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Animal Bioresource in Japan

.: Home > Animal Bioresource in Japan > 2010 > Volume 59 Number 4 > Makoto KINOSHITA1), Teruhiko MATSUSHIMA2), Yamato MASHIMO1), Masaaki KOJIMA3), Megumi KIGURE3) and Tamio TERAMOTO1)

Determination of Immuno-Reactive Rabbit Apolipoprotein B-48 in Serum by ELISA

Makoto KINOSHITA1), Teruhiko MATSUSHIMA2), Yamato MASHIMO1), Masaaki KOJIMA3), Megumi KIGURE3) and Tamio TERAMOTO1)
1) Department of Internal Medicine, Teikyo University School of Medicine 2) Internal Medicine, Tsukuba Memorial Hospital 3) Shibayagi Co., Ltd.
Abstract :

Apolipoprotein B-48 (apoB-48) is produced by the small intestine, is associated with chylomicrons, and appears to be a suitable marker for clinical studies of postprandial dynamics of lipoproteins. It is also associated with cardiovascular risk factors. We have developed an assay system to quantify immuno-reactive apoB-48 in rabbit serum or plasma. A microtiter-plate was coated with monoclonal antibody raised against human apoB-48 C-terminal specific decapeptide that has high homology to the rabbit C-terminal sequence. Appropriate ELISA standard curves were obtained using apoB-48 extracted from rabbit serum by immuno-affinity chromatography. No cross-reactivity was found with apoB-100 in western blot analyses. Intra- and inter-assay CVs were less than 3%. Recovery of rabbit apoB-48 spiked in serum was within 93.4–105%. ApoB-48 levels in healthy controls rabbits fed a normal diet were within the range of 0.903–1.09 μg/ml (mean ± SD: 1.03 ± 0.084 μg/ml). In healthy animals, the blood apoB-48 level was markedly increased by a high fat diet and in the postprandial state in parallel with serum triglyceride. Ezetimibe, cholesterol absorption inhibitor, given orally to rabbits fed on a high fat diet blocked further increase of blood levels of apoB-48 and triglyceride. This method for measuring apoB-48 using the monoclonal antibody is simple, reliable, and suitable for routine analyses.

Keywords :
ELISA, ezetimibe, monoclonal-antibody, rabbit apolipoprotein B-48

Date Deposited : 21 Jun 2011 14:33

Last Modified : 21 Jun 2011 14:33

Official URL: http://www.jstage.jst.go.jp/browse/expanim/60/1/_contents

Volume 59, Number 4, - 2010

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