Background: Apolipoprotein B-48 (apoB-48) is produced by the small intestin
e, as part of chylomicrons, and appears to be a suitable marker far clinica
l studies of postprandial lipoproteins and related cardiovascular risk. Our
aim was to develop, for routine analysis,an assay to quantify apoB-48 in p
lasma samples.
Methods: A microtiter plate was coated with a C-terminal apoB-48-specific h
eptapeptide. Plasma samples were incubated with appropriate detergent to al
low 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 a
llowed an efficient competition between immobilized antigen and plasma apoB
-48. No cross-reactivity was found with apoB-100, as checked by ELISA and W
estern blot analysis. Intra- and interassay CVs were 5.4% and 5.5%, respect
ively. In healthy subjects, apoB-48 concentrations markedly increased in th
e postprandial state, in parallel with triglycerides.
Conclusions: This new ELISA allows determination of the concentration of ap
oB-48 in normolipidemic plasma. (C) 2000 American Association for Clinical
Chemistry.