ENHANCEMENT OF ELISA SENSITIVITY FOR THE DETECTION OF RABBIT ANTI-HIVANTIBODIES USING APAAP COMPLEX

The use of an enzyme-antienzyme method in ELISA is shown to increase s ensitivity. This modification of ELISA using APAAP complex was employe d for the detection of anti-HIV antibodies. The APAAP complex (immune complex of seal alkaline phosphatase with mouse monoclonal antibody ag ainst this enzyme) produced by interaction of crude alkaline phosphata se with culture medium containing monoclonal antibody to the phosphata se was used in the analysis without further purification and had a low er background than the conjugate of protein A and phosphatase bound to each other by glutaraldehyde. Optimal concentrations of protein A and rabbit anti-mouse IgG antiserum and composition and dilution of the A PAAP complex that provide a maximum sensitivity of the method were det ermined. The use of the enzyme-antienzyme method as compared with trad itional ELISA methods enabled us to increase the sensitivity of the an alysis for the detection of rabbit anti-HIV antibodies 4- to 16-fold.