Name: Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK)
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Data and images
Specifications

Product data and images for Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK)

Figures

Figure 1

**Cell-based RBD binding assay**
. Schematic overview illustrating how our biotinylated RBD can be applied to analyze binding to ACE2-expressing cells.
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 monomer is biologically functional and shows a titratable, high-affinity binding to cell surface ACE2.

Figure 1

Cell-based RBD binding assay

. Schematic overview illustrating how our biotinylated RBD can be applied to analyze binding to ACE2-expressing cells.

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 monomer is biologically functional and shows a titratable, high-affinity binding to cell surface ACE2.

Figure 2

**Results of the cell-based binding assay using biotinylated Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK).**
ACE2-expressing cells (Vero E6 cells) were incubated with biotinylated recombinant SARS-CoV-2 RBD of the wild-type, Wuhan variant (Recombinant SARS-CoV-2 RBD (HEK)-Biotin) and the SARS-CoV-2 variant B.1.1.529/BA.1, Omicron (Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1(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 RBD is biologically functional and shows a titratable, high-affinity binding to cell surface ACE2.

Figure 2

Results of the cell-based binding assay using biotinylated Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK).

ACE2-expressing cells (Vero E6 cells) were incubated with biotinylated recombinant SARS-CoV-2 RBD of the wild-type, Wuhan variant (Recombinant SARS-CoV-2 RBD (HEK)-Biotin) and the SARS-CoV-2 variant B.1.1.529/BA.1, Omicron (Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1(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 RBD is biologically functional and shows a titratable, high-affinity binding to cell surface ACE2.

Figure 3

**SDS-PAGE of biotinylated Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK)**
under reduced (R) and non-reduced (NR) conditions.

Figure 3

SDS-PAGE of biotinylated Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK)

under reduced (R) and non-reduced (NR) conditions.

Figure 4

**Size-exclusion chromatography (SEC) Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK).**
The non-biotinylated protein was analyzed and the peak corresponds to the monomeric protein.

Figure 4

Size-exclusion chromatography (SEC) Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK).

The non-biotinylated protein was analyzed and the peak corresponds to the monomeric protein.

Specifications for Recombinant SARS-CoV-2 RBD B.1.1.529/BA.1 (HEK)

Overview

The receptor binding domain (RBD) locates C-terminally within the S1 subunits of the spike (S) protein. The S protein forms a homotrimeric structure on the surface of the SARS-CoV-2 virus and binds to the angiotensin-converting enzyme 2 (ACE2) receptor of target cells. Recent studies have shown that the predominant activated form displays one RBD rotated upwards into an ACE2-accessible state and it is known that binding of RBD to ACE2 is a major contributor for interaction of the S protein with its target receptor.

The SARS-CoV-2 mutant strain B.1.1.529, also known as Omicron variant, was first identified during the Covid-19 pandemic in Botswana/ South Africa in late November 2021. Due to increased transmissibility and substantial immune evasion it spread rapidly around the globe.

The BA.1 lineage is defined by 16 amino acid substitutions in the RBD (G339D, S371L, S373P, S375F, K417N (already known from B.1.351-Beta), N440K, G446S, S477N, T478K (already known from B.1.617.2-Delta), E484A, Q493R, G496S, Q498R, N501Y (already known from B.1.1.7-Alpha, B.1.351-Beta and P.1-Gamma), T547K, Y505H).

Preliminary evidence suggests an increased risk of reinfection with this variant, as compared to other VOCs.

The SARS-CoV-2 RBD B.1.1.529 (HEK) protein covers amino acids R319 to S591 of the spike protein and contains a C-terminal His-tag and a N-terminal AviTag. Recombinant SARS-CoV-2 RBD B.1.1.529 (HEK)-Biotin is specifically biotinylated at a single site, preserving full functionality of the RBD.

Preliminary evidence suggests an increased risk of reinfection with this variant, as compared to other VOCs.

Alternative names

SARS-CoV-2 Omicron variant

Data and images