The clinical relevance of apoE concentration in lipoprotein fractions should be evaluated. We investigated the impact of the common apolipoprotein (apo) E polymorphism in conjunction with very low density lipoprotein (VLDL) apoE concentration on the receptor binding properties of VLDL preparations from 17 normolipidemic subjects of the HepG2 cell surface receptors. All six apoE genotypes were studied. When apoE genotype alone was considered, two subgroups could be distinguished: VLDL without apoE isoform E2 (VLDL-3/3, VLDL-3/4, and VLDL-4/4) showed significantly higher affinity than VLDL with apoE2 (VLDL-4/2, VLDL-3/2, and VLDL-2/2). Once we adjusted for VLDL apoE content, we observed that VLDL affinity to HepG2 cell surface receptors decreased, according to apoE genotype, in the following order: VLDL-4/4 (100%) > VLDL-3/4 (93%) > VLDL-3/3 (82%) > VLDL-4/2 (53%) > VLDL-3/2 (36%) > VLDL-2/2 (30%). Moreover, we found that VLDL apoE concentration could modify isoform-specific binding. An analysis in 47 subjects showed that the concentration of total VLDL protein and the VLDL apoE concentration varied considerably. The variation of VLDL apoE was independent of apoE genotype and corresponding serum apoE levels. We conclude that, in addition to apoE genotype, apoE content of VLDL is an important determinant of the receptor binding properties of VLDL.