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

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

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Summary: 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 Passion Flower 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 Formal Shoes use and execution of this protocol, please refer to Townsley et al.(2021).