A novel microscale preparative gel electrophoresis system.

Abstract

Currently, the separation targets of preparative electrophoresis range from milligrams to micrograms of proteins. However, most commercially available preparative electrophoretic instruments function at the milligram level. Although some preparative electrophoretic apparatuses operated at the microgram level, the fractionation results are often unsatisfying because they suffer from low resolution, poor recovery, or a long fractionation time. To address these issues, we developed a novel microscale preparative electrophoresis system that consists of a separation apparatus and an elution apparatus. Protein samples are first loaded onto the separation apparatus and separated over the gel according to the molecular weight of each protein. Then the separation gel is transferred to the elution apparatus and the proteins on the gel are eluted through the thickness of the gel. This system offers the following advantages: (1) high resolution: almost no overlap between the adjacent fractions; (2) a short recovery time: fractionation was performed in 2 hours including separation in 100 min and elution in 20 min and (3) high recovery: recovery was as high as 91.8%.