Iy. Sakharov et al., ENHANCEMENT OF ELISA SENSITIVITY FOR THE DETECTION OF RABBIT ANTI-HIVANTIBODIES USING APAAP COMPLEX, Analytica chimica acta, 319(1-2), 1996, pp. 265-269
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.