Recombinant SARS-CoV-2 B.1.351 Spike GCN4-IZ His Protein, CF
Recombinant SARS-CoV-2 B.1.351 Spike GCN4-IZ His Protein, CF Summary
Product Specifications
| SARS-CoV-2 Spike (Val16-Lys1211)(Asp80Ala, Asp215Gly, Leu242del, Ala243del, Leu244del, Lys417Asn, Glu484Lys, Asn501Tyr, Asp614Gly, Ala701Val)(Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro) Accession # YP_009724390.1 | GCN4-IZ | 6-His tag |
| N-terminus | C-terminus | |
Analysis
Product Datasheets
10786-CV
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
10786-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.
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Scientific Data
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Recombinant SARS-CoV-2 B.1.351 Spike (GCN4-IZ) His-tag (Catalog # 10786-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.
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2 μg/lane of Recombinant SARS-CoV-2 B.1.351 Spike GCN4-IZ His-tag (Catalog # 10786-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 145-168 kDa.
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Recombinant SARS-CoV-2-B.1.351 South Africa variant Spike protein was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.046 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=1.272 nM.
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 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 the 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 (aa) sequence identity with the 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 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 (6). Based on structural biology studies, the RBD can be oriented either in the up/standing or down/lying state with the up/standing state associated with higher pathogenicity (7). 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 (8). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (9, 10). A SARS-CoV-2 variant carrying amino acid substitutions N501Y, K417N, and E484K in the RBD raised the most concerns. This B.1.351 lineage, also known and 501Y.V2 variant, was first identified in the Eastern Cape province of South Africa in December 2020 and spread quickly to become the most dominant strain in the second COVID wave in South Africa (11). Two of these mutations K417N and E484K locate at the receptor binding motif (RBM) and are not found in other variants (11). The N501Y mutation is also found in London (B.1.1.7 lineage) and Brazil (P.1 lineage). The B.1.351 lineage is reported to enter cells more easily due to its enhanced affinity to ACE-2 receptor (12). It is reported to reduce the efficacy of neutralizing antibody (12, 13).
- Wu, F. et al. (2020) Nature 579:265.
- Tortorici, M.A. and D. Veesler (2019) Adv. Virus Res. 105:93.
- Bosch, B.J. et al. (2003) J. Virol. 77:8801.
- Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. 106:5871.
- Millet, J.K. and G.R. Whittaker (2015) Virus Res. 202:120.
- Ortega, J.T. et al. (2020) EXCLI J. 19:410.
- Yuan, Y. et al. (2017) Nat. Commun. 8:15092.
- Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
- Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
- Wang, K. et al. (2020) bioRxiv https://doi.org/10.1101/2020.03.14.988345.
- Tegally, H. et al. (2020) bioRxiv. Doi: https://doi.org/10.1101/2020.12.21.20248640.
- Nelson, G. et al. (2021) bioRxiv. https://doi: 10.1101/2021.01.13.426558.
- Wibmer, C.K. et al. (2021) bioRxiv. https://doi: 10.1101/2021.01.18.427166.
Citations for Recombinant SARS-CoV-2 B.1.351 Spike GCN4-IZ His Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Site specific N- and O-glycosylation mapping of the spike proteins of SARS-CoV-2 variants of concern
Authors: Shajahan, A;Pepi, LE;Kumar, B;Murray, NB;Azadi, P;
Scientific reports
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Site Specific N- and O-glycosylation mapping of the Spike Proteins of SARS-CoV-2 Variants of Concern
Authors: A Shajahan, L Pepi, B Kumar, N Murray, P Azadi
Research square, 2022-11-16;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
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