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

Tomoko Ichibangase, Chie Hamabe, Yoshihito Ohba, Naoya Kishikawa, Kenichiro Nakashima, Yuzo Kayamori, Dongchon Kang, Naotaka Hamasaki, Naotaka Kuroda

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

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 (H2O2)-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 UHDI 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.

Original languageEnglish
Pages (from-to)62-66
Number of pages5
JournalLuminescence
Volume21
Issue number1
DOIs
Publication statusPublished - Jan 1 2006

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Chemistry (miscellaneous)

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