Application of electrophoretically mediated microanalysis for enzyme assays

Application of capillary electrophoresis for the study of enzymes Mgr. Soňa Vytisková, Ph.D Enzymes are biological catalysts that play an important role in biochemical reactions necessary for normal growth, maturation and reproduction through the whole living world. They catalyze virtually all chemical reactions in living systems and the assay of enzyme activity is probably one of the most frequently encountered procedures in biochemistry and molecular biology. This work aimed to develop capillary electrophoresis (CE)-based assay for different enzyme, thereby exploring the capabilities of CE-UV regarding this application. The enzymatic systems included in this project are all clinically relevant but differential with respect to the type and number of substrates, products and cofactors. In order to investigate these enzymes, model substrates that are converted into UV detectable products were used. CE is a powerful and relatively new analytical tool. The small dimensions of the CE separation systems utilized are a primary advantage for the bio-research. A recent trend is the use of CE in the area of immunoassays and enzyme assays, due to its fast analysis time and the minute amounts of sample. Especially in-capillary approaches to study of enzyme activity, such as electrophoretically mediated..