Capillary zone electrophoretic analysis of isoenzymes

Chemistry: electrical and wave energy – Processes and products – Electrostatic field or electrical discharge

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Details

204299R, G01N 2726, G01N 27447, B01D 5702

Patent

active

052640957

ABSTRACT:
Disclosed herein is a methodology for analyzing isoenzymes using capillary zone electrophoresis ("CZE") techniques. Briefly, an isoenzyme-containing sample and a substrate capable of being catalyzed by said isoenzyme into a reaction product are introduced into a capillary column comprising a buffer. Most preferably, the buffer contains the substrate prior to introduction of the sample into such substrate-buffer. CZE separation techniques are applied to the column such that the isoenzymes are separated from each other into discrete zones. The separation techniques are terminated such that product is rapidly generated by the catalytic conversion of substrate by the isoenzymes, and accumulated, within each discrete zone, followed by detection of product. Information regarding the relative distribution of the isoenzymes can be derived from the relative distribution of the product.

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A. Kopwillem, "Analytical isotachophoresis in capillary tubes: Transformation of pyruvate to succinate by calf heart mitochrondrial enzymes" Journal of Chromatography, 82(1973) 407-409.
Stellan Hjerten and Ming-DeZhu, "Micropreparative Version of High Performance Electrophoresis: The Electrophoretic Counterpart of Narrow-Bore High-Performance Liquid Chromatography" Journal of Chromatography 327 (1985) 157-164.
Peter Stehle, Hans-Peter Bahsitta & Peter Furst, "Analytical Control of Enzyme-Catalyzed Peptide Synthesis Using Capillary Isotachophoresis" Journal of Chromatography 370 (1986) 131-138.
Beckman Product Bulletin 6234, "The Appraise.RTM. and Paragon.RTM. Electrophoresis System".

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