Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry

STAR Protoc. 2021 Aug 24;2(3):100771. doi: 10.1016/j.xpro.2021.100771. eCollection 2021 Sep 17.


Protocols for efficient capture of antigen-specific B cells (ASBCs) are useful for understanding pathogen-specific B-cell responses during natural infection or vaccination. Fluorescently labeled tetramerized probes are classically used to capture ASBCs, but many occlude valuable epitopes available for B-cell receptor binding. Here, we describe a bead assay to confirm ASBC receptor accessibility on probes and a sequential staining process to capture HIV gp140-specific B cells from human peripheral blood mononuclear cells. For complete details on the use and execution of this protocol, please refer to Townsley et al. (2021).

Keywords: Flow Cytometry/Mass Cytometry; Immunology; Molecular/Chemical Probes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • B-Lymphocytes / immunology*
  • Flow Cytometry / methods*
  • Humans
  • Leukocytes, Mononuclear / immunology
  • Molecular Probes
  • Receptors, Antigen, B-Cell / metabolism
  • Staining and Labeling / methods*
  • env Gene Products, Human Immunodeficiency Virus* / chemistry


  • Molecular Probes
  • Receptors, Antigen, B-Cell
  • env Gene Products, Human Immunodeficiency Virus
  • gp140 envelope protein, Human immunodeficiency virus 1