Jankowski W J, von Muenchhausen W, Sulkowski E, Carter W A
Biochemistry. 1976 Nov 16;15(23):5182-7. doi: 10.1021/bi00668a036.
Blue Dextran (Cibacron Blue F3GA-dextran) was immobilized on cyanogen bromide activated agarose and used as a ligand for human fibroblast and leukocyte interferons in a solvent of phosphate-buffered (pH 7.4), physiological saline (0.15 M NaCl). Fibroblast interferon binds completely and is not displaced from the column by an increase in ionic strength of the solvent (1.0 M NaCl); it can be, however, recovered with ethylene glycol, indicating the hydrophobic nature of interaction. Leukocyte interferon also binds to Blue Dextran-agarose but it can be recovered simply by an increase in the ionic strength of the solvent, indicating primarily the electrostatic nature of binding. Attempts to displace both interferons selectively with nucleosides and aromatic amino acids were unsuccessful. When Cibacron Blue F3GA is immobilized directly to agarose matrix or via molecular arm, the strength of binding of fibroblast interfern is significantly decreased, although ethylene glycol is still required for its displacement from the column. Leukocyte interferon, by contrast, does not bind at all under the same solvent conditions; it does bind when the pH value of the solvent is in the range 3-5 i.e., below its isoelectric point. Human fibroblast interferon binds completely to: aminobenzene, aminonaphthalene, and aminoanthracene, all immobilized on agarose, and it can be recovered with ethylene glycol. In contrast, human leukocyte interferon does not bind to benzene-agarose; it is retarded on naphthalene-agarose and completely retained on an anthracene-agarose column. All data point to a higher intrinsic hydrophobicity of human fibroblast interferon vis-á-vis human leukocyte interferon. Selective binding of human fibroblast interferon of Cibacron Blue F3GA-agarose results in a significant purification, about 800-fold.
蓝色葡聚糖（汽巴克隆蓝F3GA - 葡聚糖）被固定在溴化氰活化的琼脂糖上，并在磷酸盐缓冲（pH 7.4）的生理盐水（0.15M NaCl）溶剂中用作人成纤维细胞和白细胞干扰素的配体。成纤维细胞干扰素完全结合，并且不会因溶剂离子强度增加（1.0M NaCl）而从柱上被置换下来；然而，它可以用乙二醇回收，这表明相互作用具有疏水性。白细胞干扰素也与蓝色葡聚糖 - 琼脂糖结合，但只需增加溶剂的离子强度就可以回收，这主要表明结合具有静电性质。用核苷和芳香族氨基酸选择性置换这两种干扰素的尝试均未成功。当汽巴克隆蓝F3GA直接固定在琼脂糖基质上或通过分子臂固定时，成纤维细胞干扰素的结合强度显著降低，尽管仍需要乙二醇将其从柱上置换下来。相比之下，白细胞干扰素在相同的溶剂条件下根本不结合；当溶剂的pH值在3 - 5范围内，即低于其等电点时，它才会结合。人成纤维细胞干扰素完全结合到固定在琼脂糖上的苯胺、氨基萘和氨基蒽上，并且可以用乙二醇回收。相比之下，人白细胞干扰素不与苯 - 琼脂糖结合；它在萘 - 琼脂糖上迁移受阻，并完全保留在蒽 - 琼脂糖柱上。所有数据表明，相对于人白细胞干扰素，人成纤维细胞干扰素具有更高的内在疏水性。人成纤维细胞干扰素与汽巴克隆蓝F3GA - 琼脂糖的选择性结合导致了显著的纯化，约800倍。
