Quantification of human serum paraoxonase by enzyme-linked immunoassay: population differences in protein concentrations

• Published in: Biochemical journal Vol. 304 ( Pt 2); no. 2; pp. 549 - 554
• Main Authors: Blatter Garin, M C, Abbott, C, Messmer, S, Mackness, M, Durrington, P, Pometta, D, James, R W
• Format: Journal Article
• Language: English
• Published: England 01.12.1994
• Subjects: Animals; Antibodies, Monoclonal; Apolipoprotein A-I - metabolism; Aryldialkylphosphatase; Edetic Acid - pharmacology; Egtazic Acid - pharmacology; Electrophoresis, Gel, Two-Dimensional; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - statistics & numerical data; Esterases - blood; Female; Humans; Immunoblotting; Lipids - blood; Lipoproteins, HDL - blood; Male; Microchemistry; Rats; Reference Values
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/bj3040549

Paraoxonase is a serum protein bound to high-density lipoproteins (HDLs). The physiological function of the enzyme is unknown, but a role in lipid metabolism has been postulated. To date, studies of the protein have had to rely on measurements of enzyme activity with various substrates. We have developed a highly specific, competitive e.l.i.s.a. using a previously characterized monoclonal antibody. The assay can detect 20 ng of paraoxonase with a working range of 75-600 ng. Intra- and interassay coefficients of variation were 6.5 and 7.9% respectively. Serum concentrations of paraoxonase in healthy subjects from Geneva and Manchester ranged from 25 to 118 micrograms/ml. There were significant differences in mean concentrations between the two groups (Geneva, 79.3 +/- 18.7 micrograms/ml; Manchester, 59.9 +/- 24.1 micrograms/ml: P < 0.001), differences also apparent when subjects were compared according to paraoxonase phenotype. These appeared to be largely a consequence of differences in apolipoprotein A-I concentrations between the two populations, suggesting that HDL particle number may be important in determining serum levels of paraoxonase. Paraoxonase specific activities were also significantly different between the two groups of subjects (Geneva, 2.08 +/- 0.96 units/mg; Manchester, 3.08 +/- 1.73 units/mg: P < 0.001), which may reflect differences in HDL particle composition. The e.l.i.s.a. should furnish the necessary complement to studies of paraoxonase enzymic activity and has already provided evidence for differences with respect to serum levels of the protein both between populations and between phenotypes within populations.