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A more recent version of this article appeared on July 1, 2008

Papers In Press, published online ahead of print March 14, 2008
J. Lipid Res., doi:10.1194/jlr.M700528-JLR200
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Submitted on November 16, 2007
Revised on February 22, 2008
Accepted on March 14, 2008

A novel method for measuring human lipoprotein lipase and hepatic lipase activities in post-heparin plasma

Shigeyuki Imamura, Junji Kobayashi, Katsuyuki Nakajima, Shinichi Sakasegawa, Atsushi Nohara, Tohru Noguci, Masa-aki Kawashiri, Akihiro Inazu, Samir Deeb, Hiroshi Mabuchi, and John D. Brunzell

Department of lipidology, Kanazawa University Graduate School of Medical Science, Kanazawa city, Ishikawa 920-8640

Corresponding Author: 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 546nm 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. R-2 contained Tris-HCl (pH8.7) and 4-aminoantipyrine. Automated assay of lipase activities was performed with automatic clinical analyzer. In the assay for HL+LPL activity, 160 µl R-1 was incubated at 37  with 2 µl of samples for 5 min, and 80 µl R-2 was added. HL activities were measured in the same condition without apolipoprotein C-II. HL and LPL activities were also measured by the conventional isotope method and for HL mass by ELISA. Lipase activity detected in 1.6 M NaCl eluted fraction from a heparin-Sepharose was enhanced by adding purified apolipoprotein C-II in a dose dependent manner, whereas that eluted by 0.8 M NaCl was not. PHP-LPL and HL activities measured in the present automated method had high correlations with those measured by the conventional activity and mass methods. This automated assay method for LPL and HL activities is simple and reliable and can be applied to automatic clinical analyzer.


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