Clinical Chemistry
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Clinical Chemistry 30: 1174-1178, 1984;
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Clinical Chemistry, Vol 30, 1174-1178, Copyright © 1984 by American Association for Clinical Chemistry

Ligand displacement immunoassay: a novel enzyme immunoassay demonstrated for measuring theophylline in serum

JA Hinds, CF Pincombe, RK Kanowski, SA Day, JC Sanderson and P Duffy

We describe a ligand displacement immunoassay for measurement of theophylline in serum or plasma and show it to be clinically useful. A conjugate of theophylline-specific antiserum covalently bound to micro- crystalline cellulose and a beta-galactosidase-labeled theophylline derivative is prepared as a lyophilized reagent. Sample is added to the conjugate and, after 5-min incubation at room temperature, the bound and displaced enzyme-ligand are separated by brief centrifugation. The enzyme activity of the displaced enzyme-ligand is measured on a spectrophotometer or centrifugal analyzer. Linear calibration curves are obtained with appropriate sample dilution, allowing generation of the standard curve with a saline zero reference and a single calibrator. Performance of the assay compares favorably with that of other theophylline assays; comparison studies with use of 145 clinical specimens gave a coefficient of correlation with EMIT of 0.97.




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Copyright © 1984 by the American Association for Clinical Chemistry.