Human Anti-EBOV GP Recombinant Antibody (clone mAb114)
CAT#: FAMAB-0525WJ
This product is a human monoclonal antibody reacted with EBOV GP. This antibody can be used in a variety of applications, such as ELISA, WB, Neut, IP.
Published Data
Neut
Figure 1 Neutralization of vaccination-induced macaque monoclonal antibodies (mAbs).
Bar chart representing the values of half-maximal inhibitory concentration ([IC50] μg/mL) calculated after in vitro neutralization assay using Zaire ebolavirus (EBOV) glycoprotein (GP)-pseudotyped-virus performed with the 14 mAbs that were cloned. In gray are negative controls VRC01 and mAb114 used as a reference. Seven of 14 mAbs showed ability to neutralize in vitro.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
Neut
Figure 2 Neutralization of vaccination-induced macaque monoclonal antibodies (mAbs).
Dot plots show the-log half-maximal effective concentration ([EC50] μg/mL) representing binding of the 7 neutralizers identified in A to GP (filled circles) and GPΔMUC (empty circles). All of the mAbs tested bound GPΔMUC with greater potency compared with GP (P = .0097).
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
IP
Figure 3 Immunoprecipitation of the 7 neutralizers performed with GPTHL.
Four of seven mAbs were able to precipitate GPTHL. As control, input GPTHL is shown on a Coomassie-stained gel.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
WB
Figure 4 Western blot on the 4 mAbs identified in C performed with GPΔMUC and GPTHL.
For mAb114, none of the mAbs tested were able to bind GPTHL by Western blot. As a positive control for GPTHL, binding of polyclonal serum is shown.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
Inhib
Figure 5 Bar chart representing the percentage of binding inhibition of biotinylated mAb114 to GPΔMUC in the presence of each of the 4 putative mAb114-like antibodies.
All mAbs showed a high degree of competition with mAb114 for binding to GPΔMUC (calculated at the concentration of 8 μg/mL). The gray bar shows the maximal degree of self-competition of mAb114.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
Figure 6 Bar chart on the binding kinetics of the GP1 core antibodies compared with mAb114.
log (KD) values were calculated by biolayer interferometry performed with Fab binding to GPΔMUC at neutral and low pH (black and dark gray bars, respectively) as well as to GPTHL at low pH only (light gray bars). All monoclonal antibodies (mAbs) showed similar kinetics compared with mAb114 with ma-D08 showing slightly lower values. Fab fragments derived from papain digestion of mAbs were used for measuring kinetics. This experiment was performed twice. Values are shown from 1 representative experiment.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
Figure 7 Bar chart on competition of the GP1 core antibodies with NPC1-dC compared with mAb114 performed by biolayer interferometry using whole IgG binding to GPTHL.
All mAbs showed similar competition with NPC1-dC for binding to GPTHL. Human immunodeficiency virus GP120 (indicated as "Control") and 2 additional mAbs (KZ52 and mAb100) that are known to bind the base of GPTHL far from the receptor binding domain (RBD) were used as negative controls. NPC1-dC was used to assess the maximal degree of self-competition. This experiment was also performed twice, and values are shown from 1 representative experiment.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
Figure 8 Electron microscopy reveals that all of the RBD blockers approach glycoprotein (GP) vertically but with a slightly more open angle compared with mAb114 (Fab indicated with green stars).
The scale bars are 10 nm.
Cagigi, A., Misasi, J., Ploquin, A., Stanley, D. A., Ambrozak, D., Tsybovsky, Y., ... & Sullivan, N. J. (2018). Vaccine generation of protective Ebola antibodies and identification of conserved B-cell signatures. The Journal of Infectious Diseases, 218(suppl_5), S528-S536.
❮
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Specifications
- Host Species
- Human
- Type
- Human IgG
- Specificity
- EBOV GP
- Species Reactivity
- EBOV
- Clone
- mAb114
- Applications
- ELISA, WB, Neut, IP
- Related Disease
- EBOV infection
Product Property
- Purity
- >95% as determined by SDS-PAGE
- Concentration
- Please refer to the vial label for the specific concentration.
- Buffer
- PBS
- Preservative
- No preservatives
- Storage
- Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.
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Consistent Storage
The antibody is stable and effective for a considerable amount of time after it is reconstituted. This has proven to be highly advantageous for our extended projects.
Friendly to Users
Everything about the experience has gone smoothly, from ordering to actually using in the lab. The outcomes speak for themselves, and handling the antibodies is simple.
Comprehensive Records
This antibody comes with thorough and informative documentation. Our approach is now lot simpler thanks to usage guidelines and instructions for dilutions.
Q&As
-
In what way should I employ the antibody to maintain its potency?
A: Avoid repeated cycles of freezing and thawing, aliquot as needed, and store as directed.
-
Does optimizing the protocol become necessary?
A: While the antibody works well in standard conditions, there may be a few small tweaks that need to be made for your specific experimental setup.
-
Exist any shipping safety precautions?
A: The antibody is shipped on dry ice for stability. As soon as you receive it, make sure to preserve it according to the instructions.
View the frequently asked questions answered by Creative Biolabs Support.
Cite This Product
To accurately reference this product in your publication, please use the following citation information:
(Creative Biolabs Cat# FAMAB-0525WJ, RRID: AB_3111330)
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Downloadable Resources
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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.
Datasheet
MSDS
COA
Certificate of Analysis LookupTo download a Certificate of Analysis, please enter a lot number in the search box below. Note: Certificate of Analysis not available for kit components.
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