Electrophoretically mediated microanalysis

Lee, I.H., Pinto, D., Arriaga, E.A., Zhang. Z., Dovichi, NJ. (1998). Picomolar analysis of proteins using electrophoretically mediated microanalysis and capillary electrophoresis with laser-induced fluorescence detection. Anal. Chem. 70, 4546 4548. 

ABEI, M(4-ami nobutyl )-Methylisolu mi nol BSA, bovine serum albumin CL, chemiluminescence DNPO, tas-(2,4-dinitrophenyl)oxalate ECL, electrogenerated chemiluminescence EMMA, electrophoretically mediated microanalysis EY, eosine Y FR, lluorescamine HRP, horseradish peroxidase ILITC, isoluminol isothiocyanate LOD, limit of detection RITC, rhodamine B isothiocyanate TCPO, Mv-(2,4,6-trichlorophenyl)oxalate TEA, triethylamine TRITC, tetramethylrhodamine isothiocyanate. 

The same reaction was recently proposed to detect creatine kinase (CK), an enzyme of high clinical significance in relation to the investigation of skeletal muscle disease and the diagnosis of myocardial infarct or cerebrovascular accidents. As ATP is a reaction product obtained from the reaction of ADP with creatine phosphate catalyzed by CK, this enzyme can be indirectly measured by the CL intensity read from the subsequent reaction of ATP with luciferin. Using the technique of electrophoretically mediated microanalysis (EMMA), it is possible to detect the enzyme using nanoliter volumes of biological sample with an improved speed and simplicity with respect to a conventional colorimetric method [100],... 

Electrogenerated chemiluminescence Enzyme-linked immunosorbent assay Electrophoretically-mediated microanalysis Electroosmotic flow Eosine Y... 

Kaale, E., Van Goidsenhoven, E., Van Schepdael, A., Roets, E., and Hoogmartens, J. (2001). Electrophoretically mediated microanalysis of gentamicin with in-capillary derivatization and UV detection. Electrophoresis 22, 2746—2754. 

Kiessig et al. studied the interaction of the immunosuppressant drug cyclosporin A and some derivatives to its cellular receptor protein cyclo-philin (26). The cyclophilin-cyclosporin A complex mediates the immunosuppressive action of cyclosporin A by inhibition of the phosphatase calci-neurin. Using ACE in the equilirium-mixture mode, a separation of the cyclophilin-cyclosporin A complex and the unbound cyclophilin was achieved. The approach allowed a qualitative estimation of the binding affinity of cyclosporin A derivatives compared to cyclosporin A. For the calculation of binding constants, electrophoretically mediated microanalysis was applied. 

Zugel, S.A., Burke, B.J., Regnier, F.E., Lytle, F.E., Electrophoretically mediated microanalysis of leucine aminopeptidase using two-photon excited fluorescence detection on a microchip. Anal. Chem. 2000, 72, 5731-5735. 

Most applications of native fluorescence detection involve the direct detection of a fluorescent analyte of interest. However, native fluorescence can also be used indirectly to detect the presence of nonfluorescent species. For example, the presence of a nonfluorescent enzyme can be detected based on its conversion of substrate into natively fluorescent products. This is often used in conjunction with electrophoretically mediated microanalysis, in which differences in buffer mobilities enable on-column mixing and reaction of enzymes with substrates. One such study exploited the intrinsic fluorescence of nicotinamide adenine dinucleotide (NADH) to investigate differences in the reactivity of individual molecules of lactate dehydrogenase. 

Vallejo-Cordoba, B., Mazorra-Manzano, M. A., and Gonzalez-Cordova, A. R, Determination of P-hydroxyacyl CoA-dehydrogenase activity in meat by electrophoretically mediated microanalysis, J. Cap. Elec. Micro. Tech., 8, 81, 2003. 

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