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Journal of Lipid Research, Vol. 49, 1431-1437, July 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

A novel method for measuring human lipoprotein lipase and hepatic lipase activities in postheparin plasma^*

S. Imamura^*, J. Kobayashi^1,, K. Nakajima^***, S. Sakasegawa^*, A. Nohara, T. Noguchi, M. A. Kawashiri, A. Inazu^**, S. S. Deeb, H. Mabuchi and J. D. Brunzell

^* Diagnostics Research & Development Department, Diagnostic Division, Asahi Kasei Pharma Corporation, Izunokuni City, Japan
Department of Lipidology, Kanazawa University Graduate School of Medical Science, Kanazawa City, Japan
Division of Cardiology, Kanazawa University Graduate School of Medical Science, Kanazawa City, Japan
^** School of Health Science, Laboratory Sciences, Faculty of Medicine, Kanazawa University, Kanazawa City, Japan
Departments of Medicine and Genome Sciences, Division of Medical Genetics, University of Washington, Diagnostic Division, Seattle, WA
Northwest Lipid Metabolism and Diabetes Research Laboratory, Department of Medicine, Division of Metabolism, Endocrinology and Nutrition, University of Washington; Diagnostic Division, Seattle, WA
^*** Otsuka Pharmaceutical Co., Ltd., Tokyo, Japan

^* This study was supported in part by National Institutes of Health Grants HL-64332 and HL30086. Part of this study was performed at the General Clinical Research Center of the University of Washington (National Institutes of Health Grant MO-1 RR37).

Published, JLR Papers in Press, March 14, 2008.

¹ To whom correspondence should be addressed. e-mail: junji{at}med.kanazawa-u.ac.jp

The objective of this study was to establish a new lipoprotein lipase (LPL) and hepatic lipase (HL) activity assay method. Seventy normal volunteers were recruited. Lipase activities were assayed by measuring the increase in absorbance at 546 nm due to the quinoneine dye. Reaction mixture-1 (R-1) contained dioleoylglycerol solubilized with lauryldimethylaminobetaine, monoacylglycerol-specific lipase, glycerolkinase, glycerol-3-phosphate oxidase, peroxidase, ascorbic acid oxidase, and apolipoprotein C-II (apoC-II). R-2 contained Tris-HCl (pH 8.7) and 4-aminoantipyrine. Automated assay of lipase activities was performed with an automatic clinical analyzer. In the assay for HL + LPL activity, 160 µl R-1 was incubated at 37°C with 2 µl of sample for 5 min, and 80 µl R-2 was added. HL activities were measured under the same conditions without apoC-II. HL and LPL activities were also measured by the conventional isotope method and for HL mass by ELISA. Lipase activity detected in a 1.6 M NaCl-eluted fraction from a heparin-Sepharose column was enhanced by adding purified apoC-II in a dose-dependent manner, whereas that eluted by 0.8 M NaCl was not. Postheparin plasma-LPL and HL activities measured in the present automated method had high correlations with those measured by conventional activity and mass methods. This automated assay method for LPL and HL activities is simple and reliable and can be applied to an automatic clinical analyzer.

Supplementary key words dioleylglycerol • quinonediimine dye • automatic clinical analyzer

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