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

Q2 Medicine Antibody Therapeutics Pub Date : 2023-04-01 DOI:10.1093/abt/tbad002

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

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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.

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利用高通量单b细胞克隆方法制备抗SARS-CoV-2的中和抗体。

背景:需要快速有效的策略从病毒感染患者的B细胞中发现中和抗体(nab)。方法:本文报道了一种高通量单b细胞克隆方法，用于高通量分离针对COVID-19恢复期患者SARS-CoV-2-RBD(受体结合域)不同表位的nab。该方法简单、快速、高效地从COVID-19患者的B细胞中生成sars - cov -2中和抗体。结果:利用该方法，我们开发了针对不同SARS-CoV-2-RBD表位的多个nab。低温电镜和晶体学精确地揭示了它们是如何结合RBD的。在活病毒实验中，这些nab能有效阻断病毒进入宿主细胞。结论:该方法简便有效，可用于其它疾病和下一次大流行的人类治疗性抗体的研制。

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