Rezeli Melinda, Kilár Ferenc, Hjertén Stellan
Institute of Chemistry, Faculty of Science, University of Pécs, H-7624 Pécs, Hungary.
J Chromatogr A. 2006 Mar 17;1109(1):100-2. doi: 10.1016/j.chroma.2005.11.076. Epub 2005 Dec 15.
The introductions of the continuous beds, now often called monoliths [S. Hjertén, J.-L. Liao, R. Zhang, J. Chromatogr. 473 (1989), 273-275] and the artificial, highly selective gel antibodies against antigens as large as proteins, viruses and cells [J.-L. Liao, Y. Wang, S. Hjertén, Chromatographia 42 (1996), 259-262] were breakthroughs in the design of chromatographic beds. This paper deals with a combination of these two methods, i.e., the artificial gel antibodies have been synthesized in the monolithic mode. As antigen we have used human hemoglobin. A comparison of the ion-exchange chromatograms of the eluates from the monolithic columns shows that the monolith prepared in the presence of hemoglobin adsorbed this protein, but not the other proteins in the sample (ribonuclease A and cytochrome c), i.e., this monolith was selective for hemoglobin, whereas the blank column (prepared in the absence of hemoglobin) had no selective properties, since none of the applied proteins were adsorbed. The diameter of the column was 6mm, but the same approach to synthesize a monolithic selective bed can very likely also be used for capillaries and microchips.
连续床（现常称为整体柱 [S. Hjertén, J.-L. Liao, R. Zhang, J. Chromatogr. 473 (1989), 273 - 275]）以及针对诸如蛋白质、病毒和细胞等大分子抗原的人工高选择性凝胶抗体 [J.-L. Liao, Y. Wang, S. Hjertén, Chromatographia 42 (1996), 259 - 262] 的引入是色谱柱设计方面的突破。本文探讨这两种方法的结合，即人工凝胶抗体以整体柱模式合成。我们使用人血红蛋白作为抗原。对整体柱洗脱液的离子交换色谱图进行比较表明，在血红蛋白存在下制备的整体柱吸附了该蛋白质，但未吸附样品中的其他蛋白质（核糖核酸酶A和细胞色素c），即该整体柱对血红蛋白具有选择性，而空白柱（在无血红蛋白的情况下制备）没有选择性，因为所施加的蛋白质均未被吸附。柱的直径为6毫米，但合成整体选择性柱的相同方法很可能也可用于毛细管和微芯片。
