A sandwich enzyme-linked immunosorbent assay for human serum paraoxonase concentration

Abstract

Serum paraoxonase (PON) is associated with plasma high density lipoproteins, and prevents the oxidative modification of low density lipoproteins. We have developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA), using two monoclonal antibodies against PON, to measure serum PON concentration. The concentration of PON in healthy Japanese subjects was 59.3 ± 1.3 μg/mL (mean ± SEM; n = 87). Serum PON concentrations in relation to the PON 192 genetic polymorphism were: 69.5 ± 2.9 μg/mL in the QQ genotype; 63.0 ± 1.9 μg/mL in the QR genotype; and 52.8 ± 1.7 μg/mL in the RR genotype. Concentrations were significantly lower in the RR than in the QQ genotype (P < 0.01). Serum paraoxonase specific activity was higher in RR than in QQ subjects (18.6 ± 0.40 vs. 2.56 ± 0.05 nmol/min/μg, P < 0.01), but arylesterase specific activity was unrelated to genotype. PON concentration was positively associated (P < 0.001) with both serum arylesterase activity and, after adjusting for the effect of the position 192 polymorphism, with serum paraoxonase activity. Subjects with angiographically verified coronary heart disease had significantly lower PON concentrations than the healthy controls (52.0 ± 2.3 μg/mL; n = 35, P < 0.01). This association was independent of the position 192 genotype. Our new ELISA should be of value for epidemiologic and clinical studies of serum PON concentration. —Kujiraoka, T., T. Oka, M. Ishihara, T. Egashira, T. Fujioka, E. Saito, S. Saito, N. E. Miller, and H. Hattori. A sandwich enzyme-linked immunosorbent assay for human serum paraoxonase concentration. J. Lipid Res. 2000. 41: 1358–1363.

Keywords

• low density lipoproteins
• high density lipoproteins
• phospholipids
• coronary heart disease