Chimeric (mouse/Human) Anti-TNFRSF8 Recombinant Antibody (clone Brentuximab) (CAT#: TAB-153)

This product is a human IgG1, κ antibody that can recognize human TNFRSF8.

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Block

Figure 1 Brentuximab vedotin blocks proliferation of PEL cells.

Human PEL cell lines BC-1 (A), BC-3 (B), UM-PEL-1c (C), and UM-PEL-3c (D) were treated with brentuximab vedotin (B.V.) at indicated doses for 0, 24, 48, and 72 hours. Proliferative response at each time point was measured by MTS assay.

Bhatt, S., Ashlock, B. M., Natkunam, Y., Sujoy, V., Chapman, J. R., Ramos, J. C.,... & Lossos, I. S. (2013). CD30 targeting with brentuximab vedotin: a novel therapeutic approach to primary effusion lymphoma. Blood, 122(7), 1233-1242.

Inhib

Figure 2 Rentuximab vedotin induces G2/M cell cycle arrest of PEL cells.

PEL cell lines BC-1 (A), BC-3 (B), UM-PEL-1c (C), and UM-PEL-3c (D) were treated with brentuximab vedotin (B.V.) at increasing concentrations. At 24 hours after treatment, cells were stained with propidium iodide to measure DNA content and analyzed by flow cytometry for cell cycle distribution. Bar graphs indicate the percentage of cells in different phases of cell cycle (G0, G1, S, G2/M).

Bhatt, S., Ashlock, B. M., Natkunam, Y., Sujoy, V., Chapman, J. R., Ramos, J. C.,... & Lossos, I. S. (2013). CD30 targeting with brentuximab vedotin: a novel therapeutic approach to primary effusion lymphoma. Blood, 122(7), 1233-1242.

Cyt

Figure 3 Brentuximab vedotin triggers apoptosis of PEL cells.

Lymphoma cell lines lacking CD30 expression WSU-NHL (A) and Raji (B) and CD30-expressing PEL cell lines BC-1 (C), BC-3 (D), UM-PEL-1c (E), and UM-PEL-3c (F) were treated with increasing concentrations of brentuximab vedotin (B.V.) or Ig-VcMMAE. At 72 hours after treatment, cell viability was determined by flow cytometry following YO-PRO and propidium iodide staining.

Bhatt, S., Ashlock, B. M., Natkunam, Y., Sujoy, V., Chapman, J. R., Ramos, J. C.,... & Lossos, I. S. (2013). CD30 targeting with brentuximab vedotin: a novel therapeutic approach to primary effusion lymphoma. Blood, 122(7), 1233-1242.

ELISA

Figure 4 CD30 levels, CD30 internalization, and brentuximab vedotin cell surface binding.

(A) sCD30 levels measured by ELISA in indicated cell lines and in cells directly derived from mice ascites. Error bar represents standard error of the mean between triplicate wells. (B) Cell surface CD30 expression and (C) cell surface brentuximab vedotin binding. BC-3, UM-PEL-1c, UM-PEL-3c, and Karpas 299 cells were incubated with 15 μg/mL brentuximab vedotin for 0, 24, and 48 hours followed by incubation with anti-CD30 (fluorescein isothiocyanate [FITC]) (B) or anti-hIgG (FITC) (C) to determine antigen-binding capacity values for CD30 (B) and of bound brentuximab vedotin (C), respectively.

Bhatt, S., Ashlock, B. M., Natkunam, Y., Sujoy, V., Chapman, J. R., Ramos, J. C.,... & Lossos, I. S. (2013). CD30 targeting with brentuximab vedotin: a novel therapeutic approach to primary effusion lymphoma. Blood, 122(7), 1233-1242.

Activ

Figure 5 Brentuximab vedotin extends the survival of PEL xenograft mice.

Kaplan-Meier survival curves of PEL xenograft mice. NOD/SCID mice (n = 5/group) were injected with 25 × 106 UM-PEL-1 (A) and UM-PEL-3 (B) cells. At 3 days postinjection, mice were treated for 3 weeks with interperitoneal injections of brentuximab vedotin (B.V.), PBS, or isotype-matched irrelevant Ig-vcMMAE.

