Ultrasensitive and more specific enzyme immunoassay (immune complex transfer enzyme immunoassay) for p24 antigen of HIV-1 in serum using affinity-purified rabbit anti-p24 Fab′ and monoclonal mouse anti-p24 Fab′

✍ Scribed by Seiichi Hashida; Kazuya Hashinaka; Ichiro Nishikata; Atsushi Saito; Akihisa Takamizawa; Hideo Shinagawa; Eiji Ishikawa

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 517 KB
• Volume: 10
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/(sici)1098-2825(1996)10:5<302::aid-jcla11>3.0.co;2-0

✦ Synopsis

Previously, an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for p24 antigen of HIV-1 was developed. The immune complex comprising 2,4-dinitrophenyl-biotinyl-bovine serum albumin-rabbit anti-p24 Fab' conjugate, p24 antigen, and rabbit anti-p24 Fab'-p-Dgalactosidase conjugate was trapped onto polystyrene beads coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG, was eluted with &N-2,4-dinitrophenyI-~-lysine, and was transferred to polystyrene beads coated with streptavidin. p-D-Galactosidase activity bound to the streptavidin-coated polystyrene beads was assayed by fluorometry. This assay was highly sensitive. However, bound p-D-galactosidase activity had to be assayed for a long time (20 h), and the nonspecific signal was observed in 5% serum samples Ultrasensitive and More Specific Enzyme lmmunoassay (Immune Complex Transfer Enzyme lmmunoassay) for p24