High performance multidimensional proteome analyzer

Abstract

A proteome analyzer includes a separation cassette having a first dimension separation compartment for separation of protein samples by isoelectric focusing and a second dimension separation compartment for separation of protein samples by SDS-polymer network electrophoresis. The first dimension compartment is a reservoir in which a porous material having capillary channels is disposed. The protein samples are disposed in the capillary channels and, in the presence of a pH gradient, are focused spatially by isolectric focusing upon application of an electric field. The second dimension compartment consists of two glass or plastic plates separated by a separation medium. The separation medium is an ultra-thin layer of a low concentration linear polymer supported by an inert matrix. The spatially focused protein samples are contacted with the separation medium in the presence of an electric field to initiate second dimension separation. The migrating SDS-protein complexes are fluorescently labeled by a dye in the separation medium during second dimension separation, and are detected by an illumination and detection system as they pass through a detection area. The detected SDS-protein complexes are imaged and displayed by computer analysis as two-dimensional maps representing the proteins in the sample.