Time-resolved fluorometric assay for measuring cell binding and association of native and oxidized low-density lipoproteins to macrophages

Europium-labeled native and oxidized low-density lipoprotein (LDL) were used to measure their binding and cell association to mouse peritoneal macrophages, to suspended human monocyte cell line THP-1 cells, and to differentiated THP-1 macrophages. Cell binding and association were concentration dependent and saturable and showed the characteristics of Ligand-receptor interaction The validity of this assay was also supported by comparison with the method using (125)iodine-labeled LDL. This nonradioactive assay proved to be specific, sensitive and simple and avoided any potential lipid peroxidation of LDL brought about by labeling lipoproteins with the widely used radioactive iodine. The latter fact is very important in studying lipoprotein-receptor interactions. (C) 1999 Academic Press.