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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

Overview
Journal J Clin Lab Anal
Publisher Wiley
Specialties Biomedical Engineering
Biotechnology
Pathology
Date 1998 Jul 22
PMID 9671175
Authors
S Ishikawa
S Hashida
K Hashinaka
A Adachi
S Oka
E Ishikawa
Affiliations
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Abstract

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).

References
1.
Ishikawa S, Hashida S, Hashinaka K, Hirota K, Saitoh A, Takamizawa A . Diagnosis of HIV-1 infection with whole saliva by detection of antibody IgG to HIV-1 with ultrasensitive enzyme immunoassay using recombinant reverse transcriptase as antigen. J Acquir Immune Defic Syndr Hum Retrovirol. 1995; 10(1):41-7. View
2.
Hashida S, Ishikawa S, Hashinaka K, Nishikata I, Oka S, Shimada K . Optimal conditions of immune complex transfer enzyme immunoassays for antibody IgGs to HIV-1 using recombinant p17, p24, and reverse transcriptase as antigens. J Clin Lab Anal. 1998; 12(2):98-107. PMC: 6807785. View
3.
Ishikawa S, Hashida S, Hashinaka K, Kojima M, Saito A, Takamizawa A . More sensitive immune complex transfer enzyme immunoassay for antibody IgG to p17 of HIV-1 with shorter incubation time for immunoreactions and larger volumes of serum samples. J Clin Lab Anal. 1997; 11(5):244-50. PMC: 6760719. DOI: 10.1002/(sici)1098-2825(1997)11:5<244::aid-jcla2>3.0.co;2-7. View
4.
Hashida S, Hashinaka K, Saitoh A, Takamizawa A, Shinagawa H, Oka S . Diagnosis of HIV-1 infection by detection of antibody IgG to HIV-1 in urine with ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens. J Clin Lab Anal. 1994; 8(4):237-46. DOI: 10.1002/jcla.1860080410. View
5.
Hashida S, Hashinaka K, Nishikata I, Oka S, Shimada K, Saitoh A . Measurement of human immunodeficiency virus type 1 p24 in serum by an ultrasensitive enzyme immunoassay, the two-site immune complex transfer enzyme immunoassay. J Clin Microbiol. 1995; 33(2):298-303. PMC: 227936. DOI: 10.1128/jcm.33.2.298-303.1995. View