Clinical Chemistry
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Clinical Chemistry 27: 249-252, 1981;
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Clinical Chemistry, Vol 27, 249-252, Copyright © 1981 by American Association for Clinical Chemistry

Double-antibody fluorescence immunoassay of tobramycin

HT Karnes, JC Gudat, CM O'Donnell and JD Winefordner

This heterogeneous assay for tobramycin involves fluorescein-labeled tobramycin, which competes with native unlabeled tobramycin for anti- tobramycin binding sites. Bound and free labeled antigen are separated by precipitation with a second antibody. Fluorescence intensity of the resuspended precipitate is inversely proportional to native tobramycin concentration. Background interference was consistently about 10% of the total fluorescence precipitated. Assay sensitivity was sufficient to detect nanogram quantities of tobramycin per assay tube. Correlation coefficients (r) were 0.96 and 0.98 for comparisons of this assay with a microbiological assay and a radioimmunoassay, respectively. Mean analytical recovery was 101% and the CV was less than 10% throughout the therapeutic range.




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Copyright © 1981 by the American Association for Clinical Chemistry.