Millot M C, Hervé F, Sébille B
Laboratoire de Physico-Chimie des Biopolymères, Université Paris XII Val de Marne, Créteil, France.
J Chromatogr B Biomed Appl. 1995 Feb 3;664(1):55-67. doi: 10.1016/0378-4347(94)00352-6.
The retention behaviour of various amino acids, peptides and proteins on poly(vinylimidazole)-Cu(II) complexes supported on silica was investigated. Free amino acids and peptides containing one histidine and in some instances one additional tryptophan residue in their primary structure were found to elute from the supports only after addition of a competing complexing agent to the mobile phase. However, the results obtained the proteins containing metal binding groups suggested that, in addition to the presence of donor-acceptor interactions between the macromolecules and the immobilized metal, other additional (essentially ionic and/or hydrophobic) interactions took place between the proteins and the surrounding of the metal. When donor-acceptor interactions were predominant, proteins were strongly adsorbed on the stationary phase and their elution required the addition of a competing complexing agent in the mobile phase. However, when the binding between the proteins and the supports via donor-acceptor interactions was less favourable, proteins were eluted from the columns without the addition of a competing agent in the mobile phase. With respect to the binding of these proteins, ionic and/or hydrophobic interactions were no longer negligible during the chromatographic process and the retention of the macromolecules by the stationary phase depended on the elution conditions (ionic strength, pH, etc.). These supports were used in the fractionation of the three main genetic variants of desialylated alpha 1-acid glycoprotein.
研究了各种氨基酸、肽和蛋白质在硅胶负载的聚（乙烯基咪唑）-铜（II）配合物上的保留行为。发现游离氨基酸和在其一级结构中含有一个组氨酸且在某些情况下还含有一个额外色氨酸残基的肽，只有在向流动相中加入竞争络合剂后才会从载体上洗脱下来。然而，含有金属结合基团的蛋白质的研究结果表明，除了大分子与固定化金属之间存在供体-受体相互作用外，蛋白质与金属周围环境之间还发生了其他额外的（主要是离子和/或疏水）相互作用。当供体-受体相互作用占主导时，蛋白质强烈吸附在固定相上，其洗脱需要在流动相中加入竞争络合剂。然而，当蛋白质与载体通过供体-受体相互作用的结合不太有利时，蛋白质在不向流动相中加入竞争剂的情况下从柱中洗脱。关于这些蛋白质的结合，在色谱过程中离子和/或疏水相互作用不再可忽略不计，固定相对大分子的保留取决于洗脱条件（离子强度、pH等）。这些载体用于去唾液酸α1-酸性糖蛋白三种主要遗传变体的分离。
