Centrifugal Precipitation Chromatography

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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.