Study on immunocapture–chemiluminescence assay of lipase activity in a biological sample

✍ Scribed by Tomoko Ichibangase; Chie Hamabe; Yoshihito Ohba; Naoya Kishikawa; Kenichiro Nakashima; Yuzo Kayamori; Dongchon Kang; Naotaka Hamasaki; Naotaka Kuroda

• Publisher: John Wiley and Sons
• Year: 2006
• Tongue: English
• Weight: 114 KB
• Volume: 21
• Category: Article
• ISSN: 1522-7235
• DOI: 10.1002/bio.885

✦ Synopsis

Abstract

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.