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

✍ Scribed by S.N. Preobrazhensky; V.O. Ivanov; I.V. Fuki; V.P. Tsibulsky; A.M. Kameneva; V.S. Repin; G.A. Ermolin

• Publisher: Elsevier Science
• Year: 1985
• Tongue: English
• Weight: 495 KB
• Volume: 149
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(85)90504-4

✦ Synopsis

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 lo-20 times more effectively than reductively methylated LDL or native LDL in the presence of heparin. Receptor-specific K,,, and maximum binding capacity calculated from these data were 5.4 pg/ ml and 177.5 ng/mg of cell protein, respectively. Receptor-specific K,,, and maximum binding capacity for surface '*'I-LDL binding measured in the same experiment were 1 I.0 &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 hpoproteins. 0 1985 Academic press, Inc.