[Determination of phenobarbital by solid-phase immunoenzyme analysis. Choice of optimal strategy]

Two variants (direct and indirect) of enzyme linked immunosorbent assay (ELISA) of phenobarbital are compared. Both techniques were developed on the basis of the same monoclonal antibodies, and horse radish peroxidase was used as the label in both cases. When microtitration plates are used as the solid phase, indirect ELISA, in which phenobarbital of the sample competes with phenobarbital sorbed on plates in the form of a conjugate with protein for the binding with peroxidase-labeled antiphenobarbital antibodies, is preferable. In indirect ELISA, the sample volume was 5 microliters, the time of assay was 40 min, the variability coefficient was < 8%.