Recombinant SARS-CoV-2 P.1 Spike GCN4-IZ Avi-tag Protein, CF

R&D Systems, part of Bio-Techne | Catalog # AVI10795

His-tag Biotinylated Gamma Variant (Brazil)
R&D Systems, part of Bio-Techne
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Key Product Details

Learn more about Avi-tag Biotinylated Proteins

Source

HEK293

Accession #

YP_009724390.1

Structure / Form

Biotinylated via Avi-tag

Conjugate

Biotin

Applications

Bioactivity

View all Spike Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein
SARS-CoV-2 P.1 Spike
(Val16-Lys1211)(Leu18Phe, Thr20Asn, Pro26Ser, Asp138Tyr, Arg190Ser, Lys417Thr, Glu484Lys, Asn501Tyr, Asp614Gly, His655Tyr, Thr1027Iso, Val1176Phe)(Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro)
Accession # YP_009724390.1		 GCN4-IZ Avi-tag 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

Val16

Predicted Molecular Mass

138 kDa

SDS-PAGE

140-170 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 P.1 Spike GCN4-IZ Avi-tag Protein, CF

Biotinylated Recombinant SARS-CoV-2 P.1 Spike (GCN4-IZ) Avi-tag His-tag Protein Binding Activity.

Biotinylated Recombinant SARS-CoV-2 P.1 Spike (GCN4-IZ) Avi-tag His-tag (Catalog # AVI10795) binds Recombinant Human ACE-2 Fc Chimera (10544-ZN) in a functional ELISA.

Biotinylated Recombinant SARS-CoV-2 P.1 Spike Avi-tag Protein SDS-PAGE.

2 μg/lane of Biotinylated Recombinant SARS-CoV-2 P.1 Spike Avi-tag Protein (Catalog # AVI10795) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 140-170 kDa.

Formulation, Preparation and Storage

AVI10795
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

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 SARS-CoV-2 shares 75% and 29% amino acid sequence identity with S protein of SARS-CoV-1 and MERS, respectively. 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). Several emerging SARS-CoV-2 genomes have been identified including the P.1 (Brazilian) variant (11). The P.1 variant contains 3 mutations of interest in the RBD domain: K417T, E484K, and N501Y (11). The E484K substitution alone has been shown to confer resistance to several monoclonal antibodies and is responsible for the first confirmed SARS-CoV-2 reinfection (12). Structural analysis points to E484K as a potentially crucial mutation as it creates a new site for hACE-2 binding and may enhance binding affinity (13). Our Avi-tag Biotinylated SARS-CoV-2 P.1 Spike (GCN4-IZ) features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of bionylation and the rest of the protein is unchanged so there is no interference in the protein bioactivity.

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, 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. https://doi.org/10.1016/j.it.2020.03.007.1.
  10. Okba, N.M.A. et al. (2020). Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
  11. Ferrareze, P.A.G. et al. (2021) bioRxiv https://doi.org/10.1101/2021.01.27.42689.
  12. Vasques Nonaka, C.K. et al. (2021) Emerg Infect Dis. https://doi.org/10.3201/eid2705.210191.
  13. Wang, W.B. et al. (2021) bioRxiv https://doi.org/10.1101/2021.02.17.431566.

Long Name

Spike Protein

Alternate Names

S Protein

Entrez Gene IDs

918758 (HCoV-229E); 2943499 (HCoV-NL63); 39105218 (HCoV-OC43); 37616432 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

YP_009724390.1

Additional Spike Products

Product Documents for Recombinant SARS-CoV-2 P.1 Spike GCN4-IZ Avi-tag Protein, CF

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Product Specific Notices for Recombinant SARS-CoV-2 P.1 Spike GCN4-IZ Avi-tag Protein, CF

For research use only

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