SARS-CoV-2 (2019-nCoV) Spike Protein (S2 ECD, His tag) (100 μg)

Purity: >90 % as determined by SDS-PAGE.
Tag: His-tag at the C-terminus.
Endotoxin: <1.0 EU per μg protein as determined by the LAL method.
Activity: Testing in progress.
Protein Construction: A DNA sequence encoding the NCP-CoV(2019-nCoV) Spike Protein (S2 ECD) was expressed with a polyhistidine tag at the C-terminus.
Expressed Host: Baculovirus-Insect Cells.
Species: 2019-nCoV
Predicted N Terminal: Ser
Predicted Molecular Mass: SARS-CoV-2 (2019-nCoV) Spike Protein (S2 ECD, His tag) consists of 539 amino acids and predicts a molecular mass of 59.37 kDa.
2019-nCov amino acids: Ser686 – Pro1213 (Gene Accession Number: YP_009724390.1)
Formulation: Lyophilized from sterile 20 mM PB, 300 mM NaCl, 10 % glycerol, pH 7.0.Normally 5 % – 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
Shipping: In general, recombinant proteins are provided as lyophilized powder which are shipped at ambient temperature.
Bulk packages of recombinant proteins are provided as frozen liquid. They are shipped out with blue ice unless customers require otherwise.
Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store under sterile conditions. It is recommended that the protein be aliquoted for optimal storage. Repeated freeze/thaw cycles should be avoided.
Regulatory/Restrictions: For Research Use Only.

SARS-CoV-2 (2019-nCoV) Spike Protein (S2 ECD, His tag) Datasheet

In late December 2019, a number of patients with viral pneumonia (now called 2019-nCoV Acute Respiratory Disease) were found to be epidemiologically associated with the Huanan seafood market in Wuhan, in the Hubei province of China. A novel, human-infecting coronavirus, provisionally named 2019 Novel Coronavirus (2019-nCoV), and since named SARS-CoV-2, was identified by genomic sequencing (Lu et al., 2020). SARS-CoV-2 is closely related (88% identity) to two bat-derived severe acute respiratory syndrome (SARS)-like coronaviruses, collected in 2018 in Zhoushan, eastern China, but were more distant from SARS-CoV (~79% identity) and MERS-CoV (~50% identity). However, although bats might be the original host of this virus, an animal sold at the seafood market in Wuhan may have acted as an intermediate host (Lu et al., 2020). Relative synonymous codon usage (RSCU) analysis suggests that SARS-CoV-2 is a recombinant between the bat coronavirus and an origin-unknown coronavirus, and it has been proposed that a pangolin could have acted as the reservoir. The recombination event occurred within the viral Spike glycoprotein (S) (Ji et al., 2020).

Homology modelling shows that SARS-CoV-2 has a similar receptor-binding domain structure to that of SARS-CoV, despite amino acid variation at some key residues. Therefore, SARS-CoV-2 may be able to bind to the angiotensin-converting enzyme 2 (ACE2) receptor in humans (Lu et al., 2020).

The virus spreads primarily through contact with an infected person, through respiratory droplets generated when a person coughs or sneezes, or through droplets of saliva or discharge from the nose. The incubation period is believed to range from 2-11 days. Infection with SARS-CoV-2 can cause mild symptoms including a runny nose, sore throat, cough, and fever. However, it can be more severe for some people and can lead to pneumonia or breathing difficulties. The elderly, and people with pre-existing medical conditions -such as, diabetes and heart disease- appear to be more vulnerable to becoming severely ill with the virus. There are currently more than 40,000 confirmed cases from 24 countries, although the vast majority are still within China, with more than 900 deaths to date (WHO, 2020).

The coronavirus Spike (S) glycoprotein is a class I viral fusion protein on the outer envelope of the virion that plays a critical role in viral infection by recognizing host cell receptors and mediating fusion of the viral and cellular membranes (Li, 2016). The S glycoprotein is synthesized as a precursor protein consisting of ~1,300 amino acids that is then cleaved into an amino (N)-terminal S1 subunit (~700 amino acids) and a carboxyl (C)-terminal S2 subunit (~600 amino acids). Three S1/S2 heterodimers assemble to form a trimer spike protruding from the viral envelope. The S1 subunit contains a receptor-binding domain (RBD), while the S2 subunit contains a hydrophobic fusion peptide and two heptad repeat regions. Triggered by receptor binding, proteolytic processing and/or acidic pH in the cellular compartments, the class I viral fusion protein undergoes a transition from a metastable prefusion state to a stable postfusion state during infection, in which the receptor-binding subunit is cleaved, and the fusion subunit undergoes large-scale conformational rearrangements to expose the hydrophobic fusion peptide, induce the formation of a six-helix bundle, and bring the viral and cellular membranes close for fusion (Belouzard et al., 2012). The trimeric SARS coronavirus (SARS-CoV) S glycoprotein consisting of three S1-S2 heterodimers binds the cellular receptor angiotensin-converting enzyme 2 (ACE2) and mediates fusion of the viral and cellular membranes through a pre- to postfusion conformation transition (Song et al., 2018).