B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 Receptor Binding Domain

ABSTRACT

B-cell ELISpot is an extremely sensitive assay based on the secretion of antibodies by B cells which requires the differentiation of B cells into antibody-secreting cells (ASCs). Here, we describe the procedure to analyse both plasmablast (PB) and memory B cell (MBC) responses specific to SARS-CoV-2 Receptor Binding Domain in the context of acute SARS-CoV-2 infection and vaccination. We detail steps for MBC stimulation, MBC and PB plating, detection and counting of total IgG and RBD-specific spots. Graphical Publisher's note Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.