Anti-Human EGFR Therapeutic Antibody (mAb806 ) (CAT#: TAB-326MZ)

This antibody can recognize the unique truncation mutant, EGFRvIII. In xenograft models, mAb806 has exhibited a potent anti-tumor activity with no targeting of normal tissues.

  • Published Data
  • Datasheet
  • MSDS
  • COA
FuncS

Figure 1 Effect of mAb 806 treatment on tumour angiogenesis.

(a) Bar graphs showing the mean change in H-score between mAb 806-treated and control tumours for the biological parameters shown. (b) Representative (250×) images of CD31+ blood vessels in the mAb 806-treated or control tumours, stratified by intratumoral position. scale bar: 25 μM. (c) Bar graphs showing the mean change in the MVD, MSA and total vessel area in the mAb 806-treated or control tumours.

Gan, H. K., Lappas, M., Cao, D. X., Cvrljevdic, A., Scott, A. M., & Johns, T. G. (2009). Targeting a unique EGFR epitope with monoclonal antibody 806 activates NF‐κB and initiates tumour vascular normalization. Journal of cellular and molecular medicine, 13(9b), 3993-4001.

FuncS

Figure 2 Effect of mAb 806 treatment on blood vessel function.

(a) Blood vessels with continuous pericyte coverage (arrowhead) or incomplete pericyte coverage (arrow), scale bar: 20 μM. Bar graphs showing the mean change in the density of (b) vessels with any pericyte coverage and (c) vessels with complete pericyte coverage in the mAb 806-treated or control tumours. Bar graphs showing the effects of mAb 806 treatment on (d) the mean oxygenation at day 17, (e) the prevalence of severe hypoxia (less than 2.5 mmHg) and (f) the mean oxygenation in case-control analysis (i.e. matched for tumour volume).

Gan, H. K., Lappas, M., Cao, D. X., Cvrljevdic, A., Scott, A. M., & Johns, T. G. (2009). Targeting a unique EGFR epitope with monoclonal antibody 806 activates NF‐κB and initiates tumour vascular normalization. Journal of cellular and molecular medicine, 13(9b), 3993-4001.

FuncS

Figure 3 In vivo function of NF-κB and VEGF on mAb 806 anti-tumour activity.

Treatment with mAb 806 significantly increases nuclear NF-κB DNA (a) as measured by ELISA or (b) Western blot of the p65 subunit. (c) Survival curves using a combined end-point of moribund condition or tumour volume reaching 1000 mm³ in mice treated with vehicle, mAb 806 (1 mg), BAY 11-7085 (100 μg) or a combination of both on the days indicated. (d) Survival curves using a combined end-point of moribund condition or tumour volume reaching 1500 mm³ in mice treated with vehicle, mAb 806 (1 mg), bevacizumab (400 μg) or a combination of both on the days indicated.

Gan, H. K., Lappas, M., Cao, D. X., Cvrljevdic, A., Scott, A. M., & Johns, T. G. (2009). Targeting a unique EGFR epitope with monoclonal antibody 806 activates NF‐κB and initiates tumour vascular normalization. Journal of cellular and molecular medicine, 13(9b), 3993-4001.

FC

Figure 4 Flow cytometric analysis of BaF/3, A431, and U87MG.Δ2-7 cell lines

Cells were incubated with either an irrelevant IgG2b, mAb 806, or mAb 528 following conjugation with Cy3 dye. (A-C) BaF/3 cells were directly labeled with IgG2b/Cy3, mAb 806/Cy3, and mAb 528/Cy3, respectively; binding was visualized by flow cytometry. (D and E) A431 cells were directly labeled with IgG2b/Cy3, mAb 806/Cy3, and mAb 528/Cy3, respectively; binding was visualized by flow cytometry. (G and H) U87MG.Δ2-7 cells were directly labeled with IgG2b/Cy3, mAb 806/Cy3, and mAb 528/Cy3, respectively; binding was visualized by flow cytometry.

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.

Figure 5 Time-lapse microscopy of antibody internalization in A431 cells.

(A) Cells were preincubated with mAb 806-Cy3 at 4°C (0 minute). Internalized antibody is indicated by arrows in sequential images representing 15, 30, and 60 minutes of incubation at 37°C. (B) Mean ratio of fluorescence intensity between the intracellular region and cell surface. The mean fluorescence of the intracellular region (dashed circle) and cell membrane (dashed rectangle) were calculated (n = 5) and the average ratio of intracellular to membrane (black plot) and membrane to intracellular (red plot) fluorescence ± SEM plotted over time (n = 5).

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.

Figure 6 Time-lapse of mAb 806 internalization in A431 cells expressing dynamin K44A-GFP.

