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Study on immunocapture–chemiluminescence assay of lipase activity in a biological sample
Author(s) -
Ichibangase Tomoko,
Hamabe Chie,
Ohba Yoshihito,
Kishikawa Naoya,
Nakashima Kenichiro,
Kayamori Yuzo,
Kang Dongchon,
Hamasaki Naotaka,
Kuroda Naotaka
Publication year - 2006
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/bio.885
Subject(s) - lipase , chemiluminescence , horseradish peroxidase , chemistry , chromatography , hydrogen peroxide , triacylglycerol lipase , substrate (aquarium) , luminol , lauric acid , hydrolysis , biochemistry , enzyme , biology , fatty acid , ecology
A new approach for the determination of lipase (triacylglycerol lipase, EC.3.1.1.3) activity in a biological sample was investigated by combining an immunocapture technique with a chemiluminescence (CL) assay method in order to eliminate interference with CL detection. The proposed method consists of an immunocapture step to trap lipase and a subsequent step for CL detection of the activity of the captured lipase. The CL detection is based on the luminol–hydrogen peroxide (H 2 O 2 )–horseradish peroxidase (HRP) reaction and utilizes a proenhancer substrate [a lauric acid ester of 2‐(4‐hydroxyphenyl)‐4,5‐diphenylimidazole (HDI)] which liberates an active enhancer, HDI, by enzymatic hydrolysis. A polyclonal antibody prepared with porcine pancreas lipase was used for the immunocapture. The proposed immunocapture–CL method effectively eliminated the interference with the CL reaction from biological components and enabled the determination of spiked porcine pancreas lipase activity in serum samples in the range 0.41–1.1 U HDI (1 U HDI corresponds to the amount which liberates 1 pmol HDI/min at 37°C from the substrate). The method was further applied to the assay of the activity for human pancreas lipase in serum and the results showed good correlation ( r = 0.871) with those by the conventional colorimetric method. Copyright © 2005 John Wiley & Sons, Ltd.