The design of immunogens for raising antisera containing antibodies specifically directed against paracetamol is discussed. A derivative of the drug with minimal structural alteration was synthesised, coupled to the carrier protein key-hole limpet haemocyanin, and used to immunise sheep. The evaluation of the resulting antisera is described. Preparation of a fluorescent label with reduced bridge-binding then allowed the development of a polarisation fluoroimmunoassay for paracetamol in buffer, suitable for conversion to a direct serum assay.
SUMMARY.A specific polarisation fluoroimmunoassay for the measurement of paracetamollevels in serum has been developed for use in emergency toxicological screening. It is based on the use of a fluorescein-labelled analogue and a sheep antiserum and exploits the rapid dissociation kinetics of the hapten-antibody complex to enable the label and antiserum to be combined as a single reagent. Paracetamollevels are determined by adding 5 I1L of serum to 1·5 mL of the single reagent, incubation at ambient temperature for a few minutes and measurement of fluorescence polarisation. In addition to its speed and simplicity, the assay is both accurate and precise and the results obtained correlate closely with those from the commonly used chemical and enzymatic techniques.
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