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

✍ Scribed by Setsuko Ishikawa; Seiichi Hashida; Kazuya Hashinaka; Akio Adachi; Shinichi Oka; Eiji Ishikawa

• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 89 KB
• Volume: 12
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/(sici)1098-2825(1998)12:4<227::aid-jcla7>3.0.co;2-4

✦ Synopsis

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-D-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 2,4dinitrophenyl-HIV-1 antigen conjugates and subsequently with HIV-1 antigen-β-Dgalactosidase 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).