Analysis of lipoprotein lipase activity using high-performance liquid chromatography.
Lipoprotein lipase (LPL) is a key enzyme which regulates the plasma triglyceride concentration by hydrolyzing triglycerides in chylomicrons and very-low-density lipoprotein (VLDL). The activity of LPL was conventionally analyzed using radio-labeled residues or direct sandwich-ELISA. An assay for lipoprotein lipase activity which used a nonradioactive substrate, tri-olein, is described. In this method, LPL activity was detected fluorometrically by reacting 9-anthryldiazomethane (ADAM) with the oleic acid generated from tri-olein by enzyme activity and separated by reversed-phase HPLC. This method has been optimized and the optimum enzyme incubation time and reaction time of the generated oleic acid with ADAM were both at 20 min. The method correlated well with the conventional method.
Eguchi
Yukinori
Y
Research Laboratory Center, Faculty of Medicine, University of the Ryukyus, Uehara 207, Nishihara-chou, Okinawa, 903-0125, Japan. yeguchi@med.u-ryukyu.ac.jp
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008071:Lipoprotein Lipase / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D008071:Lipoprotein Lipase