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1:25600的抗体,间接竞争ELISA测定抗体IC50值为39ng/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.