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

Huarui Qiao; Lingyun Li; Lu Wang; Haijun Yu; Fangzheng Hu; Xin Zhou; Hongzhao Yang; Jianfeng Xu; Xiangjing Meng; Yong Geng; Yuanyuan Dai

doi:10.1016/j.pep.2023.106267

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

Protein Expr Purif. 2023 Jul:207:106267. doi: 10.1016/j.pep.2023.106267. Epub 2023 Apr 6.

Authors

Huarui Qiao¹, Lingyun Li², Lu Wang³, Haijun Yu⁴, Fangzheng Hu¹, Xin Zhou¹, Hongzhao Yang¹, Jianfeng Xu⁵, Xiangjing Meng⁶, Yong Geng⁷, Yuanyuan Dai⁸

Affiliations

¹ Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China.
² The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
³ The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
⁴ State Key Laboratory of Drug Research & Center of Pharmaceutics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
⁵ Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China. Electronic address: jfxu@shou.edu.cn.
⁶ Shandong Provincial Key Laboratory of Biopharmaceuticals, Shandong Academy of Pharmaceutical Sciences, Jinan, 250101, China. Electronic address: fredamxj@163.com.
⁷ The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China; University of Chinese Academy of Sciences, Beijing, 100049, China. Electronic address: gengyong@simm.ac.cn.
⁸ Department of Pharmacy, National Cancer Center, National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China; National Cancer Center, National Clinical Research Center for Cancer/Hebei Cancer Hospital, Chinese Academy of Medical Sciences, Langfang, 065001, China. Electronic address: daiyuanyuan@cicams.ac.cn.

Abstract

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.

Keywords: Nanobody; PLpro inhibitor; Papain like protease (PLpro); SARS-CoV-2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19*
Coronavirus Papain-Like Proteases
Humans
Papain / chemistry
Peptide Hydrolases
SARS-CoV-2
Single-Domain Antibodies*

Substances

papain-like protease, SARS-CoV-2
Coronavirus Papain-Like Proteases
Single-Domain Antibodies
Peptide Hydrolases
Papain