ACE2 decoy receptor generated by high-throughput saturation mutagenesis efficiently neutralizes SARS-CoV-2 and its prevalent variants. Issue 1 (31st December 2022)

• Record Type: Journal Article
• Authors: Wang, Bolun; Zhao, Junxuan; Liu, Shuo; Feng, Jingyuan; Luo, Yufeng; He, Xinyu; Wang, Yanmin; Ge, Feixiang; Wang, Junyi; Ye, Buqing; Huang, Weijin; Bo, Xiaochen; Wang, Youchun; Xi, Jianzhong Jeff
• Is Part Of: Emerging microbes & infections. Volume 11:Issue 1(2022)
• Page Start: 1488
• Page End: 1499
• Publication Date: 2022-12-31
• Subjects: SARS-CoV-2 -- ACE2 -- high-throughput screening -- protein engineering -- decoy receptor; Medical microbiology -- Periodicals; Communicable diseases -- Periodicals; Infection -- Periodicals; 616.9041
• DOI: 10.1080/22221751.2022.2079426
• Languages: English
• ISSNs: 2222-1751

Abstract

ABSTRACT: The recent global pandemic was a spillover from the SARS-CoV-2 virus. Viral entry involves the receptor binding domain (RBD) of the viral spike protein interacting with the protease domain (PD) of the cellular receptor, ACE2. We hereby present a comprehensive mutational landscape of the effects of ACE2-PD point mutations on RBD-ACE2 binding using a saturation mutagenesis approach based on microarray-based oligo synthesis and a single-cell screening assay. We observed that changes in glycosylation sites and directly interacting sites of ACE2-PD significantly influenced ACE2-RBD binding. We further engineered an ACE2 decoy receptor with critical point mutations, D30I, L79W, T92N, N322V, and K475F, named C4-1. C4-1 shows a 200-fold increase in neutralization for the SARS-CoV-2 D614G pseudotyped virus compared to wild-type soluble ACE2 and a sevenfold increase in binding affinity to wild-type spike compared to the C-terminal Ig-Fc fused wild-type soluble ACE2. Moreover, C4-1 efficiently neutralized prevalent variants, especially the omicron variant (EC 50 = 16 ng/mL), and rescued monoclonal antibodies, vaccine, and convalescent sera neutralization from viral immune-escaping. We hope to next investigate translating the therapeutic potential of C4-1 for the treatment of SARS-CoV-2.