Multiplexed enzyme assays in capillary electrophoretic single-use microfluidic devices.
We describe a method of performing multiple enzyme assays in a single reaction vessel. The resolving power of capillary electrophoresis enables several enzyme assays to be analyzed at high speed in microfluidic arrays. Multiplexed measurement can increase throughput significantly without requiring highly dense microfluidic arrays. Enzyme assays in a multiplexed format for selected kinases in this work show essentially identical performance to assays performed individually. This establishes an approach for screening one compound against multiple enzyme targets simultaneously. Another potential application for performing multiplexed enzyme assay is to study protein-protein (especially enzyme-enzyme) interaction by monitoring the enzymatic activity changes.
Xue
Q
Q
ACLARA BioSciences, Inc, Mountain View, CA, USA. qxue@aclara.com
Wainright
A
A
Gangakhedkar
S
S
Gibbons
I
I
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis; D017868:Cyclic AMP-Dependent Protein Kinases / Q000032:analysis / Q000037:antagonists & inhibitors; D004209:Disposable Equipment; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004791:Enzyme Inhibitors / Q000032:analysis; D004798:Enzymes / Q000032:analysis*; D004867:Equipment Design; D016650:Fluorescein-5-isothiocyanate / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D005470:Fluorometry / Q000295:instrumentation; D047908:Intracellular Signaling Peptides and Proteins; D007700:Kinetics; D007834:Lasers; D008832:Microchemistry / Q000295:instrumentation*; D018613:Microscopy, Confocal / Q000295:instrumentation; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010740:Phospholipases / Q000032:analysis; D011494:Protein Kinases / Q000032:analysis; D012212:Rheology; D019061:src-Family Kinases / Q000032:analysis / Q000037:antagonists & inhibitors
D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D004798:Enzymes; D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins; D010446:Peptide Fragments; C094636:protein kinase inhibitor (6-22); D011494:Protein Kinases; D019061:src-Family Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D010740:Phospholipases; D016650:Fluorescein-5-isothiocyanate