title: Isolation of HIV-1-reactive antibodies using cell surface-expressed gp160Δc(BaL.).
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.
authors.affiliation: 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.
authors.forename: Christian
authors.initials: C
authors.lastname: Gaebler
authors.forename: Henning
authors.initials: H
authors.lastname: Gruell
authors.forename: Klara
authors.initials: K
authors.lastname: Velinzon
authors.forename: Johannes F
authors.initials: JF
authors.lastname: Scheid
authors.forename: Michel C
authors.initials: MC
authors.lastname: Nussenzweig
authors.forename: Florian
authors.initials: F
authors.lastname: Klein
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000276:immunology / Q000302:isolation & purification; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D057809:HEK293 Cells; D015483:HIV Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D019302:HIV Envelope Protein gp160 / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D015483:HIV Antibodies; D019302:HIV Envelope Protein gp160; C514797:gp160 protein, Human immunodeficiency virus 1
keywords: HIV-1; HIV-1 neutralizing antibodies; Single B cell sort; Surface-expressed envelope trimer
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