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Here, we applied H2L2 transgenic mice encoding the human immunoglobulin variable regions, together with a state-of-the-art antibody discovery platform to immunize and isolate NAbs.
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Molecular understanding of neutralizing antibodies (NAbs) would greatly help advance the development of monoclonal antibody (mAb) therapy, as well as the design of next generation recombinant vaccines. The ongoing Coronavirus Disease 2019 (COVID-19) pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) threatens global public health and economy unprecedentedly, requiring accelerating development of prophylactic and therapeutic interventions. FOV, field of view NAb, neutralizing antibody SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus 2. (C) Representative fluorescent images of an FOV on the OptoSelect 14K chip and zoom in view of NanoPens with positive plasma B cells. The plasma B secreting anti-SARS-CoV-2 antibody was identified by increasing fluorescence near the opening of the NanoPen. (B) Bright field and fluorescent images of NanoPens with single plasma B cells. Antibody secreted by plasma B cells binding to antigen was detected by fluorescent anti-rat or anti-mouse IgG secondary antibody (green). Single plasma B cells (blue/purple) were loaded into individual NanoPens on the OptoSelect 14K chip in Beacon Optofluidic system, followed by import of a fluorescent secondary antibody-antigen beads mixture. Biotinylated antigen (dark blue) was conjugated to a streptavidin-decorated polystyrene bead (dark gray). (A) Schematic of in-chip assay on Beacon Optofluidic system to screening for anti-SARS-CoV-2 antibody secreting B cells.
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S1 Fig: Selection and characterization of NAbs.
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Dan Fu, Investigation, Methodology, Validation, # 1 Guangshun Zhang, Investigation, Methodology, Software, Validation, Visualization, # 1, 2, 3 Yuhui Wang, Investigation, Methodology, # 1, 3 Zheng Zhang, Investigation, Methodology, # 1 Hengrui Hu, Data curation, Formal analysis, Investigation, Methodology, # 4, 5 Shu Shen, Investigation, Methodology, # 4, 6 Jun Wu, Data curation, Investigation, Methodology, Software, Visualization, Writing – original draft, # 2 Bo Li, Investigation, Methodology, # 1, 3 Xin Li, Investigation, Methodology, Software, Visualization, # 1, 3, 7 Yaohui Fang, Investigation, Methodology, 6 Jia Liu, Investigation, Methodology, 4 Qiao Wang, Investigation, Methodology, Resources, Supervision, 8 Yunjiao Zhou, Investigation, Methodology, 8 Wei Wang, Investigation, Methodology, 7 Yufeng Li, Methodology, Project administration, 4 Zhonghua Lu, Investigation, Methodology, Software, 2 Xiaoxiao Wang, Investigation, Methodology, Supervision, 2 Cui Nie, Investigation, Methodology, 2 Yujie Tian, Investigation, Methodology, 1, 9 Da Chen, Methodology, Project administration, Software, Visualization, 1, 3, 9 Yuan Wang, Investigation, Methodology, Supervision, 1, 3, 9 Xingdong Zhou, Investigation, Methodology, 1, 3, 9 Qisheng Wang, Software, Supervision, Validation, Visualization, 10 Feng Yu, Investigation, Methodology, Software, 10 Chen Zhang, Investigation, Methodology, 1 Changjing Deng, Investigation, Methodology, 2 Liang Zhou, Investigation, Methodology, 2 Guangkuo Guan, Investigation, Methodology, 2 Na Shao, Investigation, Methodology, 2 Zhiyong Lou, Conceptualization, Project administration, Resources, 11, * Fei Deng, Investigation, Methodology, Project administration, Supervision, 4, 6, * Hongkai Zhang, Conceptualization, Funding acquisition, Supervision, Writing – original draft, 1, 7, 9, * Xinwen Chen, Conceptualization, Investigation, Resources, Writing – original draft, 5, 12, * Manli Wang, Investigation, Methodology, Supervision, Writing – review & editing, 4, * Louis Liu, Conceptualization, Funding acquisition, Investigation, Methodology, 2, * Zihe Rao, Conceptualization, Funding acquisition, Project administration, Supervision, Writing – original draft, 1, 3, 7, * and Yu Guo, Conceptualization, Formal analysis, Funding acquisition, Project administration, Supervision, Writing – original draft, Writing – review & editing 1, 9, 12, *