Development of neutralizing antibodies against SARS-CoV-2, using a high-throughput single-B-cell cloning method

Yang Dou; Ke Xu; Yong-Qiang Deng; Zijing Jia; Jun Lan; Xiaoyu Xu; Guorui Zhang; Tianshu Cao; Pan Liu; Xiangxi Wang; Xinquan Wang; Lingjie Xu; Pan Du; Cheng-Feng Qin; Hong Liu; Yafeng Li; Guizhen Wu; Kang Wang; Bai Lu

doi:10.1093/abt/tbad002

Development of neutralizing antibodies against SARS-CoV-2, using a high-throughput single-B-cell cloning method

Antib Ther. 2023 Feb 17;6(2):76-86. doi: 10.1093/abt/tbad002. eCollection 2023 Apr.

Authors

Yang Dou¹, Ke Xu², Yong-Qiang Deng³, Zijing Jia⁴, Jun Lan⁵, Xiaoyu Xu⁶, Guorui Zhang⁷, Tianshu Cao⁴, Pan Liu⁴, Xiangxi Wang⁴, Xinquan Wang⁵, Lingjie Xu⁶, Pan Du⁶, Cheng-Feng Qin³, Hong Liu⁷, Yafeng Li⁸, Guizhen Wu², Kang Wang⁴, Bai Lu¹

Affiliations

¹ School of Pharmaceutical Sciences, IDG/McGovern Institute for Brain Research, Tsinghua University, Beijing 100084, China.
² NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
³ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, AMMS, Beijing 100071, China.
⁴ National Laboratory of Macromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
⁵ Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
⁶ Vazyme Biotech Co., Ltd., Nanjing, China.
⁷ Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
⁸ Shanxi Provincial Key Laboratory of Kidney Disease, The Fifth Hospital of Shanxi Medical University, Taiyuan 030012, China.

Abstract

Background: Rapid and efficient strategies are needed to discover neutralizing antibodies (nAbs) from B cells derived from virus-infected patients.

Methods: Here, we report a high-throughput single-B-cell cloning method for high-throughput isolation of nAbs targeting diverse epitopes on the SARS-CoV-2-RBD (receptor binding domain) from convalescent COVID-19 patients. This method is simple, fast and highly efficient in generating SARS-CoV-2-neutralizing antibodies from COVID-19 patients' B cells.

Results: Using this method, we have developed multiple nAbs against distinct SARS-CoV-2-RBD epitopes. CryoEM and crystallography revealed precisely how they bind RBD. In live virus assay, these nAbs are effective in blocking viral entry to the host cells.

Conclusion: This simple and efficient method may be useful in developing human therapeutic antibodies for other diseases and next pandemic.