tetano
Editor, Senior Moderator
PLoS Pathog
. 2023 Feb 1;19(2):e1011119.
doi: 10.1371/journal.ppat.1011119. eCollection 2023 Feb.
High-throughput saturation mutagenesis generates a high-affinity antibody against SARS-CoV-2 variants using protein surface display assay on a human cell
Ye Yang[SUP] 1 [/SUP], Shuo Liu[SUP] 2 3 [/SUP], Yufeng Luo[SUP] 1 [/SUP], Bolun Wang[SUP] 1 [/SUP], Junyi Wang[SUP] 1 [/SUP], Juan Li[SUP] 1 [/SUP], Jiaxin Li[SUP] 1 [/SUP], Buqing Ye[SUP] 1 [/SUP], Youchun Wang[SUP] 3 [/SUP], Jianzhong Jeff Xi[SUP] 1 [/SUP]
Affiliations
Abstract
As new mutations continue to emerge, the ability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus to evade the human immune system and neutralizing antibodies remains a huge challenge for vaccine development and antibody research. The majority of neutralizing antibodies have reduced or lost activity against SARS-CoV-2 variants. In this study, we reported a novel protein surface display system on a mammalian cell for obtaining a higher-affinity antibody in high-throughput manner. Using a saturation mutagenesis strategy through integrating microarray-based oligonucleotide synthesis and single-cell screening assay, we generated a group of new antibodies against diverse prevalent SARS-CoV-2 variants through high-throughput screening the human antibody REGN10987 within 2 weeks. The affinity of those optimized antibodies to seven prevalent mutants was greatly improved, and the EC50 values were no higher than 5 ng/mL. These results demonstrate the robustness of our screening system in the rapid generation of an antibody with higher affinity against a new SARS-CoV-2 variant, and provides a potential application to other protein molecular interactions.
. 2023 Feb 1;19(2):e1011119.
doi: 10.1371/journal.ppat.1011119. eCollection 2023 Feb.
High-throughput saturation mutagenesis generates a high-affinity antibody against SARS-CoV-2 variants using protein surface display assay on a human cell
Ye Yang[SUP] 1 [/SUP], Shuo Liu[SUP] 2 3 [/SUP], Yufeng Luo[SUP] 1 [/SUP], Bolun Wang[SUP] 1 [/SUP], Junyi Wang[SUP] 1 [/SUP], Juan Li[SUP] 1 [/SUP], Jiaxin Li[SUP] 1 [/SUP], Buqing Ye[SUP] 1 [/SUP], Youchun Wang[SUP] 3 [/SUP], Jianzhong Jeff Xi[SUP] 1 [/SUP]
Affiliations
- PMID: 36724179
- DOI: 10.1371/journal.ppat.1011119
Abstract
As new mutations continue to emerge, the ability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus to evade the human immune system and neutralizing antibodies remains a huge challenge for vaccine development and antibody research. The majority of neutralizing antibodies have reduced or lost activity against SARS-CoV-2 variants. In this study, we reported a novel protein surface display system on a mammalian cell for obtaining a higher-affinity antibody in high-throughput manner. Using a saturation mutagenesis strategy through integrating microarray-based oligonucleotide synthesis and single-cell screening assay, we generated a group of new antibodies against diverse prevalent SARS-CoV-2 variants through high-throughput screening the human antibody REGN10987 within 2 weeks. The affinity of those optimized antibodies to seven prevalent mutants was greatly improved, and the EC50 values were no higher than 5 ng/mL. These results demonstrate the robustness of our screening system in the rapid generation of an antibody with higher affinity against a new SARS-CoV-2 variant, and provides a potential application to other protein molecular interactions.