SARS-CoV-2 Spike Antibody (CR3022)

ELISA: SARS-CoV-2 Spike Antibody (CR3022) [NBP2-90980] - Binding curve of SARS-CoV-2 Spike Antibody (CR3022) to SARS-CoV-2 Spike Glycoprotein domains S1 and S2 of various origin. ELISA plate coated with SARS-CoV-2 Spike Glycoprotein (S1), His-Tag (Insect Cells; grey line), SARS-CoV-2 Spike Glycoprotein (S2), His-Tag (Insect Cells; yellow line) and SARS Coronavirus Spike Glycoprotein (S1), His-Tag (HEK293 cells; blue line) (Native Antigen) at concentrations of 5 ug/ml. A 3-fold serial dilution from 41.6 ng/ml was performed using SARS-CoV-2 Spike Antibody (CR3022). For detection, a 1:4000 dilution of HRP-labelled anti-human IgG antibody was used.

ELISA: SARS-CoV-2 Spike Antibody (CR3022) [NBP2-90980] - Binding curve of SARS-CoV-2 Spike Antibody (CR3022) to SARS-CoV-2 Spike Glycoprotein (S1), Sheep Fc-Tag and SARS-CoV-2 Spike Glycoprotein (S2), Sheep Fc-Tag from HEK293 cells. ELISA plate coated with SARS-CoV-2 Spike Glycoprotein (S1), Sheep Fc-Tag (blue line) or SARS-CoV-2 Spike Glycoprotein (S2), Sheep Fc-Tag (orange line) from HEK293 cells (Native Antigen) at concentrations of 5 ug/ml. A 3-fold serial dilution from 125 ng/ml was performed using SARS-CoV-2 Spike Antibody (CR3022). For detection, a 1:4000 dilution of HRP-labelled anti-human IgG antibody was used.

• Reactivity: V
• Applications: ELISA, ICC/IF, SPR
• Clone: CR3022
• Clonality: Monoclonal
• Host: Human
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

SARS-CoV-2 Spike Antibody (CR3022) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: The original monoclonal antibody was generated by sequencing peripheral blood lymphocytes of a patient exposed to the SARS-CoV.
• Specificity: This antibody binds to both SARS-CoV and SARS-CoV-2 with high affinity (PMID: 16796401 & 32065055). It binds the amino acids 318-510 in the S1 domain of the SARS-CoV Spike protein as well as SARS-CoV-2 (COVID-19) Spike protein. The antibody also binds to P462L-substituted S318-510 fragments of the SARS spike protein. The binding epitope is only accessible in the "open" confromation of the spike protein (Joyce et al. 2020).
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Human
• Gene: S
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunocytochemistry/Immunofluorescence
  • Surface Plasmon Resonance
• Application Notes: The initial characterization of the binding of this antibody was performed by ELISA and indicates potential for the development of diagnostic assays, as both virus-capture assays, or as controls in serological assays measuring immune-responses to virus exposure. Human IgG1, IgG3, IgM and IgA isotypes are available to mimic antibody responses seen in COVID19 (Amanat et al. 2020). Human IgG2 is available to assess its yet unknown role in the response to SARS-CoV-2. The original human IgG1 version of the antibody works synergistically in combination with another non-competing SARS antibody and is a potential candidate for passive immune prophylaxis of SARS-CoV infection (Meulen et al., 2006). The original antibody (human IgG1) was also reported to bind the 2019-nCoV RBD (KD of 6.3 nM). This antibody has been attributed a potential to be developed as a therapeutic agent, alone or in combination with other neutralizing antibodies for treatment of 2019-nCoV infections (Tian et al., 2020).

