三聚氰胺完全抗原的制备与表征

Preparation and Characterization of Complete Antigen of Melamine

采用戊二醛法将三聚氰胺(Melamine,MEL)偶联到牛血清白蛋白(BSA)上制备完全抗原MEL-BSA,并用紫外扫描、红外扫描及SDS-PAGE电泳试验进行了鉴定。结果表明,半抗原MEL和BSA偶联成功,紫外光谱分析法计算得其偶联比为7.9∶1。通过免疫动物获得效价为1∶25 600的抗体,间接竞争ELISA测定抗体IC50值为39 ng/mL,与三聚氰酸及其他几种常用抗菌素的交叉反应率均<0.2%,为下一步研制三聚氰氨单克隆抗体,建立三聚氰氨免疫速测技术奠定了基础。

In order to effectively detect melamine （MEL） residues,a complete antigen of MEL was developed by glutaraldehyde synthesis methods. MEL was coupled with bovine serum albumin （BSA） to obtain a complete antigen of MEL- BSA. Infrared and ultraviolet scanning spectra and SDS-PAGE analysis confirmed that the complete antigen of MEL-BSA had been successfully coupled by the method. The coupling molar ratios were calculated to be 7.9：1 based on UV spectroscopy analysis. The titer of the antibodies secreted by immunized an- imals was 1 ： 25 600, the lCso was 39 ng/mL for melamine and the cross-reactivity with eyanuric acid and other antibiotics was less than 0.2%. Finally, the probability for development of the fast immunization testing method for melamine residue was discussed.