Enzyme-linked immunoreceptor assay of low-density-lipoprotein receptors

A new modification of enzyme immunoassay-enzyme-linked-immunoreceptor assay (ELIRA) was used to study low-density lipoprotein (LDL) binding by cultured human skin fibroblasts. Time-course and concentration curves obtained by this method were typical of LDL binding by cultured fibroblasts. According to ELIRA, fibroblasts bound native LDL 10-20 times more effectively than reductively methylated LDL or native LDL in the presence of heparin. Receptor-specific Km and maximum binding capacity calculated from these data were 5.4 micrograms/ml and 177.5 ng/mg of cell protein, respectively. Receptor-specific Km and maximum binding capacity for surface 125I-LDL binding measured in the same experiment were 11.0 micrograms/ml and 138.0 ng/mg of cell protein. Incubation of cells with isolated LDL or with unfractionated serum containing the same amount of apoB yielded similar concentration curves for lipoprotein binding. These data indicate that the ELIRA can be used for investigation of receptor-mediated lipoprotein binding without purification of lipoproteins.