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SARS-CoV-2 Spike S1 Proteins

Spike-S1-His Unit size Cat. code Docs Qty Price
SARS-CoV-2 Spike S1-His fusion protein
50 µg
his-sars2-s1
+-
$210.00
Spike-S1-Fc Unit size Cat. code Docs Qty Price
SARS-CoV-2 Spike S1-Fc fusion protein
50 µg
fc-sars2-s1
+-
$210.00

SARS-CoV-2 Spike S1 subunit with C-term His- or Fc-tag

PROTEIN DESCRIPTION

Potential applications of soluble tagged Spike S1 proteins
Potential applications of soluble tagged Spike S1 proteins

The SARS-CoV-2 (2019-nCoV) Spike S1 subunit plays a crucial role in the viral entry into the target cell. The S1 subunit features an N-terminal domain (S1-NTD)  and a C-terminal domain (S1-CTD). The S1-NTD is thought to mediate sugar-binding and the S1-CTD allows the virus to bind to ACE2 through the receptor-binding domain (RBD) [1-3]. In its resting conformation, S1 exerts a physical constraint on the Spike fusion subunit [3]. Ongoing research is attempting to understand the exact mechanisms that drive conformation changes in S1 allowing subsequent membrane fusion events. S1 is a candidate for subunit vaccines against SARS-CoVs [4, 5].

Spike-S1-His and Spike-S1-Fc were generated by fusing the SARS-CoV-2 Spike S1 [V16-R685] to a C-terminal poly-histidine sequence and human IgG1 Fc region, respectively. Of note, the SARS-CoV-2 viral sequence used is from the Wuhan-Hu-1 (D614) isolate. 

Spike-S1-His and Spike-S1-Fc have been produced in HEK293 cells and CHO cells, respectively, and have been purified by affinity chromatography (See Specifications for more information).

 

APPLICATIONS

  • Vaccination studies: using combinations of Spike protein antigens and adjuvants
  • Antibody screening: finding anti-Spike antibodies that can neutralize the SARS-CoV-2 infection
  • Inhibitor screening: finding small molecules or antibodies able to block the SARS-CoV-2 RBD interaction with the ACE2 receptor
  • ACE2 cellular expression screening: in primary isolated cells or transfected cells

QUALITY CONTROL

 

Learn more on SARS-CoV-2Learn more about SARS-CoV-2 infection cycle, immune responses, and potential therapeutics.

 

References

1. Li F., 2016. Structure, function, and evolution of coronavirus spike proteins. Annu. Rev. Virol. 3:237-261.
2. Li F. et al., 2005. Structure of SARS coronavirus spike receptor-binding domain complexed with receptor. Science. 309:1864-1868.
3. Walls A.C. et al., 2020. Structure, function, and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell. 181(2):281-292.e6.
4. Wang N. et al., 2020. Subunit vaccines against emerging pathogenic human coronaviruses. Front. Microbiol. 11:298. DOI: 10.3389/fmicb.2020.00298.
5. Padron-Regalado E., 2020. Vaccines for SARS-CoV-2: Lessons from other coronavirus strains. Infect. Dis. Ther. DOI: 10.1007/s40121-020-00300-x.

Figures

Spike-S1-His purity analysis by SDS PAGE
Spike-S1-His purity analysis by SDS PAGE

SDS PAGE analysis of the SARS-CoV-2 Spike-S1-His protein.

2 μg of the fusion protein was loaded onto a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per manufacturer’s instructions.

Recognition of Spike-S1-His by anAnti-SARS-CoV-Spike (CR3022) human IgM
Recognition of Spike-S1-His by anAnti-SARS-CoV-Spike (CR3022) human IgM

ELISA detection of Spike-S1-His fusion protein with an Anti-SARS-CoV spike human IgM (CR3022).

Anti-SARS-CoV-Spike hIgM antibody (5 μg/ml) was coated onto ELISA plates overnight. Following this, a 3-fold serial dilution of Spike-S1-His (red curve) or control protein (ACP5-His; grey curve) were added and incubated for 1 hour. Binding was detected using a HRP-labelled anti-His antibody (1/1000 dilution) and the HRP substrate OPD (o-phenylenediamine dihydrochloride). Absorbance was read at 490 nm.

Spike-S1-Fc purity analysis by SDS PAGE
Spike-S1-Fc purity analysis by SDS PAGE

SDS PAGE analysis of the SARS-CoV-2 Spike-S1-Fc protein.

2 μg of the fusion protein was loaded onto a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per manufacturer’s instructions

Recognition of Spike-S1-Fc by anAnti-SARS-CoV-Spike (CR3022) human IgM
Recognition of Spike-S1-Fc by anAnti-SARS-CoV-Spike (CR3022) human IgM

ELISA detection of Spike-S1-Fc fusion protein with an Anti-SARS-CoV spike human IgM (CR3022).

Anti-SARS-CoV-Spike hIgM antibody (5 μg/ml) was coated onto ELISA plates overnight. Following this, a 3-fold serial dilution of Spike-S1-Fc (red curve) or control protein (CTLA4-Fc; grey curve) were added and incubated for 1 hour. Binding was detected using a HRP-labelled anti-His antibody (1/1000 dilution) and the HRP substrate OPD (o-phenylenediamine dihydrochloride). Absorbance was read at 490 nm.

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Specifications

Spike-S1-His

  • Protein construction: S1 [V16-R685] from the Spike glycoprotein with a C-terminal poly-histidine tag 
  • Accession sequence: YP_009724390
  • Species: SARS-CoV-2 (2019-nCoV); Wuhan-Hu-1 (D614) isolate
  • Tag: C-terminal poly-histidine (6 x His)
  • Total protein size: 681 a.a. (secreted form)
  • Molecular weight: ~ 123 kDa (SDS-PAGE gel)
  • Purification: Ni2+ affinity chromatography
  • Purity: >95% (SDS-PAGE)
  • Quality control:
    - The protein has been validated by ELISA upon incubation with a coated Anti-SARS-CoV-Spike-RBD hIgM mAb (clone CR3022)
    - The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cellular assays.

Spike-S1-Fc

  • Protein construction: S1 [V16-R685] from the Spike glycoprotein with a C-terminal human IgG1 Fc tag 
  • Accession sequence: YP_009724390
  • Species: SARS-CoV-2 (2019-nCoV); Wuhan-Hu-1 (D614) isolate
  • Tag: C-terminal human IgG1 Fc
  • Total protein size: 920 a.a. (secreted form)
  • Molecular weight: ~124 kDa (SDS-PAGE)
  • Purification: Protein G affinity chromatography
  • Purity: >95% (SDS-PAGE)
  • Quality control:
    - The protein has been validated by ELISA upon incubation with a coated Anti-SARS-CoV-Spike-RBD hIgM mAb (clone CR3022) 
    - The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cellular assays.
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Contents

Spike-S1-His and Spike-S1-Fc contents:

  • 50 μg of lyophilized protein
  • 1.5 ml of endotoxin-free water

room temperature The product is shipped at room temperature.

store Lyophilized protein should be stored at -20 ̊C.

stability Resuspended protein is stable up to 1 month when stored at 4°C, and 1 year when stored at -20°C

Avoid repeated freeze-thaw cycles.

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Details

Learn more about the SARS-CoV-2 Spike protein here.

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