Biochemistry 3386B Lecture Notes - Therapeutic Drug Monitoring, Immunoassay, Die Shrink
27 Apr 2013
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Automated assays (done in the core labs: homogeneous competitive binding immunoassays. The most common automated assays for therapeutic drug monitoring and toxicology. A labeled drug corresponding to the one in the patient sample is added. Label: fluorescent compound (fpia; fluorescence polarization immunoassay) Best method: particle (petia; particle enhanced turbimetric inhibition immunoassay) Antibody bound drug-linked particles form insoluble aggregates. Tag= spherical latex particle (40nm dia- turbidity density: enzyme (emit; enzyme mediated immunoassay technique) Any drug-linked enzyme which becomes bound to the antibody is inactivated. Residual amount (unbound) enzyme activity is proportional to the concentration of drug in the patient sample (observe nad + nadh conversion at 340nm) because free has activity and bound does not. The unlabeled drug in the sample and the labeled drug are added to the sample and compete for the binding site on the antibody (binding protein) Note: antibody is against that drug (monoclonal) and is limited to the amount needed to bind the drug.
