Flashner M, Ramsden H, Crane L J
Anal Biochem. 1983 Dec;135(2):340-4. doi: 10.1016/0003-2697(83)90693-0.
The chromatographic separation of four proteins, cytochrome c, alpha 1-acid glycoprotein, ovalbumin, and beta-lactoglobulin, was achieved on a 4.6 X 250-mm wide-pore polyethyleneimine (PEI)-silica gel column (5-micron particles, 330-A pore size) with essentially baseline resolution using a 20-min linear gradient from 0.025 M potassium phosphate, pH 6.80, to 0.50 M potassium phosphate, pH 6.80. The back pressure of this anion-exchange column was 1000 psi at a flow rate of 1.0 ml/min. Protein recoveries averaged over 95% and protein capacity exceeded 33 mg for a single protein. Isocratic elution (0.040 M potassium phosphate, pH 6.8; flow rate, 0.50 ml/min) of ovalbumin gave a column efficiency of 15,700 plates/m with a peak asymmetry factor of 1.27. Resolution of these same four proteins on a 4.6 X 50-mm PEI-silica gel column occurred within 2 min. Nucleoside monophosphates were separated on the short PEI-silica column within 1 min with 0.01 M potassium phosphate, pH 2.58, at a flow rate of 6 ml/min which generated a column back pressure of 2000 psi.
在一根4.6×250毫米的宽孔聚乙烯亚胺（PEI）-硅胶柱（5微米颗粒，孔径330埃）上实现了细胞色素c、α1-酸性糖蛋白、卵清蛋白和β-乳球蛋白这四种蛋白质的色谱分离，使用从0.025M磷酸钾（pH 6.80）到0.50M磷酸钾（pH 6.80）的20分钟线性梯度，基本达到基线分离度。该阴离子交换柱在流速为1.0毫升/分钟时背压为1000磅力/平方英寸。蛋白质回收率平均超过95%，单一蛋白质的载量超过33毫克。卵清蛋白等度洗脱（0.040M磷酸钾，pH 6.8；流速0.50毫升/分钟）时，柱效为15700理论塔板数/米，峰不对称因子为1.27。在一根4.6×50毫米的PEI-硅胶柱上，这四种相同的蛋白质在2分钟内实现了分离。核苷单磷酸在短的PEI-硅胶柱上，以0.01M磷酸钾（pH 2.58）、流速6毫升/分钟的条件在1分钟内实现了分离，此时柱背压为2000磅力/平方英寸。
