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用于从实验室规模到生产规模的蛋白质和核酸纯化的新型高分辨率离子交换剂系列。

New family of high-resolution ion exchangers for protein and nucleic acid purifications from laboratory to process scales.

作者信息

Dion D M, O'Connor K, Phillips D, Vella G J, Warren W

机构信息

Waters Chromatography, Division of Millipore Corporation, Milford, MA 01757.

出版信息

J Chromatogr. 1990 Dec 28;535(1-2):127-45. doi: 10.1016/s0021-9673(01)88939-1.

Abstract

A new family of polymer-based ion exchangers was tested for the purification of acidic and basic proteins on both the analytical and preparative scales. Protein-Pak HR series packings are available as strong cation (SP) and weak anion (DEAE) exchangers, allowing the development of a purification method regardless of the isoelectric point of protein. Three particle sizes, 8, 15 and 40 microns, are offered in scalable Advanced Purification (AP) glass columns or as bulk packings. The lower back pressures of the 15- and 40-microns packings compared to that of the 8-microns material allow rapid throughput of large volumes without exceeding the pressure limitations of the resin or the column. The capacity of the AP1 (100 mm x 10 mm) glass columns, containing these ion-exchange packings, is comparable to other ion-exchange columns. The resolution of mouse serum, plasmids, and a standard protein mixture was demonstrated and compared with the results obtained with other resin-based ion exchangers of similar particle size. Proteins were purified without significant loss of biological activity or mass.

摘要

测试了一种新型的基于聚合物的离子交换剂家族,用于在分析和制备规模上纯化酸性和碱性蛋白质。Protein-Pak HR系列填料有强阳离子(SP)和弱阴离子(DEAE)交换剂可供选择,无论蛋白质的等电点如何,都能开发出一种纯化方法。提供三种粒径,分别为8、15和40微米,可用于可扩展的高级纯化(AP)玻璃柱或作为散装填料。与8微米材料相比,15微米和40微米填料的背压低,可在不超过树脂或柱子压力限制的情况下实现大量样品的快速通量。含有这些离子交换填料的AP1(100毫米×10毫米)玻璃柱的容量与其他离子交换柱相当。展示了小鼠血清、质粒和标准蛋白质混合物的分离效果,并与使用其他类似粒径的基于树脂的离子交换剂获得的结果进行了比较。蛋白质纯化过程中生物活性和质量没有明显损失。

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