Rapid formation of the immune complexes on solid phase in the immune complex transfer enzyme immunoassay for HIV‐1 p24 antigen and antibody IgGs to HIV‐1

摘要

In order to perform the immune complex transfer enzyme immunoassays for HIV‐1 p24 antigen and antibody IgGs to HIV‐1 p17, reverse transcriptase and gp41 antigens as rapidly as possible, methods for rapid formation of the immune complexes on solid phase are described. HIV‐1 p24 antigen was reacted with monoclonal anti‐p24 Fab′‐β‐ ‐galactosidase conjugate at a high concentration and subsequently with polystyrene beads coated successively with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG and 2,4‐dinitrophenyl‐biotinyl bovine serum albumin‐affinity‐purified anti‐p24 Fab′ conjugate. Antibody IgGs to HIV‐1 were reacted with polystyrene beads coated successively with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG and 2,4‐dinitrophenyl‐HIV‐1 antigen conjugates and subsequently with HIV‐1 antigen‐β‐D‐galactosidase conjugates. The periods of time used for the formation of the immune complexes comprising the three components on the polystyrene beads (15–30 min) were much shorter than those used in the previous immune complex transfer enzyme immunoassays (90–300 min), and the sensitivities of the present and previous immune complex transfer enzyme immunoassays were similar. The detection limit of the HIV‐1 p24 antigen by the present and previous methods were similar (3 to 10 zmol/assay). J. Clin. Lab. Anal. 12:227–237, 1998. © 1998 Wiley‐Liss, Inc.