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Clinical Chemistry 34: 739-743, 1988;
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Clinical Chemistry, Vol 34, 739-743, Copyright © 1988 by American Association for Clinical Chemistry

Quantification of human apolipoprotein A-IV by "sandwich"-type enzyme- linked immunosorbent assay

M Rosseneu, G Michiels, W De Keersgieter, J Bury, JP De Slypere, H Dieplinger and G Utermann
Department of Clinical Chemistry, A.Z. St-Jan, Ruddershove, Brugge, Belgium.

A specific and sensitive "sandwich"-type enzyme-linked immunosorbent assay (ELISA) has been developed for quantifying human apo A-IV. Using apo A-IV immunosorbent columns, we isolated monospecific anti-apo A-IV antibodies for coating the ELISA plates and for preparing peroxidase- antibody conjugate. The assay can detect as little as 0.20 ng of apo A- IV, with mean intra- and interassay CVs of 3.6% and 8.2%, respectively. The apoA-IV concentrations in normolipemic and hyperlipemic plasma were unaffected by either delipidation or treatment with detergents or urea. To validate the ELISA assay we compared it with an immunoelectrophoretic technique. ApoA-IV concentrations in plasma from normo- and dyslipemic subjects compared well by the two assays (r = 0.89). The mean apo A-IV concentration, measured by ELISA in plasma from 50 normolipemic subjects, was 143 (SD 52) mg/L; values for dyslipemic subjects were not significantly different. We also used this new assay to monitor apo A-IV profiles of normolipemic and hypertriglyceridemic plasma after chromatographic fractionation.


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Copyright © 1988 by the American Association for Clinical Chemistry.