Immunochemical Determination of Apolipoprotein A-I and B in Hyperlipidemic Subjects

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Serum concentration of apolipoprotein A-I (apo A-I) and B (apo B) was determined by rocket immunoelectrophoresis. The monospecific antiserum for the assay was prepared in the rabbits by immunizing with isolated and purified apo A-I or LDL. The supporting medium for the electrophoresis was prepared with 1% agarose-gel which contained 1.5% anti-apo A-I or 2% anti-apo B. Serum samples were carbamylated before electrophoresis. A normal serum usually required 1: 12 dilution for apo A-I and 1: 6 dilution for apo B. The electrophoresis was carried out at constant voltage of 2V/cm for 16 hours at 15°C. The buffer was 0.02M veronal at pH 8.6. The immunoprecipitates were stained with amido black 10B and measured the hights. The apoprotein concentrations were expressed as arbitrary units/100ml where the serum used for references assumed to contain 100 units/100ml of each apoprotein.<BR>Apo A-I and apo B levels in 9 young healthy males revealed 102 ± 18 units/100ml and 99 ± 29 units/100ml, respectively. In 11 young females, apo A-I and apo B were 111 ± 7 and 102 ± 21 units/100ml, respectively (TABLE I). Increased levels of apo B was observed in hyperlipidemic subjects. In 7 patients with type II hyperlipidemia mean apo B lavel was 203± 26 units/100ml and in 16 type IV patients the level was 208 ± 16 units/100ml. These values were significantly higher than 18 controls with apo B concentration of 124 ± 24 units/100ml. Apo A-I in hyperlipidemics showed slight decrease in type II patients but no significant changes in type IV cases (TABLE II).<BR>Serum concentration of apo B significantly correlated with the cholesterol levels. On the other hand, apo A-I levels correlated with cholesterol concentration in normolipidemic subjects but the correlation was inversed in hyperlipidemics (TABLE III). These observation suggested that in hyperlipidemics the serum apo A-I was relatively deficient for their cholesterol content.
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Immunochemical Determination of Apolipoprotein A-I and B in Hyperlipidemic Subjects

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