Cabrera Rosa, Fernandez-Lahore Marcelo
Downstream Processing Laboratory, School of Engineering and Science, Jacobs University Bremen gGmbH, Campus Ring 1, D-28759 Bremen, Germany.
J Chromatogr A. 2007 Aug 17;1161(1-2):41-50. doi: 10.1016/j.chroma.2007.02.036. Epub 2007 Feb 16.
Protein separation during ion-exchange chromatography implies complex physicochemical events. This work has evaluated the chromatographic behaviour of a complex cell proteome on commercial agarose-based adsorbents. Various ligand types in the cation- and anion-exchange mode were studied. ANX-Sepharose, a weak anion exchanger, performed similarly to the strong anion exchanger-type materials. Proteomic tools were applied in order to understand protein separation. Experimental evidence showed a correlation between apparent isoelectric point distributions and the mobile phase conductivity. Molecular weight distributions were unaffected by the elution position. On the basis of two-dimensional electrophoresis, operational windows were described having typical minor contaminants. These could be annotated for future implementation of in silico downstream processing.
离子交换色谱法中的蛋白质分离涉及复杂的物理化学过程。这项工作评估了复杂细胞蛋白质组在基于琼脂糖的商业吸附剂上的色谱行为。研究了阳离子和阴离子交换模式下的各种配体类型。弱阴离子交换剂ANX-Sepharose的性能与强阴离子交换剂类型的材料相似。应用蛋白质组学工具来理解蛋白质分离。实验证据表明表观等电点分布与流动相电导率之间存在相关性。分子量分布不受洗脱位置的影响。基于二维电泳，描述了具有典型微量污染物的操作窗口。这些可为未来计算机辅助下游处理的实施提供注释。
