Mol Biomed. 2022 Jul 5
Meng Yu, Zhu Zhu, Yanqun Wang, Pingzhang Wang, Xiaodong Jia, Jie Wang, Lei Liu, Wanbing Liu, Yaqiong Zheng, Guomei Kou, Weiyan Xu, Jing Huang, Fengmin Lu, Xiajuan Zou, Shangen Zheng, Yinying Lu, Jincun Zhao, Hui Dai, and Xiaoyan Qiu
Products used in the paper Details Operation
hACE2-expressed HEK293 cell lines The pseudovirus neutralization assays were performed using hACE2-expressed HEK293 cell lines (purchase from PackGene Biotech). Request Quote

Research Field: Covid-related

Keywords: COVID-19, Neutralizing antibody, Neutralizing epitope;LC-MS/MS

AAV Serotype: hACE2-HEK293 cell lines


Although the SARS-CoV-2 vaccine has been widely used worldwide, not all individuals can produce neutralization antibodies, so it is still urgent to find and prepare neutralization antibodies for COVID-19 prevention or treatment. In this study, we created a new strategy to effectively obtain neutralizing antibodies or complementary determining region 3 (CDR3) of neutralizing antibodies against SARS-CoV-2. We first predicted and synthesized several B cell epitopes on RBD and adjacent RBD of S protein, then the B cell epitopes were used to prepare affinity chromatography columns respectively and purify the binding IgG from serum samples of convalescent COVID-19 patients. After these IgGs were identified to have neutralizing activity, the peptide sequences of the antigen-binding regions (variable region) of neutralizing antibodies were analyzed by protein mass spectrometry. Subsequently, the B cells from the same individual were sorted and used to obtain their full BCR repertoire by 5′ RACE combined with high-throughput of PacBio sequencing method. Then, the peptide sequence of neutralizing antibody variable region by protein mass spectrometry was mapped to the full BCR repertoire and found the full variable region sequence of neutralizing antibodies. Finally, we obtained and synthesized numerous CDR3 peptides of neutralizing antibodies to confirm the neutralizing activity for SARS-CoV-2 infection. Our results indicate that the novel scheme will be suitable for rapid screening of neutralizing antibodies, including screening neutralizing antibodies against SARS-CoV-2 and other pathogenic microorganisms.

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