DETERMINATION OF PHENOBARBITAL BY THE MET HODS OF SOLID-PHASE ENZYME-IMMUNOASSAY - THE SELECTION OF AN OPTIMAL STRATEGY

Two variants (direct and indirect) of enzyme linked immunosorbent assa y (ELISA) of phenobarbital are compared. Both techniques were develope d on the basis of the same monoclonal antibodies, and horse radish per oxidase was used as the label in both cases. When microtitration plate s 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 a ntiphenobarbital antibodies, is preferable. In indirect ELISA, the sam ple volume was 5 mu l, the time of assay was 40 min, the variability c oefficient was <8%.