Study on immunocapture-chemiluminescence assay of lipase activity in a biological sample.
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 degrees 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.
Ichibangase
Tomoko
T
Graduate School of Biomedical Sciences, Course of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Hamabe
Chie
C
Ohba
Yoshihito
Y
Kishikawa
Naoya
N
Nakashima
Kenichiro
K
Kayamori
Yuzo
Y
Kang
Dongchon
D
Hamasaki
Naotaka
N
Kuroda
Naotaka
N
D003124:Colorimetry; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D008049:Lipase / Q000097:blood / Q000378:metabolism*; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry; D010179:Pancreas / Q000201:enzymology; D012680:Sensitivity and Specificity
D008165:Luminol; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D008049:Lipase