Human Anti-ENV Recombinant Antibody (clone 2F5) (CAT#: PABZ-141)

Recombinant Human Antibody (2F5) is capable of binding to SIV gp41, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-SIV gp41 mAb and CH1-3 region of human IgG and a light chain (LC) encoding VL from anti-SIV gp41 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. The binding site for the neutralizing antibody 2F5 occurs at the end of the C-terminal helix in a region which is solvent exposed and proximal to the fusion peptide.


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Figure 1 SHIV neutralization by 2F5, 4E10, and b12 in a rhesus PBMC assay and a pseudovirus luciferase reporter gene assay

Figure 1 SHIV neutralization by 2F5, 4E10, and b12 in a rhesus PBMC assay and a pseudovirus luciferase reporter gene assay

Hessell, A. J., Rakasz, E. G., Tehrani, D. M., Huber, M., Weisgrau, K. L., Landucci, G., ... & Burton, D. R. (2010). Broadly neutralizing monoclonal antibodies 2F5 and 4E10 directed against the human immunodeficiency virus type 1 gp41 membrane-proximal external region protect against mucosal challenge by simian-human immunodeficiency virus SHIVBa-L. Journal of virology, 84(3), 1302-1313.

Figure 2 Protection by 2F5 and 4E10 against mucosal SHIV<sub>Ba-L</sub> challenge.

Figure 2 Protection by 2F5 and 4E10 against mucosal SHIVBa-L challenge.

A total of 16 male Indian rhesus macaques were divided into two antibody treatment groups of six for administration of 2F5 and 4E10. The control group was comprised of two isotype control-treated animals and two control animals (96068 and 2025) that were challenged prior to the beginning of the protection study to confirm viral infectivity but not treated with antibody. All animals were intrarectally challenged with 2,000 TCID₅₀ SHIVBa-L. Animals in the antibody treatment groups received an intravenous dose of 50 mg/kg of either 2F5, 4E10, or isotype control antibody 1 day prior to viral challenge (day - 1) and again 1 day following viral challenge (day + 1). All control animals (4/4) experienced peak viremia on the order of 107 between days 14 and 21. A single animal (2F5-treated 93060), experienced very low-level transient viremia, but at the end of 6 weeks there was no detectable virus in any of the animals. The presence of virus was measured in an assay with a minimum detection limit of 150 SIV RNA copies per ml (2.1 log₁₀ vRNA copies/ml) with a 95% confidence level. A second assay with a minimum detection limit of 10 SIV RNA copies per ml was used to retest available samples of sufficient volume (Table 2).

Hessell, A. J., Rakasz, E. G., Tehrani, D. M., Huber, M., Weisgrau, K. L., Landucci, G., ... & Burton, D. R. (2010). Broadly neutralizing monoclonal antibodies 2F5 and 4E10 directed against the human immunodeficiency virus type 1 gp41 membrane-proximal external region protect against mucosal challenge by simian-human immunodeficiency virus SHIVBa-L. Journal of virology, 84(3), 1302-1313.

Figure 3 Antibody-dependent cell-mediated viral inhibition (ADCVI) of 2F5 and 4E10 in comparison to b12.

Figure 3 Antibody-dependent cell-mediated viral inhibition (ADCVI) of 2F5 and 4E10 in comparison to b12.

Target cells (CEM.NKRCCR5) were infected with SHIVBa-L, incubated for 48 h, and then washed to remove cell-free virus. Serially diluted antibody was added at 50, 2, and 0.4 μg/ml to effector cells (rhesus PBMCs) and incubated for 30 min. Effectors and diluted antibody were combined with targets at an effector-to-target ratio of 10:1 and incubated for 7 days at 37°C in 5% CO2. Supernatant was collected and assayed for p27 by ELISA to measure viral inhibition.

Hessell, A. J., Rakasz, E. G., Tehrani, D. M., Huber, M., Weisgrau, K. L., Landucci, G., ... & Burton, D. R. (2010). Broadly neutralizing monoclonal antibodies 2F5 and 4E10 directed against the human immunodeficiency virus type 1 gp41 membrane-proximal external region protect against mucosal challenge by simian-human immunodeficiency virus SHIVBa-L. Journal of virology, 84(3), 1302-1313.


Specifications

  • Immunogen
  • SIV envelope protein
  • Host Species
  • Human
  • Type
  • Human IgG
  • Specificity
  • SIV E protein
  • Species Reactivity
  • SIV, HIV1
  • Clone
  • 2F5
  • Applications
  • Neut

Product Property

  • Purity
  • >95% as determined by SDS-PAGE and HPLC analysis
  • Concentration
  • Please refer to the vial label for the specific concentration.
  • 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.

Applications

  • Application Notes
  • The antibody 2F5 has been reported in applications of Neutralization, Inhibition. It's recommended that the optimal antibody concentration, dilution, incubition time etc. are best to be carefully titrated in specific assays.

Target

  • Alternative Names
  • ENV; envelope protein; Endogenous retrovirus group K member 19 Env polyprotein

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.

Downloads

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Immunotoxin

CAT Product Name Application Type
AGTO-L012R anti-env immunotoxin 907 (IgG)-RTA Cytotoxicity assay, Functional assay

Single-domain Antibody

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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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