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Clinical Chemistry 48: 884-890, 2002;
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Right arrow Lipids, Lipoproteins, and Cardiovascular Risk Factors
(Clinical Chemistry. 2002;48:884-890.)
© 2002 American Association for Clinical Chemistry, Inc.

Quantitative Measurement of Lipoprotein Particles Containing Both Apolipoprotein AIV and Apolipoprotein B in Human Plasma by a Noncompetitive ELISA

Fanny Ferrer1,2, Edith Bigot-Corbel2, Patrick N'Guyen3, Michel Krempf1 and Jean-Marie Bard1,2a

1 INSERM U539, Centre de Recherche en Nutrition Humaine, CHU Hôtel Dieu, 44035 Nantes, France.

2 Laboratoire de Biochimie, UFR de Pharmacie, 44035 Nantes, France.

3 Ecole Nationale Vétérinaire, 44300 Nantes, France.

aAddress correspondence to this author at: Laboratoire de Biochimie Fondamentale et Appliquée, UFR de Pharmacie, 1 rue Gaston Veil, 44093 Nantes Cédex 1, France. E-mail Jean-Marie.Bard{at}sante.univ-nantes.fr.

Background: A reliable method for plasma would be useful to investigate the role of apolipoprotein (apo) AIV when associated with apo B-containing or triglyceride-rich lipoproteins.

Method: We used a sandwich ELISA to quantify lipoprotein B:AIV particles (Lp B:AIVf; lipoproteins containing at least apo B and apo AIV) in plasma. The method used microtiter plates coated with purified anti-apo B immunoglobulins that selectively retained apo B-containing particles. Lipoproteins containing both apo B and apo AIV were distinguished from those containing only apo B by use of a peroxidase-labeled anti-apo AIV antibody. These subspecies were revealed by ABTS® reagent and further quantified by spectrophotometry. Results were expressed in mg/L apo AIV associated with apo B. This method was applied to samples with different cholesterol and triglyceride concentrations.

Results: The developed sandwich ELISA method identified and quantified Lp B:AIVf in plasma samples. Within- and between-run CVs were ~10%, and analytical recoveries were 95–107%. Results were not significantly influenced by addition of triglycerides or by storage at -20 °C (up to 9 months). Under these conditions, plasma Lp B:AIVf concentrations were statistically higher in hypercholesterolemic and mixed hyperlipidemic individuals (53 ± 13 mg/L; P <0.001 and 70 ± 18 mg/L; P <0.001, respectively) than in normolipidemic individuals (43 ± 12 mg/L). Lp B:AIVf concentration appeared to be well correlated with total cholesterol, triglycerides, LDL-cholesterol, and apo B. These results were in contrast to total apo AIV, which was not different between dyslipidemic and normolipidemic individuals.

Conclusions: The developed ELISA method for Lp B:AIVf in plasma combines specificity, reliability, and speed. The increase in Lp B:AIVf concentrations in various dyslipidemic states, together with a lack of change in total apo AIV concentrations, suggests a redistribution of apo AIV toward apo B-containing lipoproteins when these lipoproteins accumulate.






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Copyright © 2002 by the American Association for Clinical Chemistry.