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聚乙二醇键合相已用于高效疏水作用色谱分离活性蛋白质。本文中,我们进一步研究了聚乙二醇的分子量和硅胶孔径对键合相覆盖率和蛋白质分离效能的影响。实验结果表明,较大孔径的键合相具有高的覆盖率和好的分离效能。分子量小的聚乙二醇键合相适合于分离疏水性强的蛋白质;反之,分子量大的键合相则对亲水性蛋白质有好的分离效果。其中PEG-1500键合相(50nm)具最佳分离效能。蛋白质在此柱上可得90％以上的活性回收率。2mg以上的蛋白质样品可一次注入100×5mm I.D.分析柱上而不影响分辨率和保留时间。
Polyethylene glycol bonded phases have been used for the efficient hydrophobic interaction chromatography for the separation of active proteins. In this paper, we further investigate the molecular weight of polyethylene glycol and silica gel pore size on the bonding phase coverage and protein separation efficiency. The experimental results show that the larger-diameter bonded phase has high coverage and good separation performance. Low molecular weight polyethylene glycol bonds are suitable for the separation of highly hydrophobic proteins; conversely, high molecular weight bonded phases have a good separation of hydrophilic proteins. PEG-1500 bonded phase (50nm) with the best separation performance. The protein has an activity recovery of more than 90% on this column. A protein sample of 2 mg or more can be injected into a 100 × 5 mm I.D. analytical column once without affecting resolution and retention time.