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Clinical Chemistry 46: 1638-1642, 2000;
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Right arrow Lipids, Lipoproteins, and Cardiovascular Risk Factors
(Clinical Chemistry. 2000;46:1638-1642.)
© 2000 American Association for Clinical Chemistry, Inc.

Articles

Determination of Apolipoprotein B-48 in Plasma by a Competitive ELISA

Anne-Marie Lorec1, Christine Juhel2, Yan Pafumi2, Henri Portugal1,3, Anne-Marie Pauli3, Denis Lairon2 and Catherine Defoort2,3,a

1 Laboratoire Central, Hôpital Sainte-Marguerite, 270 Bd Sainte-Marguerite BP 29, 13274 Marseille Cedex 09, France.

2 Unité INSERM 476, 18 Avenue Mozart, 13009 Marseille, France.

3 Laboratoire de Chimie Analytique, Faculté de Pharmacie, 27 Bd J Moulin, 13005 Marseille, France.
a Address correspondence to this author at: Unité INSERM 476, 18 Avenue Mozart, 13009 Marseille, France. Fax 33-491-75-15-62; e-mail defoort{at}marseille.inserm.fr

Background: Apolipoprotein B-48 (apoB-48) is produced by the small intestine, as part of chylomicrons, and appears to be a suitable marker for clinical studies of postprandial lipoproteins and related cardiovascular risk. Our aim was to develop, for routine analysis, an assay to quantify apoB-48 in plasma samples.

Methods: A microtiter plate was coated with a C-terminal apoB-48-specific heptapeptide. Plasma samples were incubated with appropriate detergent to allow competition between immobilized antigen and plasma apoB-48. Appropriate calibration curves were obtained in the ELISA, using calibrated lymph and chylomicrons.

Results: Treatment of plasma samples with the mild detergent Triton X-100 allowed an efficient competition between immobilized antigen and plasma apoB-48. No cross-reactivity was found with apoB-100, as checked by ELISA and Western blot analysis. Intra- and interassay CVs were 5.4% and 5.5%, respectively. In healthy subjects, apoB-48 concentrations markedly increased in the postprandial state, in parallel with triglycerides.

Conclusions: This new ELISA allows determination of the concentration of apoB-48 in normolipidemic plasma.




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