This protocol is a comprehensive guide to phage display-based selection of virus neutralizing V antibody domains. It details three optimized parts including (1) construction of a large-sized (theoretically > 10) 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 V antibody domain, V ab8, which exhibits high prophylactic and therapeutic efficacy. For complete details on the use and execution of this protocol, please refer to Li et al. (2020).