Kopaciewicz W, Rounds M A, Regnier F E
J Chromatogr. 1985 Jan 18;318(2):157-72. doi: 10.1016/s0021-9673(01)90678-8.
Several anion-exchange stationary phases (based on polyethyleneimine-coated silica) were synthesized so as to vary in ligand density and hydrophobicity. These materials were first examined for hemoglobin-binding capacity and then evaluated chromatographically. Protein binding, retention and resolution increased concomitantly with ligand density. Ferritin (molecular weight 440,000) could not be eluted from the more highly-charged surfaces, but was desorbed from a low ligand density support. The above parameters also varied with the hydrophobic character of the stationary phase. Retention and resolution increased as more hydrophobic moieties were added. Data from a non-ionic hemoglobin-binding assay correlated reasonably well with anticipated matrix hydrophobicities. Possible explanations and applications of the observed phenomena are discussed.
合成了几种阴离子交换固定相（基于聚乙烯亚胺包覆的硅胶），使其在配体密度和疏水性方面有所不同。首先检测这些材料的血红蛋白结合能力，然后进行色谱评估。蛋白质结合、保留和分离度随配体密度的增加而同步提高。铁蛋白（分子量440,000）无法从电荷更高的表面洗脱，但可从低配体密度的载体上解吸。上述参数也随固定相的疏水特性而变化。随着添加更多疏水基团，保留和分离度增加。来自非离子血红蛋白结合测定的数据与预期的基质疏水性相当吻合。讨论了观察到的现象的可能解释和应用。
