Contado Catia, Bregola Letizia, Dondi Francesco
Department of Chemistry, University of Ferrara, Via L. Borsari, 46, I-44100 Ferrara, Italy.
J Chromatogr A. 2007 Oct 26;1169(1-2):158-74. doi: 10.1016/j.chroma.2007.08.069. Epub 2007 Sep 4.
The amount of immunoglobulin A (IgA) adsorbed on the surface of two different samples of polystyrene (PS) microbeads was evaluated using differential sedimentation field flow fractionation (SdFFF) analyses. For the first time, the SdFFF separations obtained by using, as mobile phase, solutions common to many biochemical procedures and applications have been compared and discussed. Good separation results were achieved in the different carriers, and the SdFFF gave equivalent mass per particle values in all carriers provided that the pH and ionic strength conditions of the eluents were well controlled. The IgA adsorption process onto PS occurred by maintaining unaltered the capacity of the PS-IgA substrate to selectively recognize anti-IgA (aIgA), as proven by elution of the ternary complex PS-IgA-aIgA and from the monitored lack of reaction when the PS-IgA was placed in contact with aIgE.
使用差示沉降场流分级法(SdFFF)分析评估了吸附在两种不同聚苯乙烯(PS)微珠样品表面的免疫球蛋白A(IgA)的量。首次对以许多生化程序和应用中常见的溶液作为流动相获得的SdFFF分离结果进行了比较和讨论。在不同的载体中均取得了良好的分离效果,并且只要洗脱液的pH值和离子强度条件得到很好的控制,SdFFF在所有载体中给出的每个颗粒的等效质量值都是相同的。PS上IgA的吸附过程通过保持PS-IgA底物选择性识别抗IgA(aIgA)的能力不变来实现,三元复合物PS-IgA-aIgA的洗脱以及将PS-IgA与aIgE接触时监测到的无反应情况证明了这一点。