Influence of hapten density on immunogenicity for anti-ciprofloxacin antibody production in mice
Influence of hapten density on immunogenicity for anti-ciprofloxacin antibody production in mice
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DOI：
10.5582/bst.2012.v6.2.52
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发表时间：
2012-01-01
影响因子：
5.5
通讯作者：
Yang, Xianle
中科院分区：
文献类型：
--
作者：
Hu, Kun;Huang, Xuanyun;Yang, Xianle
为了产生针对小分子的抗体，有必要将它们作为半抗原偶联到大载体如蛋白质上。然而，缀合物的免疫原性通常与半抗原-蛋白质比率不具有线性相关性，这可能导致所需抗体的特性的较大变化。采用碳二亚胺法将环丙沙星（CPFX）与牛血清白蛋白（BSA）以5种不同的比例偶联。通过分光光度法和电泳法对偶联物进行了定性表征。采用质谱法和三硝基苯磺酸法对偶联物的密度进行定量测定。结果，获得了具有各种半抗原密度（每个载体蛋白21-30个分子）的CPFX-BSA缀合物。免疫小鼠后，ELISA检测表明，随着半抗原浓度的增加，抗血清效价逐渐升高。从小鼠获得的抗体显示出对CPFX的高敏感性。这些结果揭示了半抗原密度和免疫原性之间的关系以及用于免疫目的的优化缀合方法。
To generate antibodies against small molecules, it is necessary to couple them as haptens to large carriers such as proteins. However, the immunogenicity of the conjugates usually has no linear correlation with the hapten-protein ratio, which may lead to large variations in the character of the desired antibodies. In the present study, ciprofloxacin (CPFX) was coupled to bovine serum albumin (BSA) in five different proportions using a modified carbodiimide method. The conjugates were characterized qualitatively by spectrophotometric absorption and electrophoresis methods. Mass spectrometry and the trinitrobenzene sulfonic acid method were adopted to assay the density of conjugates quantitatively. As a result, CPFX-BSA conjugates with various hapten densities (21-30 molecules per carrier protein) were obtained. After immunization in mice, ELISA tests showed that the antisera titer increased gradually with the increase of hapten density. The antibody obtained from the mice showed high sensitivity toward CPFX. These results revealed the relationship between hapten density and immunogenicity as well as an optimized conjugation approach for immunization purposes.