Large-scale preparative disk gel electrophoresis with electroosmotic medium pump system.

Abstract

We constructed a large-scale preparative gel electrophoresis device equipped with a 10-cm in diameter gel support column and used it with an electroosmotic medium pump system (EOMPS). Operation of the constructed apparatus was examined and resolution of the protein bands was determined by separation of the marker protein mixture (6.4-198kDa) using SDS-PAGE (Laemmli buffer system). In addition, loading capacity was tested by fractionation of 1-2g of a bovine serum albumin (BSA) Cohn V fraction using native-PAGE, resulted in a reasonable separating profile. The advantages of the present apparatus were considered to be its good resolving power combined with simplicity, despite the large scale, and a high loading capacity. An EOMPS enhances the possibility of continuous electrophoretic fractionation of complex protein mixtures on a large scale, and the present apparatus is able to purify nearly all types of protein using SDS-PAGE as well as acidic proteins with native-PAGE.