描图谱FB1-OVA的波型与OVA的波型发生了明显的变化,免疫芯片检测可见偶联物与FB1McAb发生了特异性反应,初步证明偶联成功。质谱法检测得出FB1-OVA和OVA的相对分子质量分别是51 928.612和44 265.718,可计算出偶联比为10.6∶1,通过测定,偶联物的蛋白质浓度为0.99 mg/mL。

The hapten fumonisin B1 （ FB1 ） was coupled with carrier proteins ovalbumin （OVA） by one - step glutaraldehyde method to prepare the artificial antigen （FB1 -OVA）. Sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS -PAGE）, ultraviolet spectroscopy （UV）, mass spectrum （MS） and immunoassay method were adopted respectively to confirm the conjugate effect of FB1 - OVA. The results indicated that the FB1 - OVA electro- phoresis banding migration rate was lower than the mobility of OVA in gel electrophoresis figure; the wave pattern of FB1 - OVA and OVA had changed obviously. The immunoassay results showed that the coupling and FB; McAb were specificity reacted. It could preliminary estimate the coupling was obtained successfully. The molecular weights of FB1 - OVA and OVA were respectively 51 928. 612 and 44 265. 718 by MS and the conjugate ratio was 10.6： 1, and the concentration of coupling was 0.99 mg/mL.