This protocol is a comprehensive guide to phage display based selection of virus neutralizing VH antibody domains. It details three optimized parts including 1) Construction of large size (theoretically > 1011) naïve human antibody heavy chain domain library, 2) SARS-COV-2 antigen expression and stable cell line construction, and 3) Library panning for selection of SARS-CoV-2 specific antibody domains. Using this protocol, we identified a high-affinity neutralizing human VH antibody domain, VH ab8, which exhibits high prophylactic and therapeutic efficacy.
Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains