Electrophoretically mediated microanalysis of a nicotinamide adenine dinucleotide-dependent enzyme and its facile multiplexing using an active pixel sensor UV detector

被引:15
作者
Urban, Pawel L.
Goodall, David M. [1 ]
Bergstroem, Edmund T.
Bruce, Neil C.
机构
[1] Univ York, Dept Chem, York YO10 5DD, N Yorkshire, England
[2] Paraytec Ltd, York YO04 1GN, N Yorkshire, England
[3] Univ York, Dept Biol, CNAP, York YO10 5DD, N Yorkshire, England
关键词
active pixel sensor; capillary electrophoresis; EMMA; enzyme assay; NAD; plug-plug; CAPILLARY-ZONE-ELECTROPHORESIS; ASSAY; COMBINATORIAL; WAVELENGTH; ABSORBENCY; EVOLUTION; CE;
D O I
10.1016/j.chroma.2007.04.071
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An electrophoretically mediated microanalysis (EMMA) method has been developed for yeast alcohol dehydrogenase and quantification of reactant and product cofactors, NAD and NADH. The enzyme substrate ethanol (1% (v/v)) was added to the buffer (50 mM borate, pH 8.8). Results are presented for parallel capillary electrophoresis with a novel miniature UV area detector, with an active pixel sensor imaging an array of two or six parallel capillaries connected via a manifold to a single output capillary in a commercial CE instrument, allowing conversions with five different yeast alcohol dehydrogenase concentrations to be quantified in a single experiment. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:132 / 140
页数:9
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