Apolipoprotein C species were separated in tubes, from human VLDL, by urea polyacrylamide gel electrophoresis. They were separately cut out and the gel slices containing them were used as antigens in rabbits. The resulting immunoglobulin G were tested for specificity. An apo C-III specific antibody was obtained. It was used to develop a competitive enzyme linked immunoassay (CELIA) for apo C-III content of acetone-delipi-dated human VLDL. Its effectiveness was compared with values of apo C content obtained from isoelectricfocusing (IEF) of the samples with content being estimated by densitometric measurements. A reasonable correlation (r = 0.67) of CELIA value for apo C-III with IEF-densitometric value was obtained. The assay showed no correlation with apo C-II values and thus may be useful for measuring the apo C-III isoforms. In addition the total apo C content and the distrbution of apo C-II, apo C-III0, apo C-III1, and apo C-III2 were measured by IEF densitmetry, for a small group of diabetics. These values were not different from those found for a group of non-diabetic controls. This method may be extended to allow measurement of apo E isoforms in the future.
|Date of Award||1985|