Alternative names:
SARS-CoV-2 Beta variant

Data and images for Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK)

Figures

Figure 1

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Cell-based spike protein binding assay.
Schematic overview illustrating how our biotinylated spike proteins can be applied to analyze binding to ACE2-expressing cells.

Figure 1

Cell-based spike protein binding assay.
Schematic overview illustrating how our biotinylated spike proteins can be applied to analyze binding to ACE2-expressing cells.

Figure 2

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Results of the cell-based binding assay using biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK).
ACE2-expressing cells (Vero E6 cells) were incubated with biotinylated SARS-CoV-2 spike monomer of the wild-type, Wuhan variant (Recombinant SARS-CoV-2 Spike-Prot (HEK)-Biotin), B.1.1.7, Alpha (Recombinant SARS-CoV-2 Spike-Prot B.1.1.7 (HEK)-Biotin), and B.1.351, Beta (Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK)-Biotin).
Different protein concentrations were used, and ACE2-bound spike protein was subsequently detected by fluorescent staining with streptavidin-PE. Quantification of stained cells at different protein concentrations illustrates that Miltenyi Biotec's recombinant spike monomers are biologically functional and show a titratable, high-affinity binding to cell surface ACE2. B.1.351 (HEK) displayed the second highest binding affinity to ACE2.

Figure 2

Results of the cell-based binding assay using biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK).
ACE2-expressing cells (Vero E6 cells) were incubated with biotinylated SARS-CoV-2 spike monomer of the wild-type, Wuhan variant (Recombinant SARS-CoV-2 Spike-Prot (HEK)-Biotin), B.1.1.7, Alpha (Recombinant SARS-CoV-2 Spike-Prot B.1.1.7 (HEK)-Biotin), and B.1.351, Beta (Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK)-Biotin).
Different protein concentrations were used, and ACE2-bound spike protein was subsequently detected by fluorescent staining with streptavidin-PE. Quantification of stained cells at different protein concentrations illustrates that Miltenyi Biotec's recombinant spike monomers are biologically functional and show a titratable, high-affinity binding to cell surface ACE2. B.1.351 (HEK) displayed the second highest binding affinity to ACE2.

Figure 3

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SDS-PAGE of biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK)
under reduced (R) and non-reduced (NR) conditions

Figure 3

SDS-PAGE of biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK)
under reduced (R) and non-reduced (NR) conditions

Specifications for Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK)

Overview

The SARS-CoV-2 spike glycoprotein (S) mediates the entry of the virus into target cells via its interaction with the angiotensinconverting enzyme 2 (ACE2) receptor, thereby initiating the infection. It forms a homotrimeric structure on the surface of the SARS-CoV-2 virus and represents the major target for diagnostic and therapeutic agents. The ectodomain of the S glycoprotein comprises the N-terminal S1 subunit, including the receptor binding domain (RBD) and the C-terminal S2 subunit. The ectodomain spans from aa12 to aa1213 followed by a transmembrane helix beginning at aa1214–aa1234. The C-terminus of the native protein is formed by a cytoplasmic domain spanning aa1235–aa1273.
The SARS-CoV-2 mutant strain B.1.351, also known as 501Y.V2 or variant Beta, was first identified in of South Africa in December 2020. Due to increased transmissibility it spread rapidly around the globe. The B.1.351 lineage (Beta variant) differs from the wildtype sequence by a total of 21 amino acid mutations of which three key amino acid substitutions K417N, E484K, and N501Y occur in the RBD. The N501Y mutation also emerged independently in SARS-CoV-2 variants of the B.1.1.7 lineage (Alpha variant) and P.1 lineage (Gamma variant). The B.1.351 lineage (Beta variant) is reported to have a higher infectivity due to the increased affinity to the ACE2 receptor and shows reduced susceptibility to neutralizing antibodies.
The Recombinant SARS-CoV-2 Spike-Prot B.1.351 (HEK) protein covers the ectodomain of the viral surface protein including amino acids V16 to K1211. It was engineered to contain the stabilizing proline substitutions at position K986P and V987P. The native sequence of the Furin cleavage site (RRAR at residues 682-685) was substituted by GSAS to further increase recombinant protein stability.
In addition, the SARS-CoV-2 Spike-Prot B.1.351 (HEK) protein contains the amino acid substitutions L18F, D80A, D215G, R246I, K417N, E484K, N501Y, D614G, and A701V. The protein is extended at its C-terminus with a His-tag and an AviTag™. Recombinant SARSCoV- 2 Spike-Prot B.1.351 (HEK)-Biotin is specifically biotinylated at a single site, preserving full functionality of the protein.

Alternative names

SARS-CoV-2 Beta variant

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