Postprandial determination of Apo B-48 levels in whole plasma of healthy young individuals by a double-sandwich ELISA*

Autores: Mantilla Mora Gerardo, Sierra Ariza Iván Darío, Mendivil Cárdenas Carlos Olimpo, Pérez Sánchez Clara Eugenia

Resumen

High postprandial concentrations of chylomicrons and its remnants are correlated with an atherosclerosis progression. Apolipoprotein B-48 is an essential component of these lipoproteins and appears to be a suitable marker for clinical studies of postprandial lipid metabolism and its relationship to cardiovascular risk. A double-sandwich ELISA was developed for routine Apo B-48 quantification in untreated human plasma. The postprandial behavior of Apo B-48 in healthy young individuals was described employing the method, and correlations between plasma Apo B-48 and lipid and clinical parameters. A polyclonal antibody directed against the carboxy-terminal extreme of Apo B-48 was developed, and appropriate calibrators were prepared using an affinity column to which the developed antibody was attached. A mouse monoclonal antibody able to recognize both Apo B-48 and Apo B-100 was used as second antibody, and an enzyme coupled anti-mouse IgG was used as third antibody. The plasma Apo B-48 levels were determined in 42 healthy young individuals of both sexes in the fasting state and hourly after the consumption of a breakfast with 40 g fat. The technique showed an intra-assay variation of 3,0-3,8%. Plasma Apo B-48 fluctuated between 0,5 and 0,8 mg/L in the fasting state, with an hourly increase to reach a maximum between 4,6 and 8,4 mg/L at the third postprandial hour. The postprandial Apo B-48 area under curve and third hour Apo B-48 showed a strong correlation with body mass index (r=0,58 and 0,8 respectively). This paper presents a novel assay that makes Apo B-48 quantification easier and faster with adequate precision and without requirements for sample processing. Plasma Apo B-48 in healthy young individuals showed postprandial kinetics characterized by low fasting concentrations that increase to a peak about 3 hours after a meal and return to low values after 6 hours.

Palabras clave: Apo B-48 postprandial lipemia cardiovascular risk immunoassay chylomicrons lipoproteins.

2003-12-23   |   1,568 visitas   |   Evalua este artículo 0 valoraciones

Vol. 6 Núm.18. Diciembre 2003 Pags. 130-136 MedUNAB 2003; 6(18)