Preparation and characterization of nanobodies targeting SARS-CoV-2 papain-like protease

Coronavirus Papain-like protease (PLpro) mediates the cleavage of viral polyproteins and assists the virus escaping from innate immune response. Thus, PLpro is an attractive target for the development of broad-spectrum drugs as it has a conserved structure across different coronaviruses. In this study, we purified SARS-CoV-2 PLpro as an immune antigen, constructed a nanobody phage display library, and identified a set of nanobodies with high affinity for SARS-CoV-2. In addition, enzyme activity experiments demonstrated that two nanobodies had a significant inhibitory effect on the PLpro. These nanobodies should therefore be investigated as candidates for the treatment of coronaviruses. •A nanobody phage library of SARS-CoV-2 papain-like protease (PLpro)was successfully constructed.•The nanobodies were successfully purified and demonstrated to have high affinity for SARS-CoV-2 PLpro.•The nanobodies have a significant inhibitory effect on SARS-CoV-2 PLpro.