Centrifugal Precipitation Chromatography

设计了一种新型离心沉淀色谱分离柱，并对人血清白蛋白和丙种球蛋白进行了分离试验。分离柱由市售中空纤维透析装置（MicroCross，阿默舍姆）制成。一对这样的透析筒以螺旋方式固定地安装在旋转板上，使得硫酸铵浓度梯度在离心力场下稳定地保持。 每个透析盒有4个开口：两个开口连接到中空透析纤维作为硫酸铵通道，另外两个开口连接到透析中空纤维的外部，用于洗脱磷酸盐缓冲液和样品蛋白。开始时，使用一个钉匣装置测试分离，但后来将两个装置连接并用于分离。发现该系统的性能与使用透析膜的先前系统完全不同，因为中空纤维透析管更自由地通过水，使得样品倾向于被稀释。为了防止样品的过度稀释，硫酸铵通道的流管使用较大孔径的管。 使用上述设备的实验目前正在我们的实验室进行。

A new separation column for centrifugal precipitation chromatography was designed and tested for separation of human serum albumin and gamma-globulin. The separation column was made of commercially-available hollow-fiber dialysis devices (MicroCross, AmerSham). A pair of these dialysis cartridges are mounted symmetricaly on the rotary plate in a spiral fashion so that the ammonium sulfate concentration gradient is stably retained under a centrifugal force field. Each dialysis cartridge has 4 opinings: two openings connected to hollow dialysis fiber were used as an ammonium sulfate channel and other two openings connected to outside of dialysis hollow fiber for eluting phosphate buffer and sample proteins. At begining One cartridge device was used for testing the separation, but later both devices were connected and used for separation. It was found that the performance of the system is quite different from the previous system using dialysis membrane, since the hollow fiber dialysis tubing passes water more freely so that the samples tend to be diluted. In order to prevent excessive dilution of the samples, the larger bore tubing were used fro flow tubes for the ammonium sulfate channel. The experiment using the above device is currently underway in our laboratory.