测定克喘素免疫偶合物分子结合摩尔比的紫外-可见分光光度法

Determining the Molecular Coupling Ratio of Clenbuterol Immunoconjugates by Ultraviolet-Visible Spectrophotometry

以牛血清白蛋白 (BSA)、卵清白蛋白 (OVA)为载体,采用重氮法,制出克喘素 (CL)的两种免疫偶合物免疫原CL-BSA和包被抗原CL -OVA ;通过紫外光谱定性证明偶合物偶联成功 ,通过可见光谱法定量测定偶合物的分子结合摩尔比 ,结果表明CL -BSA与CL-OVA的分子结合摩尔比分别为13.19∶1、7.53∶1,方法简便快捷 。

Clenbuterol (α[t- butylaminomethyl]- 4- amino- 3,5- dichlorobenzyl alcohol) can be successfully coupled to albumin by the diazonium method, as verified by their UV spectra. With bovine serum albumin(BSA) and ovalbumin(OVA) as carriers, two clenbuterol(CL) immunoconjugates have been thus prepared, namely, the immunogen of CL-BSA and the coating antigen of CL-OVA. The molecular coupling ratios of CL-BSA and CL-OVA have been determined to be 13.19∶1 and 7.53∶1, respectively, by the visible spectrometry. The method is simple, rapid and highly sensitive.