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Titolo:
Fluorogenic assay for beta-glucuronidase using microchip-based capillary electrophoresis
Autore:
Starkey, DE; Han, A; Bao, JJ; Ahn, CH; Wehmeyer, KR; Prenger, MC; Halsall, HB; Heineman, WR;
Indirizzi:
Procter & Gamble Pharmaceut, Hlth Care Res Ctr, Mason, OH 45040 USA Procter & Gamble Pharmaceut Mason OH USA 45040 s Ctr, Mason, OH 45040 USA Univ Cincinnati, Dept Elect & Comp Engn & Comp Sci, Cincinnati, OH 45221 USA Univ Cincinnati Cincinnati OH USA 45221 omp Sci, Cincinnati, OH 45221 USA Univ Cincinnati, Dept Chem, Cincinnati, OH 45221 USA Univ Cincinnati Cincinnati OH USA 45221 pt Chem, Cincinnati, OH 45221 USA
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 1, volume: 762, anno: 2001,
pagine: 33 - 41
SICI:
1387-2273(20011005)762:1<33:FAFBUM>2.0.ZU;2-P
Fonte:
ISI
Lingua:
ENG
Soggetto:
FLUORESCEIN MONOGLUCURONIDE; SAMPLE INJECTION; PCR PRODUCTS; GLASS CHIPS; HUMAN-URINE; SYSTEMS; PERFORMANCE; IMMUNOASSAY; QUANTIFICATION; THEOPHYLLINE;
Keywords:
microchips; CE; beta-glucuronidase; enzymes;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Wehmeyer, KR Procter & Gamble Pharmaceut, Hlth Care Res Ctr, 8700 Mason Montgomery Rd, Mason, OH 45040 USA Procter & Gamble Pharmaceut 8700 Mason Montgomery Rd Mason OH USA 45040
Citazione:
D.E. Starkey et al., "Fluorogenic assay for beta-glucuronidase using microchip-based capillary electrophoresis", J CHROMAT B, 762(1), 2001, pp. 33-41

Abstract

Microchip capillary electrophoresis (CE) was used with a model enzyme assay to demonstrate its potential application to combinatorial drug screening. Hydrolysis with beta -glucuronidase of the conjugated glucuronide, fluorescein mono-beta -D-glucuronide (FMG), liberated the fluorescent product, fluorescein. FMG and fluorescein were detected by fluorescence, with excitation and emission at 480 and 520 nm, respectively. Microchip CE was used to separate FMG and fluorescein. Fluorescein production was monitored to assess beta -glucuronidase activity. Michaelis-Menten enzyme kinetics analysis yielded the K-m value. The results were compared with those from experiments done by conventional CE. The K-m value for beta -glucuronidase with FMG is being reported for the first time as 18 muM. The inhibition of beta -glucuronidase by the competitive inhibitor D-saccharic acid-1,4-lactone (SL) was also determined using microchip CE. Reactions were done with various concentrations of inhibitor and constant beta -glucuronidase and FMG concentrations. A dose-response plot was acquired and the IC50 value for SL was determined to be 3 muM. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 22/01/20 alle ore 13:25:23