Enzyme-linked immunosorbent assay to measure apolipoproteins Al and B secreted by a human hepatic carcinoma cell line (Hep G2)

✍ Scribed by Susan E. Hahn; Joel G. Parkes; David M. Goldberg

• Book ID: 102877670
• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 694 KB
• Volume: 6
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860060404

✦ Synopsis

We describe an enzyme-linked immunosorbent assay (ELISA) to measure apolipoproteins Al and B secreted by Hep G2 cells and in cell homogenates. These assays utilize comercially available polyclonal antibodies, affinity-purified to improve their specificity, thereby achieving a dramatic increase in the sensitivity of the assay. These affinity-purified antibodies were also more sensitive than a series of monoclonal antibodies tested. We achieved a sensitivity of 0.4 ng in the apo Al assay, and a sensitivity of 5 ng in the apo B assay. By these methods, we measured secretion rates by Hep G2 cells of 358 k 41 ngimg cell protein/hr for apo B and 137 * 8 n g h g cell proteinlhr for apo Al. These assays also allowed the measurement of intracellular apolipoproteins and thus can be used to facilitate investigations of human lipoprotein metabolism in cell culture systems.