Immunoturbidimetry of serum apolipoproteins A-I and B on the Cobas Bio centrifugal analyzer: method validation and reference values

Abstract

We describe an immunoturbidimetric assay for estimating the concentration of apolipoproteins (apo) A-I and B in serum with a Cobas Bio centrifugal analyzer. Several analytical variables were studied: pH, temperature, wavelength, reaction time, sample pretreatment, antisera dilution, concentration of NaCl, phosphate buffer, and PEG 6000; a good optical response was obtained with small volumes of sample and antisera. Imprecision was low, with intrarun coefficient of variance (CV) from 1.1 to 2.8% and interrun CV from 1.8 to 3.2%. We demonstrated a good correlation between our method and radial immunodiffusion for apo A-I (r = 0.975) and apo B (r = 0.980). Reference values were calculated from a sample of 494 healthy subjects, and separate intervals for the ratio of apo A-I to apo B in gender and age subgroups were established.