Enzyme immunoassay of the receptors for modified low density lipoprotein.
Mouse macrophages (line J 774) were incubated with monospecific goat anti-low density lipoprotein antibodies, which were conjugated to horseradish peroxidase (AB-HRP). Addition of low density lipoprotein (LDL) modified by treatment with malondialdehyde to cultures of these cells resulted in a dose-dependent increase in the amount of cell-associated enzyme activity. The concentration curve was hyperbolic with half-saturation of modified LDL at a concentration of about 3 micrograms/ml. This effect was completely blocked by polyinosinic acid and was not observed in experiments with human fibroblasts, which do not exhibit high affinity binding sites that recognize chemically modified LDL. Our data indicate that receptor-mediated endocytosis of AB-HRP in the presence of native or modified LDL may be used as very simple, efficient, and sensitive assay for investigation of the scavenger receptors for modified LDL.
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D008077:Lipoproteins, LDL / Q000276:immunology / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008315:Malondialdehyde; D051379:Mice; D011069:Poly I / Q000494:pharmacology; D011973:Receptors, LDL / Q000032:analysis*
D000906:Antibodies; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D011069:Poly I; D008315:Malondialdehyde; D006735:Horseradish Peroxidase