Characterization and quantitation of the apoproteins of high-density lipoprotein by capillary electrophoresis

A method has been developed using capillary electrophoresis (CE) to quantitate plasma levels of apoprotein A-I (apoA-I) and apoprotein A-II (apoA-II) in high-density lipoprotein (HDL) samples. ApoA-I and apoA-II are resolved by CE in delipidated and non-delipidated HDL samples. Concentrations of apoA-I and apoA-II were calculated from their peak areas in the electropherogram. Results of the analysis of Sigma plasma standards (Controls 1 and 2) using CE are in good agreement with values obtained by Sigma using immunoturbidimetric assay. CE and reverse-phase high-performance liquid chromatography (RP-HPLC) were found to be complementary in the study of apoA-I and apoA-II. RP-HPLC resolves the isoforms of purified apoA-I and apoA-II, but it cannot resolve mixtures of them because the retention times of the isoforms overlap. CE separates apoA-I from apoA-II, but it does not resolve the isoforms. Matrix-assisted laser desorption/ ionization mass spectrometry was used to identify the isoforms of apoA-I and apoA-II by their molecular weight (M(r)) in fractions collected from RP-HPLC.