Multi-compound electrophoretic assays for tyramine oxidase with a UV area detector imaging multiple windows on a looped capillary.
Active pixel sensor UV area imaging and capacitively coupled contactless conductivity detection have been applied in an electrophoretically mediated microanalysis (EMMA) assay for substrate specificity of tyramine oxidase (Arthrobacter sp.). Use of the UV area imaging detector to monitor four windows in a capillary with three loops provided intrinsic self-referencing for all species and identified tyramine and 2-phenethylamine as the only reactive components in a multi-compound mixture. Continuous engagement EMMA experiments showed significant benefits by comparison with plug-plug EMMA, improving sensitivity by extending enzyme-substrate interaction times and allowing measurement of time-dependent reaction in the substrate zones passing the four windows.
Urban
Pawel L
PL
Department of Chemistry, University of York, Heslington, York YO10 5DD, UK.
Goodall
David M
DM
Carvalho
Alexandre Z
AZ
Bergström
Edmund T
ET
Van Schepdael
Ann
A
Bruce
Neil C
NC
D001173:Arthrobacter / Q000201:enzymology; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D008995:Monoamine Oxidase / Q000378:metabolism*; D010627:Phenethylamines; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation / Q000379:methods*; D013379:Substrate Specificity; D014439:Tyramine / Q000378:metabolism
D010627:Phenethylamines; C029261:phenethylamine; D008995:Monoamine Oxidase; D014439:Tyramine