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J Immunol Methods. 2013 Nov 29;397(1-2):47-54. doi: 10.1016/j.jim.2013.09.003. Epub 2013 Sep 13.

Isolation of HIV-1-reactive antibodies using cell surface-expressed gp160Δc(BaL.).

Author information

1
Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA; Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, D-01307 Dresden, Germany.

Abstract

Significant efforts have been made to identify HIV-1 neutralizing antibodies because they are considered to be critical to the design of an effective HIV-1 vaccine. Although soluble HIV-1 envelope proteins can be used for this purpose, these reagents differ from membrane-anchored HIV-1 envelope spike in a number of important ways and display only a subset of its native epitopes. Consistent with this, some broadly neutralizing antibodies preferentially bind cell surface-expressed HIV-1 envelope, but not the soluble protein. Here we report the details of a new method for isolating anti-HIV-1 specific B cells based on capturing cells that produce antibodies to cell surface-expressed gp160Δc(BaL). While this method is far less efficient than sorting with soluble envelope proteins, it isolated broadly neutralizing anti-HIV-1 antibodies that bind cell surface-expressed gp160Δc(BaL) but not soluble envelope proteins.

KEYWORDS:

HIV-1; HIV-1 neutralizing antibodies; Single B cell sort; Surface-expressed envelope trimer

PMID:
24041474
PMCID:
PMC3854001
DOI:
10.1016/j.jim.2013.09.003
[Indexed for MEDLINE]
Free PMC Article

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