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Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 11442-CV

R&D Systems, part of Bio-Techne
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11442-CV-100

Key Product Details

Source

HEK293

Accession #

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike RBD protein
SARS-CoV-2 EG.5.1 Spike RBD
(Arg319-Phe541)
(Gly339His, Arg346Thr, Leu368Ile, Ser371Phe, Ser373Pro, Ser375Phe, Thr376Ala, Asp405Asn, Arg408Ser, Lys417Asn, Asn440Lys, Val445Pro, Gly446Ser, Phe456Leu, Asn460Lys, Ser477Asn, Thr478Lys, Glu484Ala, Phe486Pro, Phe490Ser, Gln498Arg, Asn501Tyr, Tyr505His)
Accession # YP_009724390.1
6-His tag
N-terminus C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Arg319

Predicted Molecular Mass

26 kDa

SDS-PAGE

34-38 kDa, under reducing conditions.

Activity

Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 Fc Chimera (Catalog # 10544-ZN).

Scientific Data Images for Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein, CF

Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein Binding Activity.

Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein (Catalog # 11442-CV) binds Recombinant Human ACE-2 Fc Chimera (Catalog # 10544-ZN) in a functional ELISA.

Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein (Catalog # 11442-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 34-38 kDa, under reducing conditions.

Formulation, Preparation and Storage

11442-CV
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Spike RBD

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-CoV 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).  The S Protein of the SARS-CoV-2 virus, like the SARS-CoV-1 counterpart, binds a metallopeptidase, Angiotensin-Converting Enzyme 2 (ACE-2), but with much higher affinity and faster binding kinetics through the receptor binding domain (RBD) located in the C-terminal region of S1 subunit (6). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (7, 8). Polyclonal antibodies to the RBD of the SARS-CoV-2 protein have been shown to inhibit interaction with the ACE-2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (9). 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 (10). Based on amino acid (aa) sequence homology, the SARS-CoV-2 RBD has 73% identity with SARS-CoV-1 RBD, but only 22% homology with the MERS RBD. Several emerging SARS-CoV-2 genomes have been identified including the Omicron, or B.1.1.529, variant. First identified in November 2021 in South Africa, the Omicron variant quickly became the predominant SARS-CoV-2 variant and is considered a variant of concern (VOC). The Omicron variant contains 32 mutations in the S protein, 3 to 4 times more than in other SARS-CoV-2 variants, that potentially affect viral fitness and transmissibility (11). Of these mutations,15 are located in the RBD domain and allow the Omicron variant to bind ACE-2 with greater affinity and, potentially, increased transmissibility (11, 12). Several additional mutations throughout the S protein have been shown or are predicted to enhance spike cleavage and could aid transmission (13-15). The study of the Omicron variant's impact on immune escape and reduced neutralization activity to monoclonal antibodies along with an increased risk of reinfection, even among vaccinated individuals, remains ongoing (16). The EG.5.1 variant was evolved from Omicron subvariant XBB.1.9 with an additional F456L substitiution within RBD (17).

References

  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. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  7. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.
  8. Isabel, S. et al. (2020) Sci Rep. 10, 14031. https://doi.org/10.1038/s41598-020-70827-z.
  9. Tai, W. et al. (2020) Cell. Mol. Immunol. 17:613.
  10. Okba, N. M. A. et al. (2020). Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
  11. Shah, M. and Woo, H.G. (2021) bioRxiv https://doi.org/10.1101/2021.12.04.471200.
  12. Lupala, C.S. et al. (2021) bioRxiv https://doi.org/10.1101/2021.12.10.472102.
  13. Zhang, L. et al. (2020) Nat Commun. 11:6013.
  14. Lasek-Nesselquist, E. et al. (2021) medRxiv https://doi.org/10.1101/2021.03.10.21253285.
  15. Scheepers, C. et al. (2021) medRxiv https://doi.org/10.1101/2021.08.20.21262342.
  16. Callaway, E. and Ledford, H. (2021) Nature 600:197.
  17. Wang, Q. et al. (2023)  bioRxiv https://doi.org/10.1101/2023.08.21.553968.

Long Name

Spike Receptor Binding Domain

Entrez Gene IDs

3200426 (HCoV-HKU1); 14254594 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

Additional Spike RBD Products

Product Documents for Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant SARS-CoV-2 EG.5.1 Spike RBD His-tag Protein, CF

For research use only

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