Recombinant SARS-CoV-2 Y453F Spike RBD Fc Chimera Protein CF

Name: Recombinant SARS-CoV-2 Y453F Spike RBD Fc Chimera Protein CF
Brand: R&D Systems
SKU: 10792-CV

Catalog # 10792-CV

Product Datasheet / COA / SDS

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Key Product Details

Source:	HEK293
Applications:	Bioactivity

View Full Technical Details for Recombinant SARS-CoV-2 Y453F Spike RBD Fc Chimera Protein CF

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike RBD protein

SARS-CoV-2 Spike RBD (Arg319-Phe541)(Tyr453Phe) Accession # YP_009724390.1	IEGRMD	Human IgG₁ (Pro100-Lys330)
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

Protein identity is confirmed by mass spectrometry.

Predicted Molecular Mass

52 kDa

SDS-PAGE

57-64 kDa, under reducing conditions.

Activity

Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag (Catalog # 933-ZN).

Recombinant SARS-CoV-2 Y453F Spike RBD Fc Chimera Protein CF Scientific Data Examples

	Recombinant SARS-CoV-2 Y453F Spike RBD Fc Chimera Protein Binding Activity. Recombinant SARS-CoV-2 Y453F Spike RBD Fc Chimera (Catalog # 10792-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.
	Recombinant SARS-CoV-2 Y454F Spike RBD Fc Chimera Protein SDS-PAGE. 2 μg/lane of Recombinant SARS-CoV-2 Y454F Spike RBD Fc Chimera (Catalog # 10792-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 57-64 kDa and 114-128 kDa, respectively.
	Binding of ACE-2 to SARS-CoV-2 Spike RBD protein Y453F mutant by surface plasmon resonance (SPR). Recombinant SARS-CoV-2 Spike RBD protein Y453F mutant Fc-tag (Catalog # 10792-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 94.3 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 K_D=1.33 nM. (Biacore T200).

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

10792-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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 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 (ACE-2), 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% amino acid (aa) identity with the RBD of the SARS-CoV-1, but only 22% aa identity with the RBD of MERS. SARS-CoV-2 variants carrying the aa substitution Y453F in the RBD was reported to escape neutralization of antibodies (8).

Item	Value
References	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. Li, W. et al. (2003) Nature 426:450. Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197. Hoffman, M. et al. (2021) Cell Reports. 35:109017.
Long Name	Spike Receptor Binding Domain
Entrez Gene IDs	3200426 (HCoV-HKU1); 14254594 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)
Alternate Names	Spike RBD

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