Ogle David, Ho Agnes, Gibson Toby, Rylatt Dennis, Shave Evan, Lim Peniel, Vigh Gyula
Gradipore, French's Forest, NSW 2086, Australia.
J Chromatogr A. 2002 Dec 6;979(1-2):155-61. doi: 10.1016/s0021-9673(02)01601-1.
The Gradiflow BF200 preparative electrophoretic unit (Gradipore), which has been developed for size-based and charge-sign-based protein separations and in which the hydraulic flow path of the recirculating sample stream in the separation cartridge is orthogonal to the electric field, has been modified to carry out binary protein separations using the principles of isoelectric trapping. The disposable separation cartridge contained three isoelectric membranes which, along with the cartridge holder, formed the anode and cathode compartments and the anodic and cathodic separation compartments. The utility of the modified instrument was demonstrated by effecting a binary separation of chicken egg white across an isoelectric point 5.5 isoelectric membrane. The desalting and subsequent binary separation steps proved to be remarkably rapid, due to the favorable combination of short electrophoretic path, high electric field strength and large effective isoelectric membrane surface area.
Gradiflow BF200 制备型电泳装置（Gradipore）专为基于大小和电荷信号的蛋白质分离而开发，其分离柱中循环样品流的液压流路与电场正交，现已进行改进，以利用等电捕获原理进行二元蛋白质分离。一次性分离柱包含三个等电膜，这些等电膜与柱架一起形成阳极和阴极隔室以及阳极和阴极分离隔室。通过在等电点为5.5的等电膜上对鸡蛋白进行二元分离，证明了改进后仪器的实用性。由于电泳路径短、电场强度高和有效等电膜表面积大等有利因素的结合，脱盐及随后的二元分离步骤被证明非常迅速。
