The University of Texas System

Ampholife Technologies

G David Huff

L Scott Rodkey

The present invention includes novel isoelectric focusing cells and methods for isoelectric focusing of amphoteric substances within fluids containing carrier ampholytes which inhibit the phenomena known as spiking or acid notching. The apparatus includes a barrier separating the electrode compartments from the focusing cell compartments which minimizes hydrogen and hydroxyl ions contained in electrolyte solution within the electrode chambers from passing into the focusing cell passageways. The currently preferred barrier is a bipolar membrane; however, the combination of anion and cation selective membranes is also effective. Such a barrier enables very narrow pH gradients to be established and maintained, thereby permitting high resolution separations of amphoteric biological substances.

Apparatus and methods for isoelectric focusing of amphoteric substances

The present invention includes novel apparatus and methods for isoelectric focusing of amphoteric substances within fluids containing carrier ampholytes. The present invention utilizes multichannel recycling isoelectric focusing techniques which eliminate mixing of fluid which passes through the isoelectric focusing cell with the original crude fluid sample. Mixing is avoided by preferably using a dual reservoir system and alternating the fluid flow into and out of each reservoir after each pass through the isoelectric focusing cell. Fluid flow is alternated into and out of each reservoir of each reservoir pair through use of reservoir inlet and outlet valves. The reservoir inlet and outlet valves preferably operate in response to the fluid level within the reservoir supplying fluid to the isoelectric focusing cell. When the reservoir empties the corresponding reservoir outlet valve automatically engages to stop fluid flow from the empty reservoir and to start fluid flow from the full reservoir. Near instantaneously therewith, the reservoir inlet valve engages to stop fluid flow into the full reservoir and to start fluid flow into the empty reservoir. The isoelectric focusing cell preferably has a plurality of inlet ports and a plurality of corresponding outlet ports. The inlet and outlet ports permit fluid to flow into and out of the isoelectric focusing cell along discrete channels. Each inlet and outlet port is then coupled to a pair of the alternating reservoirs. The apparatus within the scope of the present invention preferably includes a heat exchanger for removing heat generated by the isoelectric focusing cell. The heat exchanger is preferably coupled directly to the isoelectric focusing cell thereby permitting greater power input to the focusing cell which results in rapid focusing of both the carrier ampholytes and the fluid sample.

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Houston, TX, United States of America

G David Huff