Protocol to isolate broadly neutralizing monoclonal antibodies against SARS-CoV-2 from human B cells
Broadly neutralizing monoclonal antibodies (bnAbs) against SARS-CoV-2 are crucial for developing therapeutics and vaccines that confer broad protection against SARS-CoV-2 variants and related sarbecoviruses. Here, we present a protocol to isolate bnAbs against SARS-CoV-2 from human B cells. We describe steps for sorting single spike-specific B cells from human peripheral blood mononuclear cells, cloning monoclonal antibodies (mAbs) from single B cells, screening mAbs with broadly neutralizing activity, and measuring the neutralizing potency of bnAbs against a panel of pseudoviruses. For complete details on the use and execution of this protocol, please refer to Chen et al.<sup>1</sup>.
Authors
- Ruiping Du
- Xueyan Huang (ORCID: https://orcid.org/0000-0002-5831-1456)
- Yunjian Li (ORCID: https://orcid.org/0000-0002-5017-8996)
- Liwei Jiang (ORCID: https://orcid.org/0000-0001-5990-2509)
- Xixian Chen
- Yi Sun (ORCID: https://orcid.org/0000-0003-0474-822X)
- Ling Li (ORCID: https://orcid.org/0000-0002-2763-0761)
- Yu-Cong Sun
- Maosheng Lu
- Teng Zuo (ORCID: https://orcid.org/0009-0002-7399-3229)
- Zuowei Wang (ORCID: https://orcid.org/0000-0002-1929-231X)
- Lin Hu
- Rongrong Qin
Institutions
- Chinese Academy of Sciences (CN)
- Hefei Institutes of Physical Science (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-12-12
- DOI
- https://doi.org/10.1016/j.xpro.2025.104275
- Primary Topic
- SARS-CoV-2 and COVID-19 Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00