Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF

Recombinant SARS-CoV-2 Spike S1 Subunit His-tag (Catalog # 10569-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

2 μg/lane of Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF (Catalog # 10569-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 106-121 kDa.

Recombinant SARS-CoV-2 Spike S1 Subunit His tag protein (10569-CV) was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.184 nM and 93.5 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=2.625 nM.

• Reactivity: V
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF Summary

• Additional Information: HEK293 Expressed
• Details of Functionality: Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag (Catalog # 933-ZN).
• Source: Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike S1 Subunit protein
  Val16-Pro681, with a C-terminal 6-His tag
• Accession #: YP_009724390.1
• N-terminal Sequence: Val16
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 75 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 106-121 kDa, under reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF

• SARS-CoV-2
• Spike S1 Subunit

Background

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that are commonly comprised of four structural proteins: Spike protein(S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). SARS-CoV-2 Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry. The S protein is homotrimeric, with each ~180-kDa monomer consisting of two subunits, S1 and S2 (2). In SARS-CoV-2, as with most coronaviruses, proteolytic cleavage of the S protein into two distinct peptides, S1 and S2 subunits, is required for activation. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion (3-5). Based on structural biology studies, the receptor binding domain (RBD), located in the C-terminal region of S1, can be oriented either in the up/standing or down/lying state (6). The standing state is associated with higher pathogenicity and both SARS-CoV-1 and MERS can access this state due to the flexibility in their respective RBDs. A similar two-state structure and flexibility is found in the SARS-CoV-2 RBD (7). Based on amino acid (aa) sequence homology, the SARS-CoV-2 S1 subunit has 65% identity with SARS-CoV-1 S1 subunit, but only 22% homology with the MERS S1 subunit. The low aa sequence homology is consistent with the finding that SARS and MERS bind different cellular receptors (8). The S Protein of the SARS-CoV-2 virus, like the SARS-CoV-1 counterpart, binds Angiotensin-Converting Enzyme 2 (ACE-2), but with much higher affinity and faster binding kinetics (9). Before binding to the ACE-2 receptor, structural analysis of the S1 trimer shows that only one of the three RBD domains in the trimeric structure is in the "up" conformation. This is an unstable and transient state that passes between trimeric subunits but is nevertheless an exposed state to be targeted for neutralizing antibody therapy (10). Polyclonal antibodies to the RBD of the SARS-CoV-2 S1 subunit have been shown to inhibit interaction with the ACE-2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (11). There is also promising work showing that the RBD may be used to detect presence of neutralizing antibodies present in a patient's bloodstream, consistent with developed immunity after exposure to the SARS-CoV-2 virus (12). Lastly, it has been demonstrated the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (13, 14).

1. Wu, F. et al. (2020) Nature 579:265.
2. Tortorici, M.A. and D. Veesler (2019). Adv. Virus Res. 105:93.
3. Bosch, B.J. et al. (2003). J. Virol. 77:8801.
4. Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. 106:5871.
5. Millet, J.K. and G. R. Whittaker (2015) Virus Res. 202:120.
6. Yuan, Y. et al. (2017) Nat. Commun. 8:15092.
7. Walls, A.C. et al. (2010) Cell 180:281.
8. Jiang, S. et al. (2020) Trends. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
9. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
10. Wrapp, D. et al. (2020) Science 367:1260.
11. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
12. Okba, N. M. A. et al. (2020). Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
13. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
14. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.

Publications for SARS-CoV-2 Spike S1 Protein (10569-CV)(6)

Publications using 10569-CV

• Haystead, T;Lee, E;Cho, K;Gullickson, G;Hughes, P;Krafsur, G;Freeze, R;Scarneo, S; Investigation of SARS-CoV-2 individual proteins reveals the in vitro and in vivo immunogenicity of membrane protein Scientific reports 2023-12-18 [PMID: 38129491] (Bioassay, Human)
• Kiszel, P;Sík, P;Miklós, J;Kajdácsi, E;Sinkovits, G;Cervenak, L;Prohászka, Z; Class switch towards spike protein-specific IgG4 antibodies after SARS-CoV-2 mRNA vaccination depends on prior infection history Scientific reports 2023-08-13 [PMID: 37574522] (ELISA Capture, Human)
• ML Berre, T Paulov?áko, CM Verissimo, S Doyle, JP Dalton, C Masterson, ER Martínez, L Walsh, C Gormley, JG Laffey, B McNicholas, AJ Simpkin, M Kilcoyne A new multiplex SARS-CoV-2 antigen microarray showed correlation of IgG, IgA, and IgM antibodies from patients with COVID-19 disease severity and maintenance of relative IgA and IgM antigen binding over time PLoS ONE, 2023;18(3):e0283537. 2023 [PMID: 36996259] (Binding, Human)
• ML Berre, T Paulov?áko, CM Verissimo, S Doyle, JP Dalton, C Masterson, ER Martínez, L Walsh, C Gormley, JG Laffey, B McNicholas, AJ Simpkin, M Kilcoyne A new multiplex SARS-CoV-2 antigen microarray showed correlation of IgG, IgA, and IgM antibodies from patients with COVID-19 disease severity and maintenance of relative IgA and IgM antigen binding over time PLoS ONE, 2023-03-30;18(3):e0283537. 2023-03-30 [PMID: 36996259] (Binding, Human)
• G Sinkovits, J Schnur, L Hurler, P Kiszel, ZZ Prohászka, P Sík, E Kajdácsi, L Cervenak, V Maráczi, M Dávid, B Zsigmond, É Rimanóczy, C Bereczki, L Willems, EJM Toonen, Z Prohászka Evidence, detailed characterization and clinical context of complement activation in acute multisystem inflammatory syndrome in children Scientific Reports, 2022-11-17;12(1):19759. 2022-11-17 [PMID: 36396679] (ELISA Standard, Human)
• AC Dowell, MS Butler, E Jinks, G Tut, T Lancaster, P Sylla, J Begum, R Bruton, H Pearce, K Verma, N Logan, G Tyson, E Spalkova, S Margielews, GS Taylor, E Syrimi, F Baawuah, J Beckmann, IO Okike, S Ahmad, J Garstang, AJ Brent, B Brent, G Ireland, F Aiano, Z Amin-Chowd, S Jones, R Borrow, E Linley, J Wright, R Azad, D Waiblinger, C Davis, EC Thomson, M Palmarini, BJ Willett, WS Barclay, J Poh, G Amirthalin, KE Brown, ME Ramsay, J Zuo, P Moss, S Ladhani Children develop robust and sustained cross-reactive spike-specific immune responses to SARS-CoV-2 infection Nature Immunology, 2021-12-22;23(1):40-49. 2021-12-22 [PMID: 34937928] (Neutralization, Human)

Bioinformatics

• Uniprot:
  • YP_009724390.1(Llama)