General description
Spike glycoprotein (UniProt: K9N5Q8; also known as S glycoprotein, E2, Peplomer protein) is encoded by the S (also known as 3) gene (Gene ID: 14254594) in Middle East respiratory syndrome-related coronavirus (MERS-CoV). MERS-CoV is a positive-strand RNA virus of the betacoronavirus genus within the Coronaviridae family that causes severe respiratory syndrome in human. The mature MERS-CoV contains 4 structural proteins: Envelope (E), Membrane (M), Nucleocapsid (N), and the Spike protein (S). Spike protein is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-17), which is subsequently cleaved off to generate the mature form that can be cleaved into three subunits known as S1 (aa 18-751), S2 (aa 752-1353), and S2 (aa 888-163) by host cell furin or other cellular proteases. Spike protein has an extracellular domain (aa 18-1296), a transmembrane domain (aa 1297-1317), and a cytoplasmic domain (aa 1318-1353). S1 subunit attaches the virion to the cell membrane by interacting with host receptor, initiating the infection. It interacts with host DPP4 to mediate viral entry. The S2 subunit mediates fusion of the virion and cellular membranes by acting as a class I viral fusion protein. Clone m336 binds to the receptor binding domain of spike protein that is localized in amino acids 360-601. It is shown to neutralized both pseudotyped and live MERS-CoV with high potency likely by competing with DPP4 for binding to the S glycoprotein. It does not exhibit reactivity with SARS-CoV2 spike protein. (Ref.: Ying, T., et al. (2014). J. Virol. 88(14); 7796-7805).
Specificity
Clone m336 is a recombinant human monoclonal antibody that detects spike protein in Middle East respiratory syndrome-related coronavirus (MERS-CoV). It targets an epitope within the receptor binding domain.
Immunogen
Generated with a large phage display library constructed by using cDNA of peripheral blood mononuclear cells (PBMC) from healthy volunteers and then panned using the RBD (360-601) of MERS-CoV.
Application
Quality Control Testing
Evaluated by ELISA with recombinant receptor binding domain (RBD) from spike protein of MERS-CoV.
ELISA Analysis (ELISA): Various dilutions of this antibody detected recombinant receptor binding domain (RBD) from spike protein of MERS-CoV.
Tested Applications
Neutralizing: A representative lot neutralized MERS-CoV RBD in Neutralizing applications (Ying, T. et al. (2014). J Virol.;88(14):7796-805; Folegatti, P.M., et al. (2020). Lancet Infect Dis.;20(7):816-826).
ELISA Analysis: A representative lot detected MERS-CoV RBD in ELISA applications (Ying, T. et al. (2014). J Virol.;88(14):7796-805; Folegatti, P.M., et al. (2020). Lancet Infect Dis.;20(7):816-826).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Anti-MERS-CoV RBD, clone m336, Cat. No. MABF3060, is a human recombinant monoclonal antibody that detects of Spike glycoprotein of Middle East respiratory syndrome-related coronavirus and is tested for use in ELISA and Neutralizing.
Physical form
Purified human recombinant monoclonal antibody IgG1 in PBS without preservatives.
Storage and Stability
Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.