Electrophoretically mediated microanalysis of leucine aminopeptidase using two-photon excited fluorescence detection on a microchip.
Two-photon excited fluorescence detection was performed on a microfabricated electrophoresis chip. A calibration curve of the fluorescent tag beta-naphthylamine was performed, resulting in a sensitivity of 2.5 x 10(9) counts M(-1) corresponding to a detection limit of 60 nM. Additionally, leucine aminopeptidase was assayed on the chip using electrophoretically mediated microanalysis. The differential electroosmotic mobilities of the enzyme and substrate, L-leucine beta-naphthylamide, allowed for efficient mixing in an open channel, resulting in the detection of a 30 nM enzyme solution under constant potential. A zero potential incubation for 1 min yielded a calculated detection limit of 4 nM enzyme.
Zugel
S A
SA
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907-1393, USA.
Burke
B J
BJ
Regnier
F E
FE
Lytle
F E
FE
D000818:Animals; D002138:Calibration; D004586:Electrophoresis / Q000379:methods*; D007931:Leucyl Aminopeptidase / Q000032:analysis*; D008904:Miniaturization; D017785:Photons; D012666:Semiconductors; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013552:Swine
D007931:Leucyl Aminopeptidase