Bhatt, S., Ashlock, B. M., Natkunam, Y., Sujoy, V., Chapman, J. R., Ramos, J. C.,... & Lossos, I. S. (2013). CD30 targeting with brentuximab vedotin: a novel therapeutic approach to primary effusion lymphoma. Blood, 122(7), 1233-1242.

FC

Figure 6 Brentuximab vedotin reduces viable cell number in CD30-positive but not CD30-negative GCT cells.

The CD30-positive EC cell lines GCT27 and NCCIT as well as the CD30-negative choriocarcinoma line JAR were treated with MMAE (A) or brentuximab vedotin (B-D). GCT27 (B), JAR (C) and NCCIT (D) were exposed to 250 ng/ml brentuximab vedotin. After 24, 48, 72 and 96 hrs of culture, cells were resuspended in equal volume for analysis. Viable Hoechst-negative cells were enumerated for 180 sec. by flow cytometry and are represented as multiples (x-fold) of the untreated control obtained at 24 hrs. To further evaluate dose-dependent effects to brentuximab vedotin, the three cell lines were exposed for 96 hrs to 250, 500 and 1000 ng/ml of the ADC as well as 100 pM MMAE (E). Enumerated viable Hoechst-negative cells are expressed in percent of the untreated control at 96 hrs.

Götz, B., van Beekum, C., Nettersheim, D., Schorle, H., Calaminus, G., Leuschner, I.,... & Schönberger, S. (2015). brentuximab Vedotin Presents Profound Anti-tumor Efficacy In Cd30+ And Co-cultured Cd30-Germ Tumor Cells: o-129. Pediatric Blood & Cancer, 62, S179.

Activ

Figure 7 Brentuximab vedotin exerts pronounced bystander activity on MMAE-sensitive, CD30-negative GCT cells in coculture with CD30-positive embryonal carcinoma.

For determination of bystander efficacy after drug exposure, cells were stained with CSFE and anti-CD30.PE (BER-H2, eBiosience/Germany). After 96 hrs of drug exposure, cell cultures were resuspended in equal volume and acquired for 180 sec. by flow cytometry. CD30-negative JAR cells are EPCAM positive (y-axis) but CD30 negative (x-axis) while GCT27 cells are EPCAM and CD30 positive. Hoechst-negative viable CD30-positive and CD30-negative subpopulations were assessed separately after gating on the respective cell fraction (A). Viable cell numbers are expressed in per cent of untreated control (B). Proliferation is investigated by CSFE dilution upon cellular division. To indicate inhibition of proliferation MFI of experimental conditions was normalized to the MFI of untreated cells and presented as x-fold MFI (C). Cell death was quantified as the proportion of Hoechstpositive cells of the entirety of acquired cells (D)

Götz, B., van Beekum, C., Nettersheim, D., Schorle, H., Calaminus, G., Leuschner, I.,... & Schönberger, S. (2015). brentuximab Vedotin Presents Profound Anti-tumor Efficacy In Cd30+ And Co-cultured Cd30-Germ Tumor Cells: o-129. Pediatric Blood & Cancer, 62, S179.


Specifications

  • Immunogen
  • Hodgkin"s lymphoma cell line L428.
  • Host Species
  • Human
  • Derivation
  • Chimeric (mouse/Human)
  • Type
  • Human IgG1, κ
  • Specificity
  • Human TNFRSF8
  • Species Reactivity
  • Human
  • Clone
  • Brentuximab
  • Applications
  • Neut, ELISA, IF, IP, FuncS, FC, ICC

Applications

  • Application Notes
  • This antibody has been reported for use in Neutralization, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Immunoprecipitation, Functional Assay, Flow Cytometry, Immunocytochemistry.

Target

  • Alternative Names
  • CD30; Ki-1; D1S166E; tumor necrosis factor receptor superfamily member 8; CD30L receptor; Ki-1 antigen; cytokine receptor CD30; lymphocyte activation antigen CD30

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.

Related Resources

Please click here for more details about the introduction to mechanism of action, clinical projects and approved drugs of Brentuximab vedotin.

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For Research Use Only. Not For Clinical Use.

* 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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