Electrophoretically mediated enzyme analysis in capillaries
Abstract
Electrophoretically mediated microanalysis for enzymes was developed in silica capillaries. The separation power of CZE (Capillary zone electrophoresis) gave advantages to EMMA (Electrophoretically mediated micro-analysis) over spectroscopic methods for crude samples such as Sigma enzyme control containing hundreds of other proteins and urine samples containing significant amounts of chromogens which interfere with the target analyte. Very small amounts of enzyme and substrate (less than 1 $\mu{\rm l}$ per run) were mixed electrophoretically in the capillary to generate enzyme reaction products. Since only the effective separation length of the capillary is required to be filled with enzyme, this system uses a tiny amount of enzyme. A shorter effective length and narrower i.d. capillary will save more material. The method shows an interesting feature in that one can move the target peak very easily to obtain optimum noise-free data by controlling the running time before the incubation. The theoretical model and equations were derived for the selectivity in EMMA with the leucine aminopeptidase enzyme system.
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Analytical chemistry|Biochemistry
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