Dynamic modeling of electrophoretically mediated microanalysis

被引:46
作者
Patterson, DH [1 ]
Harmon, BJ [1 ]
Regnier, FE [1 ]
机构
[1] PURDUE UNIV,DEPT CHEM,W LAFAYETTE,IN 47907
关键词
electrophoretically mediated microanalysis; mathematical modelling; electromigration; band broadening; computer simulation; diffusion; reaction kinetics; ethanol; leucine aminopeptidase; aminopeptidases; enzymes;
D O I
10.1016/0021-9673(95)01247-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A dynamic model is presented for simulation of reaction-based chemical analysis of enzymes and substrates in capillary electrophoretic systems by the methodology of electrophoretically mediated microanalysis (EMMA). The mathematical model utilizes mass balance expressions describing the time-dependent effects of electromigration, chemical reaction, and diffusional band broadening upon the concentration profiles of the various reagent and product species. The model is implemented in an iterative computer program in which the capillary is segmented into arrays of bins storing the concentration profiles of each of the chemical species. During each time increment, the effects of electrophoresis, reaction kinetics, and diffusion are calculated, and the concentrations stored in the arrays are updated. The flexibility of the model to accommodate various initial capillary conditions, sample introduction methods, and voltage programming allows diverse EMMA analyses to be simulated. The simulated results are shown to be in good qualitative agreement with experimental data for zonal injection and moving boundary EMMA determinations of leucine aminopeptidase as well as an EMMA analysis of ethanol.
引用
收藏
页码:119 / 132
页数:14
相关论文
共 26 条
[21]   ANALYTICAL AND MICROPREPARATIVE SEPARATION OF PEPTIDES BY CAPILLARY ZONE ELECTROPHORESIS USING DISCONTINUOUS BUFFER SYSTEMS [J].
SCHWER, C ;
LOTTSPEICH, F .
JOURNAL OF CHROMATOGRAPHY, 1992, 623 (02) :345-355
[22]  
WALBROEHL Y, 1989, J MICROCOLUMN SEP, V1, P41
[23]   PRODUCT INHIBITION STUDIES ON YEAST AND LIVER ALCOHOL DEHYDROGENASES [J].
WRATTEN, CC ;
CLELAND, WW .
BIOCHEMISTRY, 1963, 2 (05) :935-&
[24]   ELECTROPHORETICALLY MEDIATED MICROASSAY OF ALKALINE-PHOSPHATASE USING ELECTROCHEMICAL AND SPECTROPHOTOMETRIC DETECTION IN CAPILLARY ELECTROPHORESIS [J].
WU, D ;
REGNIER, FE ;
LINHARES, MC .
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS, 1994, 657 (02) :357-363
[25]   NATIVE PROTEIN SEPARATIONS AND ENZYME MICROASSAYS BY CAPILLARY ZONE AND GEL-ELECTROPHORESIS [J].
WU, D ;
REGNIER, FE .
ANALYTICAL CHEMISTRY, 1993, 65 (15) :2029-2035
[26]   VARIABILITY OF INTRACELLULAR LACTATE-DEHYDROGENASE ISOENZYMES IN SINGLE HUMAN ERYTHROCYTES [J].
XUE, QF ;
YEUNG, ES .
ANALYTICAL CHEMISTRY, 1994, 66 (07) :1175-1178