Assessment of Plasmodium falciparum Antigen-Specific B Cells

This protocol describes methods that exploit the specificity of binding between the B cell receptor and cognate antigen to detect and characterize Plasmodium-specific human B cells. Importantly, this approach allows for the isolation and study of B cells without activating the cells or requiring them to secrete antibodies. The protocol describes how antigen "probes" are generated and used in flow cytometry to identify and sort antigen-specific B cells, and includes methods for enrichment of antigen-specific B cells prior to flow cytometry. Finally, we detail techniques to optimize the exclusion of B cells that are not specific for the antigen of interest.