TY - JOUR
T1 - Analysis of the HIV-1 gp41 specific immune response using a multiplexed antibody detection assay
AU - Opalka, David
AU - Pessi, Antonello
AU - Bianchi, Elisabetta
AU - Ciliberto, Gennaro
AU - Schleif, William
AU - McElhaugh, Michael
AU - Danzeisen, Renee
AU - Geleziunas, Romas
AU - Miller, Michael
AU - Eckert, Debra M.
AU - Bramhill, David
AU - Joyce, Joseph
AU - Cook, James
AU - Magilton, William
AU - Shiver, John
AU - Emini, Emilio
AU - Esser, Mark T.
PY - 2004/4
Y1 - 2004/4
N2 - A fluorescence-based, multiplexed, antibody-binding and mapping assay was developed to characterize antibody responses in HIV-1-infected individuals to the ectodomain of the HIV-1 gp41 envelope glycoprotein. The antigen panel included intact recombinant gp41, the fusion peptide region, the polar region, the N-heptad region, the C-heptad region as well as overlapping epitopes in the 2F5 and 4E10 monoclonal antibody-binding regions. The panel included both native and constrained peptides specifically designed to mimic putative gp41 prefusion and fusion intermediates. The results of these analyses revealed a broad pattern of immune responses against the test antigens, suggesting that none of these gp41 regions are immunologically silent. The HIV-1-positive sera were also evaluated using infectivity inhibition assays. No correlation was evident between the breadth or magnitude of specific anti-gp41 reactivities and virus neutralization potency. These evaluations demonstrated the substantial potential of the multiplexed antibody binding and mapping assay for rapid and sensitive analysis of complex antibody responses.
AB - A fluorescence-based, multiplexed, antibody-binding and mapping assay was developed to characterize antibody responses in HIV-1-infected individuals to the ectodomain of the HIV-1 gp41 envelope glycoprotein. The antigen panel included intact recombinant gp41, the fusion peptide region, the polar region, the N-heptad region, the C-heptad region as well as overlapping epitopes in the 2F5 and 4E10 monoclonal antibody-binding regions. The panel included both native and constrained peptides specifically designed to mimic putative gp41 prefusion and fusion intermediates. The results of these analyses revealed a broad pattern of immune responses against the test antigens, suggesting that none of these gp41 regions are immunologically silent. The HIV-1-positive sera were also evaluated using infectivity inhibition assays. No correlation was evident between the breadth or magnitude of specific anti-gp41 reactivities and virus neutralization potency. These evaluations demonstrated the substantial potential of the multiplexed antibody binding and mapping assay for rapid and sensitive analysis of complex antibody responses.
KW - Epitope-mapping
KW - Human immunodeficiency virus
KW - Luminex
KW - Multiplexed
KW - Neutralizing antibody
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U2 - 10.1016/j.jim.2004.01.016
DO - 10.1016/j.jim.2004.01.016
M3 - Article
C2 - 15099755
AN - SCOPUS:11144358557
VL - 287
SP - 49
EP - 65
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -