Loss of A-apoprotein immunoreactivity during high-density lipoprotein separation

By use of an electroimmunoassay, concentrations of A-apoproteins were estimated in serum and in corresponding apoprotein fractions isolated by ultracentrifugation. These values were compared with high-density lipoprotein concentrations determined by analytical ultracentrifugation. Concentrations of A-apoproteins estimated in serum were considerably higher than in isolated high-density lipoprotein fractions. These discrepancies could not be accounted for entirely by material losses into other fractions during ultracentrifugal fractionation. No comparable differences in apoprotein-B concentrations were observed during the ultracentrifugal separation of low-density lipoprotein. Concentrations of A-apoproteins estimated in the residual serum after precipitation of low-density lipoproteins by heparin and manganous ions were also lower than in the corresponding whole sera. The discrepancies persisted after treatment of serum and isolated fractions with tetramethylurea, urea (9 mol/l), and by heating at 52 degrees C for 3 hours. It is considered that separation by ultracentrifugation induces subtle alterations in the surface structure of the lipoprotein species which give rise to changes in immunoreactivity.