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

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


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / isolation & purification
  • Antibodies, Neutralizing / immunology
  • Antibodies, Neutralizing / isolation & purification
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism
  • Cell Line
  • HEK293 Cells
  • HIV Antibodies / immunology*
  • HIV Antibodies / isolation & purification*
  • HIV Envelope Protein gp160 / biosynthesis
  • HIV Envelope Protein gp160 / immunology*
  • HIV Envelope Protein gp160 / metabolism*
  • HIV-1 / immunology*
  • Humans


  • Antibodies, Monoclonal
  • Antibodies, Neutralizing
  • HIV Antibodies
  • HIV Envelope Protein gp160
  • gp160 protein, Human immunodeficiency virus 1