Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD Fc Protein

Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD Fc Chimera (Catalog # 10717-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

2 μg/lane of Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD Fc Chimera (Catalog # 10717-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 55-65 kDa and 100-120 kDa, respectively.

Recombinant SARS-CoV-2 Spike RBD N501Y Fc was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.18 nM and 47.2 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=0.9196 nM.

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

Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD Fc Protein Summary

• Additional Information: N501Y Mutation found in UK, South African, and Brazilian Variants
• 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 RBD protein
  

  SARS-CoV-2 Spike RBD (Arg319-Phe541)(Asn501Tyr) Accession # YP_009724390.1	IEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: YP_009724390.1
• N-terminal Sequence: Arg319
• Structure / Form: Disulfide-linked homodimer
• 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: 52 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: 55-65 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 B.1.1.7 N501Y Spike RBD Fc Protein

• Spike RBD

Background

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that also include MERS and SARS-CoV-1. Coronaviruses are commonly comprised of four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M) and Nucleocapsid protein (N) (1). The SARS-CoV-2 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 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). A metallopeptidase, angiotensin-converting enzyme 2 (ACE2), has been identified as a functional receptor for SARS-CoV-2 through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (6,7). The RBD of SARS-CoV-2 shares 73% aa identity with the RBD of the SARS-CoV-1, but only 22% amino acid (aa) identity with the RBD of MERS. A SARS-CoV-2 variant carrying the aa substitution N501Y in the RBD is one of the most prevalent mutations found Covid-19 cases (8-10). This mutation was first identified in the virus variant (B.1.1.7 lineage) originally found in London and the southeast UK but rapidly spread globally (8,9). This new virus variant was reported 56% more transmissible than other preexisting variants (11). The N501Y mutation was also later identified in variants found in South Arica (B.1.351 lineage) and Brazil (P.1 lineage). Although there is no evidence to date that B.1.1.7 causes more severe illness, whether the N501Y mutation in RBD would decrease the efficacy of vaccine-induced immunity is still under investigation.

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. Li, W. et al. (2003) Nature 426:450.
7. Wong, S. K. et al. (2004) J. Biol. Chem. 279:3197.
8. Kozlov, Max (2020) TheScientist https://www.the-scientist.com/news-opinion/new-sars-cov-2-variant-spreading-rapidly-in-uk-68292.
9. Wise, J. (2020) B.M.J. 371:m4857.
10. Tang, J.W. et al. (2020) J. Infect. doi: 10.1016/j.jinf.2020.12.024.
11. Davies, N.G. (2020) medRxiv doi:10.1101/2020.12.24.20248822.

Bioinformatics

• Uniprot:
  • YP_009724390.1(Rabbit)