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

doi:10.1194/jlr.D500018-JLR200

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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, Masahiro Tsuji, Hajime Ide, Irina P. Miller, Norman E. Miller, and Hiroaki Hattori

Advanced Medical Technology and Development, BML, Inc., Kawagoe, Saitama 350-1101

Corresponding Author: hhiro{at}bml.co.jp

Apolipoprotein (apo) A-V is a new member of the apolipoprotein gene family and has a role in triglyceride metabolism. We have developed an ELISA for apo A-V, in which the lower limit of detection is 0.3 ng/mL, with linearity up to 20 ng/mL, and have quantified plasma apo A-V concentration in healthy and diabetic subjects. In healthy subjects plasma total apo A-V concentration was 179.2 ± 74.8 ng/mL, and it was higher in females than in males (P<0.005). Plasma apo A-V concentration was correlated positively with high density lipoprotein (HDL) cholesterol (r=0.316, P<0.0001), apo A-I (r=0.269, P=0.0001) and apo E (r=0.180, P=0.011), and negatively with triglycerides (r=-0.218, P=0.021). In females but not males, apo A-V concentration was correlated positively with HDL cholesterol (r=0.277, P=0.0068), apo A-I (r=0.207, P=0.0435) and apo E (r=0.212, P=0.0388), and negatively with triglycerides (r=-0.228, P=0.0262). In the single nucleotide polymorphism of the SNP3 at the position -1131 nt of the apo A-V gene, apo A-V concentration was significantly higher in subjects with the T/T type than those with the C/C type (P<0.01), but not with the C/T type. In contrast, triglyceride concentration was significantly lower in subjects with the T/T type than in those with the C/C or C/T type (P<0.05). In addition, apo A-V concentration was much lower in non-insulin dependent diabetes mellitus than in healthy subjects (69.4 ± 44.3 ng/mL vs 179.2 ± 74.8 ng/mL, P<0.01). Our results show that plasma apo A-V concentration influences plasma triglyceride level, and that it is regulated by the polymorphism of the apo A-V gene. Plasma apo A-V concentrations are low in subjects with insulin resistance and diabetes mellitus.

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