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Clinical Chemistry, Vol 33, 1163-1169, Copyright © 1987 by American Association for Clinical Chemistry
RH Neary and E Gowland
Free apolipoprotein A-1 (free A-1) is a low-molecular-mass complex of protein and lipid containing apolipoprotein A-1 (apo A-1). Using crossed immunoelectrophoresis, we separated free A-1 from the apo A-1 in high-density lipoproteins (HDL) and quantified free A-1 by comparison with a reference serum (containing 1.45 g of apo A-1 per liter) diluted in 9 mol/L urea solution. This latter treatment yields apo A-1 containing protein-lipid complexes of the same size and electrophoretic mobility as free A-1. Within-day precision (CV), determined by replicate analysis of two samples with mean free A-1 concentrations of 48 and 138 mg/L, was 9.1 and 7.2%, respectively. We also showed that the concentration of free A-1 is not stable in serum or plasma either at 4 degrees C or when frozen. The mean concentration of free A-1 in 28 fasted, healthy subjects was 75.3 (SD 13.6) mg/L. The postprandial increase was not statistically significant. The percentage of total apo A-1 in the free form in serum ranged from 3.5% to 8.1%, less than the 10% to 30% reported by others who used radial immunodiffusion to measure free A-1. Because radial immunodiffusion does not separate free A-1 from HDL, we believe that that technique overestimates free A-1. We also used crossed immunoelectrophoresis to measure free A-1 in 76 hyperlipidemic patients. Those with Fredrickson types III and V had significantly increased concentrations of free A-1 (P less than .0001). Correlations between free A-1 and cholesterol and triglycerides in serum were significant (P less than .005 and P less than .001, respectively). Possible roles for free A-1 in lipid metabolism are discussed.
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