Abstract
The use of an enzyme-antienzyme method in ELISA is shown to increase sensitivity. This modification of ELISA using APAAP complex was employed for the detection of anti-HIV antibodies. The APAAP complex (immune complex of seal alkaline phosphatase with mouse monoclonal antibody against this enzyme) produced by interaction of crude alkaline phosphatase with culture medium containing monoclonal antibody to the phosphatase was used in the analysis without further purification and had a lower 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 APAAP complex that provide a maximum sensitivity of the method were determined. The use of the enzyme-antienzyme method as compared with traditional ELISA methods enabled us to increase the sensitivity of the analysis for the detection of rabbit anti-HIV antibodies 4- to 16-fold.
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