Microchip-based capillary electrophoresis for determination of lactate dehydrogenase isoenzymes.
This article describes a novel microchip-based capillary electrophoresis and oncolumn enzymatic reaction analysis protocol for lactate dehydrogenase (LDH) isoenzymes with a home-made xenon lamp-induced fluorescence detection system. A microchip integrated with a temperature-control unit is designed and fabricated for low-temperature electrophoretic separation of LDH isoenzymes, optimal enzyme reaction temperature control, and product detection. A four-step operation and temperature control are employed for the determination of LDH activity by on-chip monitoring of the amount of incubation product of NADH during the fixed incubation period and at a fixed temperature. Experiments on the determination of LDH standard sample and serum LDH isoenzymes from a healthy adult donor are carried out. The results are comparable with those obtained by conventional CE. Shorter analysis times and a more stable and lower background baseline can be achieved. The efficient separation of different LDH forms indicates the potential of microfluidic devices for isoenzyme assay.
Zhuang
Gui-Sheng
GS
State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, China. guishengzhuang@gmail.com
Liu
Jing
J
Jia
Chun-Ping
CP
Jin
Qing-Hui
QH
Zhao
Jian-Long
JL
Wang
Hui-min
HM
D000328:Adult; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007527:Isoenzymes / Q000097:blood / Q000302:isolation & purification; D007770:L-Lactate Dehydrogenase / Q000097:blood* / Q000302:isolation & purification; D056656:Lab-On-A-Chip Devices; D046208:Microchip Analytical Procedures / Q000379:methods
D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase; C409667:lactate dehydrogenase 1