Rapid formation of the immune complexes on solid phase in the immune complex transfer enzyme immunoassays for HIV-1 p24 antigen and antibody IgGs to HIV-1.
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'-beta-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 2,4-dinitrophenyl-HIV-1 antigen conjugates and subsequently with HIV-1 antigen-beta-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).
Department of Biochemistry, Miyazaki Medical College, Kiyotake, Japan.
S
S
Ishikawa
S
S
Hashida
K
K
Hashinaka
A
A
Adachi
S
S
Oka
E
E
Ishikawa
D000328:Adult; D000936:Antigen-Antibody Complex / Q000097:blood* / Q000276:immunology; D005260:Female; D015683:Gene Products, gag / Q000097:blood / Q000276:immunology; D015488:HIV Antigens / Q000097:blood* / Q000276:immunology; D016655:HIV Core Protein p24 / Q000097:blood* / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000097:blood / Q000276:immunology; D054303:HIV Reverse Transcriptase / Q000097:blood / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D011137:Polystyrenes; D012680:Sensitivity and Specificity; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus
D000936:Antigen-Antibody Complex; D015683:Gene Products, gag; D015488:HIV Antigens; D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D011137:Polystyrenes; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C054858:p17 protein, Human Immunodeficiency Virus Type 1; D054303:HIV Reverse Transcriptase