Recombinant Human Anti-SARS S Antibody (80R) (CAT#: PABZ-139)

Recombinant Human Antibody (80R) is capable of binding to SARS S, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-SARS S mAb and CH1-3 region of human IgG and a light chain (LC) encoding VL from anti-SARS S proteins mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition. We show that the 80R-binding epitope on the S1 RBD overlaps very closely with the ACE2 binding site, providing a rationale for the strong binding and broad neutralizing ability of the antibody.

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Figure 1 Microneutralization assay of anti-S1 antibodies on SARS-CoV.

Figure 1 Microneutralization assay of anti-S1 antibodies on SARS-CoV.

(A) Microneutralization assay of anti-S1 scFvs. The positive control was convalescent serum from a SARS patient; the negative control was non-SARS human serum. The names of the scFvs are labeled at the top, and antibody titers are indicated on the left. Neutralizing scFv 80R and only one nonneutralizing scFv 27D (illustrative of the other nonneutralizing scFvs) are shown. Undiluted SARS-CoV ( ~37 plaque-forming units) was loaded per well. (B) Comparison of the neutralization activity of 80R scFv and full-length 80R IgG1. The positive and negative control serum samples and the amount of virus used were the same as in A. The titer and concentration of antibodies are labeled on the left.

Sui, J., Li, W., Murakami, A., Tamin, A., Matthews, L. J., Wong, S. K., ... & Anderson, L. J. (2004). Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association. Proceedings of the National Academy of Sciences, 101(8), 2536-2541.

Figure 2 Kinetic rates and binding affinity of 80R antibody and ACE2 to S1-lg

Figure 2 Kinetic rates and binding affinity of 80R antibody and ACE2 to S1-lg

Sui, J., Li, W., Murakami, A., Tamin, A., Matthews, L. J., Wong, S. K., ... & Anderson, L. J. (2004). Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association. Proceedings of the National Academy of Sciences, 101(8), 2536-2541.

Figure 3 Inhibition of syncytia formation by anti-S1 antibodies.

Figure 3 Inhibition of syncytia formation by anti-S1 antibodies.

Shown is syncytia formation assay with anti-S1 antibodies. 293T cells expressing SARS-CoV S protein were preincubated with the indicated concentrations of anti-S1 scFvs or 80R IgG1 and then mixed with 293T cells expressing ACE2. After culturing for 36 h in the presence of antibodies, dose-dependent inhibition of syncytia formation by 80R scFv, 80R IgG1 was observed and photographed. Representative results are shown.

Sui, J., Li, W., Murakami, A., Tamin, A., Matthews, L. J., Wong, S. K., ... & Anderson, L. J. (2004). Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association. Proceedings of the National Academy of Sciences, 101(8), 2536-2541.

Figure 4 80R scFv inhibiting the binding of S1 to ACE2 receptor.

Figure 4 80R scFv inhibiting the binding of S1 to ACE2 receptor.

(A) Flow cytometry histograms; shown is staining Vero E6 with S1-Ig and flow cytometry analysis. Dotted line, control staining with S1 (327)-Ig; thin line, cells were stained with S1-Ig; bold line, staining with premix of 0.3 μg of S1-Ig and 0.3 μg of 80R scFv (Left) or 27D scFv (Right). (B) scFv competition of S1-Ig binding to ACE2 in immunoprecipitation. Radiolabeled ACE2 was immunoprecipitated by S1-Ig that was preincubated with the indicated amounts of either 27D scFv or 80R scFv. Anti-ACE2 precipitates were used as a positive control. Immunoprecipitates were run on a reducing SDS-PAGE gel and visualized by autoradiography.

Sui, J., Li, W., Murakami, A., Tamin, A., Matthews, L. J., Wong, S. K., ... & Anderson, L. J. (2004). Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association. Proceedings of the National Academy of Sciences, 101(8), 2536-2541.


Specifications

  • Immunogen
  • SARS Spike glycoprotein
  • Host Species
  • Human
  • Derivation
  • Human
  • Type
  • IgG
  • Specificity
  • Tested positive against native SARS S
  • Species Reactivity
  • SARS
  • Clone
  • 80R
  • Applications
  • Neut Assay-Dependent
    Inhib Assay-Dependent

Product Property

  • Purity
  • >95% by SDS-PAGE and HPLC analysis
  • Storage
  • Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing of samples.

Applications

  • Application Notes
  • The antibody 80R has been reported in applications of Neut, Inhib. It's recommended that the optimal antibody concentration, dilution, incubition time etc. are best to be carefully titrated in specific assays.

Target

  • Alternative Names
  • Spike glycoprotein; S protein

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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For Research Use Only. Not For Clinical Use.

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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