Membrane electrophoresis

Abstract

Abstract The separation of substances by membrane electrophoresis using the principle of isoelectric focussing is improved by the use of membranes of known isoelectric point and having buffering capacity. When such membranes are arranged in series of increasing isoelectric point from the anode (+) to the cathode (-) in a pH gradient of increasing pH in the same direction, and each membrane is located between electrolyte zones having pHs respectively lower and higher than its isoelectric point, the tendency for electro-endosmosis to occur during electrophoresis is substantially reduced. Methods for chemical treatment of agarose and other gel membranes to introduce acidic and basic groups so as to confer buffering power at the isoelectric point are described. These include the attachment of ampholytes to the gel by intermediate chemical linkages. Under varying chemical conditions membranes exhibiting isoelectric properties over a range of pH values can be prepared and specific values predetermined to within narrow limits. The membranes are used in multicompartment apparatus with separate cooling of individual circulating electrolyte zones and may be in the form of sheet membranes mounted in a nesting configuration one above the next with gravity flow of electrolyte and edge sealing between the membranes. High current and large scale use of the method for preparative as well as analytical purposes are envisaged.

Inventors

• 👤 Archer J. P. Martin
• 👤 Frank Hampson

Bibliographic Details