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J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.D500018-JLR200 on June 16, 2005

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Journal of Lipid Research, Vol. 46, 2015-2022, September 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Methods

A sandwich enzyme-linked immunosorbent assay for human plasma apolipoprotein A-V concentration

Mitsuaki Ishihara*, Takeshi Kujiraoka*, Tadao Iwasaki*, Makoto Nagano*, Mayumi Takano*, Jun Ishii{dagger}, Masahiro Tsuji§, Hajime Ide§, Irina P. Miller**, Norman E. Miller** and Hiroaki Hattori1,*

* Department of Advanced Medical Technology and Development, BML, Inc., Saitama, Japan
{dagger} Division of Internal Medicine, Hokkaido Hospital for Social Health Insurance, Sapporo, Japan
§ Institute of Medical Science, Health Sciences University of Hokkaido, Sapporo, Japan
** Cardiovascular Genetics Division, Department of Internal Medicine, University of Utah School of Medicine, Salt Lake City, UT

Published, JLR Papers in Press, June 16, 2005. DOI 10.1194/jlr.D500018-JLR200

1 To whom correspondence should be addressed. e-mail: hhiro{at}bml.co.jp

Apolipoprotein A-V (apoA-V) is a recently discovered apolipoprotein that appears to have a role in plasma triglyceride (TG) transport. We have developed an ELISA for apoA-V using monoclonal antibodies that has a lower limit of detection of 0.3 ng/ml and linearity up to 20 ng/ml. The ELISA was then used to quantify plasma apoA-V in 196 healthy subjects and 106 patients with insulin-resistant diabetes mellitus. In the healthy subjects, total apoA-V concentration was 179.2 ± 74.8 ng/ml, and it was greater in females than in males (P < 0.005). It was correlated positively with the plasma HDL cholesterol (r = 0.32, P < 0.0001), apoA-I (r = 0.27, P = 0.0001), and apoE (r = 0.18, P = 0.011) concentrations and negatively with plasma TG concentration (r = –0.22, P = 0.021). In relation to single nucleotide polymorphism 3 (–1131C/T) of the apoA-V gene, apoA-V concentration was higher in the T/T type than in the C/C type (P < 0.01). Plasma TG concentration was lower in the T/T type than in the C/C or C/T type (P < 0.05).

ApoA-V concentration was lower in the diabetic patients (69.4 ± 44.3 ng/ml; P < 0.01) than in the healthy controls.

Abbreviations: apoA-V, apolipoprotein A-V; CBB, Coomassie brilliant blue; MAb, monoclonal antibody; NIDDM, non-insulin-dependent diabetes mellitus; rhapoA-V, recombinant human apolipoprotein A-V; SNP, single nucleotide polymorphism; SREBP-1c, sterol-regulatory element binding protein 1c; TG, triglyceride

Supplementary key words triglyceride • diabetes mellitus • immunoassay • single nucleotide polymorphism


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