Monoclonal Antibodies to S and N SARS-CoV-2 Proteins as Probes to Assess Structural and Antigenic Properties of Coronaviruses.

Abstract

Antibodies targeting the spike (S) and nucleocapsid (N) proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are essential tools. In addition to important roles in the treatment and diagnosis of infection, the availability of high-quality specific antibodies for the S and N proteins is essential to facilitate basic research of virus replication and in the characterization of mutations responsible for variants of concern. We have developed panels of mouse and rabbit monoclonal antibodies (mAbs) to the SARS-CoV-2 spike receptor-binding domain (S-RBD) and N protein for functional and antigenic analyses. The mAbs to the S-RBD were tested for neutralization of native SARS-CoV-2, with several exhibiting neutralizing activity. The panels of mAbs to the N protein were assessed for cross-reactivity with the SARS-CoV and Middle East respiratory syndrome (MERS)-CoV N proteins and could be subdivided into sets that showed unique specificity for SARS-CoV-2 N protein, cross-reactivity between SARS-CoV-2 and SARS-CoV N proteins only, or cross-reactivity to all three coronavirus N proteins tested. Partial mapping of N-reactive mAbs were conducted using truncated fragments of the SARS-CoV-2 N protein and revealed near complete coverage of the N protein. Collectively, these sets of mouse and rabbit monoclonal antibodies can be used to examine structure/function studies for N proteins and to define the surface location of virus neutralizing epitopes on the RBD of the S protein.

• Authors: Rinki Kumar, Neil D Christensen, Rebecca J Kaddis Maldonado, Maria C Bewley, Alexandria Ostman, Malgorzata Sudol, Eunice C Chen, Natalie W Buchkovich, Abhinay Gontu, Meera Surendran Nair, Ruth H Nissly, Allen M Minns, Vivek Kapur, Randall Rossi, Suresh V Kuchipudi, Scott E Lindner, Leslie J Parent, John M Flanagan, Nicholas J Buchkovich
• Journal: Viruses (Viruses) Vol. 13 Issue 10 (09 22 2021) ISSN: 1999-4915 [Electronic] Switzerland
• PMID: 34696329 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Antibodies, Neutralizing
• Antibodies, Viral
• Coronavirus Nucleocapsid Proteins
• Epitopes
• Phosphoproteins
• Spike Glycoprotein, Coronavirus
• Viral Envelope Proteins
• nucleocapsid phosphoprotein, SARS-CoV-2
• spike protein, SARS-CoV-2

Topics

• Animals
• Antibodies, Monoclonal (immunology)
• Antibodies, Neutralizing (immunology)
• Antibodies, Viral (immunology)
• Betacoronavirus (immunology)
• Binding Sites (immunology)
• COVID-19 (immunology)
• Coronavirus Nucleocapsid Proteins (immunology, metabolism)
• Cross Reactions
• Epitopes (metabolism)
• Humans
• Mice
• Neutralization Tests
• Phosphoproteins (immunology, metabolism)
• Protein Binding (immunology)
• Rabbits
• SARS-CoV-2 (immunology, pathogenicity)
• Spike Glycoprotein, Coronavirus (genetics, immunology, metabolism)
• Viral Envelope Proteins (metabolism)