(A) Cells were transiently transfected with DynK44A-GFP (red asterisk). Panels representing images at 5, 10, 20, 30, and 60 minutes after mAb 806-Cy3 internalization are shown. (B) Mean ratio of intracellular to membrane (black) and membrane to intracellular (red) fluorescence of untransfected wt cells ± SEM (n = 4), over time. Untransfected cells show gradual increase in intracellular fluorescence with a concurrent decrease in membrane associated fluorescence. (C) Mean ratio of intracellular to membrane (black) and membrane to intracellular (red) fluorescence of DynK44A-transfected cells ± SEM (n = 6), over time. DynK44A-transfected cells show little accumulation of intracellular fluorescence while sustaining high membrane fluorescence intensity.

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.

Figure 7 Time-lapse microscopy of internalization and colocalization of mAb 806-Cy3 (red) and transferrin-FITC (green) in A431 cells.

(A) Internalization of mAb 806 at 2 minutes is represented by yellow arrowheads. Colocalization is indicated by white arrows. Scale bar, 20 µm. Refer to Supplemental data for Video W3. (B) Colocalization of mAb 806-Cy3 (red) and EEA1 (green) in A431 cells. Following internalization of mAb 806, cells were fixed at different time intervals and stained for EEA1 localization. Colocalization is indicated by arrows in the merged images. Scale bar, 20 µm.

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.

Figure 8 Lysosomal localization of mAb 806-Cy3 in A431 Cells.

(A) Preincubated mAb 806-Cy3 (red) was allowed to internalize for 60 minutes at 37°C. Cells were subsequently fixed and stained for LAMP1 (green). Colocalization is indicated by arrows in the merged images. Scale bar, 20 µm. (B) Time-lapse microscopy of internalization and colocalization of mAb 806-Cy3 (red) and lgp-120-GFP (green) in A341 cells. Sequential images of transfected A431 cells are shown at 0, 30, 45, and 60 minutes. Colocalization is indicated by the arrows.

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.

Figure 9 Electron microscopic analysis of mAb 806 internalization and intracellular trafficking following binding to amplified EGFR in A431 cells.

(A-B) Gold particles (arrowheads) were detected in deeply invaginated clathrincoated pits after 5 minutes at 37°C. (C) Gold particles were also localized in free clathrin-coated vesicles within the cytoplasm. (D) No gold particles were present in structures resembling caveolae (open arrowheads) at 5 minutes. (E) Gold particles were also detected in tubular-vesicular structures resembling early endosomes after 10 to 15 minutes at 37°C. (F) After 30 minutes, gold particles were localized to multivesicular bodies.

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.

Figure 10 Biodistribution of mAb 806.

(A) Biodistribution (% ID/g) of ̶¹¹¹In-CHX-A″-DTPA- (closed symbols) and ̶¹²⁵I- (open symbols) labeled mAb 806 in A431 tumor xenografts and blood from BALB/c nude mice. Twenty mice received IV injections of a total of 10 µg (10 µCi) of radiolabeled mAb 806. Means for each time point consisting of data from four mice are shown. Bars, SD. Data are presented as: ̶¹¹¹In-blood (●); ̶¹¹¹In-A431 tumor (■); ¹²⁵I-blood (○); ̶¹²⁵I-A431 tumor (□). (B) Biodistribution of ̶¹²⁵I- (white bar) and ̶¹¹¹In-CHX-A″-DTPA- (black bar) labeled mAb 806 in normal murine tissues, A431 tumors, and blood at 4, 24, 48, 72, and 168 hours after IV administration (10 µCi, 10 µg total mAb 806) of BALB/c nude mice. The biodistribution of ̶¹²⁵I- (white bar) and ¹¹¹In-CHX-A″-DTPA- (black bar) labeled control mAb 100-310 was also assessed at 24 hours postinjection.

Perera, R. M., Zoncu, R., Johns, T. G., Pypaert, M., Lee, F. T., Mellman, I.,... & Scott, A. M. (2007). Internalization, intracellular trafficking, biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody.Neoplasia, 9(12), 1099-1110.


Specifications

  • Immunogen
  • Cells expressing EGFR vIII mutant
  • Host Species
  • Mouse
  • Specificity
  • Human
  • Clone
  • mAb806
  • Applications
  • FC, IF, FuncS
  • Related Disease
  • Glioblastoma

Applications

  • Application Notes
  • The antibody was validated for Flow Cytometry, Immunofluorescence, Function Assay. For details, refer to Published Data.

Target

  • Alternative Names
  • EGFR; epidermal growth factor receptor; ERBB; HER1; mENA; ERBB1; PIG61; NISBD2; proto-oncogene c-ErbB-1; cell growth inhibiting protein 40; erb-b2 receptor tyrosine kinase 1; cell proliferation-inducing protein 61; receptor tyrosine-protein kinase erbB-1; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog

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.

See other products for "EGFR"

* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

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