Centrifugal Precipitation Chromatography

介绍了一种新的色谱系统。的 系统内部产生铵的浓度梯度 硫酸盐（AS）沿着长通道使蛋白质断裂， 其在AS溶液中的溶解度。分离柱包括： 一对具有相互镜像的螺旋通道的盘， 被半透膜隔开。所述盘组件 安装在无密封连续流动离心机上。浓缩砷 溶液被引入上通道， 在相反的方向上通过下通道， 旋转柱将蛋白质混合物注入水中 通道沿着浓度增加的AS梯度移动 已经在水溶液中建立。每种蛋白质 沿梯度沿着以不同的AS浓度沉淀。的 使用馏分连续监测和收集 收藏家该方法已被证明在分离 血清蛋白并应用于纯化重组 从E.大肠杆菌裂解物， 亲和配体的样品溶液。

A novel chromatographic system is introduced. The system internally generates a concentration gradient of ammonium sulfate (AS) along a long channel to fractionate proteins according to their solubility in AS solution. The separation column consists of a pair of disks with mutually mirror-imaged spiral channels that are separated by a semipermeable membrane. The disk assembly is mounted on a sealless continuous flow centrifuge. Concentrated As solution is introduced into the upper channel while a water solution is passed through the lower channel in the opposite direction in a rotating column. A mixture of proteins injected into the water channel moves along an AS gradient of increasing concentration that has been established in the water solution. Each protein species precipitates at a different AS concentration along the gradient. The eluate is continuously monitored and collected using a fraction collector. The method has been demonstrated on separation of serum proteins and applied to purification of a recombinant ketosteroid isomerase from a crude E. coli lysate by adding an affinity ligand to the sample solution.