Reactivity Notes

SARS-CoV, SARS-CoV-2

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Proclin 300
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for SARS-CoV-2 Spike Antibody (CR3022)

• 2019-nCoV S Protein
• 2019-nCoV Spike
• COVID-19 Spike
• E2
• Human coronavirus spike glycoprotein
• Peplomer protein
• S glycoprotein
• S Protein
• SARS-COV-2 S protein
• SARS-COV-2 Spike glycoprotein
• SARSCOV2 Spike protein
• SARS-CoV-2
• Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein
• Spike glycoprotein
• Spike
• surface glycoprotein

Background

The SARS-CoV-2 Spike protein is one of the four major structural proteins of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of COVID-19 (1,2). The spike protein is the largest of the structural proteins, which also include the membrane (M), envelope (E), and nucleocapsid (N) proteins (1,2). The SARS-CoV-2 spike protein is a 1273 amino acid (aa) heterotrimeric class I fusion protein with each monomer having a theoretical molecular weight of approximately 180 kDa (1). The club-shaped spike protein contains several functional regions and domains including the S1 globular head region which contains the N-terminal receptor-binding domain (RBD) and the S2 stem region that contains the C-terminal fusion domain, two heptad regions, a transmembrane domain, and a cytoplasmic tail (1,2). The viral spike protein is critical for attachment of the virus with the host cell, resulting in fusion and virus entry into the cell (1,2). More specifically, the RBD of the spike protein is responsible for binding to the cell surface receptor angiotensin converting enzyme 2 (ACE2) (1,2). This spike-ACE2 interaction results in a conformational change permitting furin cleavage between the S1 and S2 domains and then cleavage at S2' by TMPRRS2, or another protease, allowing membrane fusion (1,2).

Given the critical role of the spike protein RBD in the interaction with the ACE2 receptor and viral entry, a number of neutralizing antibodies against the RBD have been developed as potential therapeutics for treating COVID-19 (3). These antibodies bind the RBD domain on the S1 subunit inhibiting the interaction with ACE2 (3). However, more studies need to be done as neutralizing antibodies can result in antibody-dependent enhancement, in which the viral entry and replication within the host cell is increased (4). One potential way to combat antibody-dependent enhancement is the use of nanobodies (4). Furthermore, there are currently several vaccine strategies that are in clinical trials, or recently federally approved, that utilize the spike protein in different forms (e.g. full length, S1 RBD, RBD-Fc, N-terminal) for protecting against SARS-CoV-2 infection (4,5). These vaccine strategies include DNA vaccines, viral vector-based vaccines, RNA vaccines, and subunit vaccines (4,5).

References

1. Pillay T. S. (2020). Gene of the month: the 2019-nCoV/SARS-CoV-2 novel coronavirus spike protein. Journal of Clinical Pathology. https://doi.org/10.1136/jclinpath-2020-206658

2. Malik Y. A. (2020). Properties of Coronavirus and SARS-CoV-2. The Malaysian Journal of Pathology.

3. Ho M. (2020). Perspectives on the development of neutralizing antibodies against SARS-CoV-2. Antibody Therapeutics. https://doi.org/10.1093/abt/tbaa009

4. Samrat, S. K., Tharappel, A. M., Li, Z., & Li, H. (2020). Prospect of SARS-CoV-2 spike protein: Potential role in vaccine and therapeutic development. Virus Research. https://doi.org/10.1016/j.virusres.2020.198141

5. Sternberg, A., & Naujokat, C. (2020). Structural features of coronavirus SARS-CoV-2 spike protein: Targets for vaccination. Life Sciences. https://doi.org/10.1016/j.lfs.2020.118056

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SARS-CoV-2 Spike Antibody (NBP2-90980)(4)

Publications using NBP2-90980

• Kaneko T, Esmail S, Voss C et al. System-wide hematopoietic and immune signaling aberrations in COVID-19 revealed by deep proteome and phosphoproteome analysis Research Square Feb 10 2021 (MiAr)
• Esmail S, Knauer M, Abdoh H et al. Rapid and accurate agglutination-based testing for SARS-CoV-2 antibodies Cell Reports Methods May 1 2021
• Staufer O, Gupta K, Hernandez BUcher JE et al. Synthetic virions reveal fatty acid-coupled adaptive immunogenicity of SARS-CoV-2 spike glycoprotein Nature communications Feb 14 2022 [PMID: 35165285] (SARS-CoV-2)
• Cai Q, Mu J, Lei Y et al. Simultaneous detection of the spike and nucleocapsid proteins from SARS-CoV-2 based on ultrasensitive single molecule assays Anal Bioanal Chem May 31 2021 [PMID: 34057558]

Bioinformatics

• Gene Symbol: S
• Uniprot:
  • P59594(Virus)