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Immunoassay of drugs and toxins[DOSAGE DES MEDICAMENTS ET DES TOXIQUES PAR IMMUNO-ANALYSE]
The simultaneous presence of drug and its metabolites in the same biological sample presents the problem of specificity of the drug immunoassay. The knowledge of the drug pharmacology is an important factor for determining the site of attachment of the drug to the carrier protein. In the function of this selected site of attachment and the selection of appropriate monoclonal or polyclonal antibodies, the drug antibody specificity can be adapted to drug monitoring or pharmacokinetic or toxicological studies. The physical separation by solvants or by chromatographical procedures and the definition of appropriate incubation conditions for the drug immunoassay can explain the lack of specificity of drug antibodies.
Cross-reactivity in immunoanalysis. The lack of specificity is a major limitation in immunoanalysis. Cross-reactivity dependson the type of antibodies used (monoclonal, polyclonal), assay configuration, and relative concentrations of cross-reactive compounds. Cross-reactivity may be evaluated by different methods, which are presented and discussed.
Different mecanisms can be involved in analytical drug interference in immunoassays : among them, specificity problems are the most frequently encountered. Antibody specificity can be reported in two manners, each of them presenting its own interest. Heparin therapy interference on free T4 assay represents a particular case, discussed separately.
Thevetin B, a cardiac glycoside of Thevetia neriifolia Juss. seeds, was determined in serum by fluorescence polarization immunoassay. Anti-digitoxin antibody was used, thevetin B genin being structurally identical to digitoxigenin. Cross-reactivity of 94% was found by this method, for concentrations from 5 to 80 ng ml-1.
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