Anti-EGFR Recombinant Antibody (Imgatuzumab) (CAT#: TAB-753)

Recombinant humanized (from rat) antibody expressed in CHO binding to human EGFR. Imgatuzumab is a humanized monoclonal antibody designed for the treatment of cancer. It acts as an immunomodulator.

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Cyt

Figure 1 In vitro ADCC analysis of imgatuzumab, cetuximab or the combination.

PBMCs as effector cells were added to the untreated H441 (A), H292 (B) or A549 (C) target cells at a ratio of 25:1. ADCC responses were normalized to the ADCC response of the highest imgatuzumab concentration due to variations in responses between PBMCs of different donors. Absolute ADCC responses ranged between 12 and 53%. (*P < 0.05, **P < 0.01, ***P < 0.001 cetuximab vs imgatuzumab). (D) A549 cells were pretreated with imgatuzumab alone (20 µg/mL) or combined with cetuximab (20 µg/mL total) in culture medium for 72 hours. Next, cells with (open triangle and open square) or without (solid triangle and solid square) antibody pretreatment for 72 hours were washed and the ADCC assay was performed using PBMCs as effector cells and A549 cells as target cells at a ratio of 25:1. Data points are mean ± SD (n = 3).

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

Inhib

Figure 2 In vitro growth-inhibitory effect of anti-EGFR monoclonal antibody treatment.

Cells were seeded in 12-wells plates and allowed to adhere overnight. Next, H292, H322 and A549 cells were treated with the anti-EGFR monoclonal antibodies (20 µg/ mL total) for the indicated days under normal growth conditions without additional EGF (medium with 10% FCS) or in medium containing 10% FCS with 1 or 10 ng/mL EGF. Data points are mean + SD. All experiments were performed in triplicate.

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

WB

Figure 3 Cellular localization and endocytic trafficking of anti-EGFR monoclonal antibodies in SW-1573.

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

Inter

Figure 4 Effects of TKI treatment on monoclonal antibody-induced internalization and degradation.

(A) Western blot analysis of the effect of 24 hours anti-EGFR monoclonal antibody treatment (20 µg/mL total) with or without erlotinib (10 µM) on EGFR total protein levels in SW-1573, H292 and A549. Data points are mean + SD. (*P < 0.05, **P < 0.01, ***P < 0.001 compared to untreated control; $ P < 0.05, $ P < 0.01, $$ P < 0.001 compared to erlotinib treated control). (B) SW-1573 cells were pretreated for 24 hours with the EGFR TKI inhibitor erlotinib (10 µM). Cells were subsequently surface labeled with the anti-EGFR monoclonal antibodies (20 µg/mL total) on ice and incubated at 37 °C for 4 hours, and then analyzed for the non-internalized EGFR-antibody complexes (NI) and cell surface expression (SE). The lower the amount of non-internalized EGFR-antibody complexes, the higher the amount of internalized antibodyEGFR complexes. Surface expression of the control and erlotinib treated cells were both set at 100%. 24 hours erlotinib treatment resulted in a slight upregulation (10 – 15 % increase). Data points are mean ± SD (n = 3). (*P < 0.05).

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

Inter

Figure 5 Influence of monoclonal antibody binding on EGFR internalization and surface expression.

SW-1573 (A) and H292 cells (B) were surface labeled with the anti-EGFR monoclonal antibodies on ice and incubated at 37 °C for the indicated times, and then analyzed for the non-internalized EGFR-antibody complexes (left) and cell surface expression (right). The lower the amount of noninternalized EGFR-antibody complexes, the higher the amount of internalized antibody-EGFR complexes. The surface expression at t=0 was set at 100%. The non-internalized and surface expression were determined as described in Materials and Methods (n=3). Data points are mean ± SD. (**P < 0.01, ***P < 0.001 combination vs imgatuzumab; $ P < 0.01, $$ P < 0.001 combination vs cetuximab; two-way ANOVA followed by Bonferroni post-test).

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

WB

Figure 6 Western blot analysis of the effect of 24 hours anti-EGFR monoclonal antibody treatment on EGFR total protein levels and downstream signaling under normal growth conditions or 15 min EGF stimulation (10 ng/mL).

The immunoreactive spots were quantified by densitometric analysis and normalized using actin (see also Supplementary Figure 2). Values are expressed as fold increase versus control + SD. All experiments were performed in triplicate.

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

FC

Figure 7 Effect of anti-EGFR monoclonal antibody treatment on EGFR surface expression levels.

(A) Flow cytometric analysis of imgatuzumab and cetuximab binding alone or in combination in H322, SW-1573, H441, H292 and A549 cells. (B) H322, SW-1573, H292, H441 and A549 cells were treated with the anti-EGFR monoclonal antibodies (20 µg/mL total) for 72 hours. Surface expression levels were determined using flow cytometry. The surface expression in untreated control cells was set at 100% both for the single antibodies and the combination. (*P < 0.05, **P < 0.01 combination vs imgatuzumab; $ P < 0.05, $ P < 0.01, $$ P < 0.001 combination vs cetuximab; unpaired t-test). Data points are mean + SD (n = 3).

Kol, A., van Scheltinga, A. T., Pool, M., Gerdes, C., de Vries, E., & de Jong, S. (2017). ADCC responses and blocking of EGFR-mediated signaling and cell growth by combining the anti-EGFR antibodies imgatuzumab and cetuximab in NSCLC cells. Oncotarget, 8(28), 45432.

IHC

Figure 8 EGFR expression analysis and immunohistochemistry.

(A) Ex vivo tissue analysis. Mean vessel density (MVD), EGFR and hematoxylin and eosin immunohistochemical staining of A431, A549 and H441 tumors. (B) In vitro flow cytometric analysis of EGFR membrane expression in A431, A549 and H441 cells. (C) Ex vivo human EGFR extracellular domain levels in A431, A549 and H441 tumor tissue lysates. (D) EGFR histo-score (H-score) for A431, A549 and H441 tumors. (E) Mean vessel density score for A431, A549 and H441 tumors.

Pool, M., Kol, A., Lub-de Hooge, M. N., Gerdes, C. A., de Jong, S., de Vries, E. G., & van Scheltinga, A. G. T. (2016). Extracellular domain shedding influences specific tumor uptake and organ distribution of the EGFR PET tracer 89Zr-imgatuzumab. Oncotarget, 7(42), 68111.

ELISA

Figure 9 EGFR analysis of plasma, liver and cell lysates.

(A) Human EGFR extracellular domain concentration in plasma of A431, A549 and H441 xenograft bearing and non-tumor bearing control mice. (B) Human EGFR extracellular domain level in liver tissue lysates of A431, A549 and H441 xenograft bearing and non-tumor bearing control mice. (C) Human EGFR extracellular domain concentration in serum-free supernates of confluent cell cultures of A431, A549 and H441 cells. (D) Human EGFR extracellular domain concentration in serum-containing supernates of confluent cell cultures of A431, A549 and H441 cells, only statistical significances against A431 are shown for clarity.

Pool, M., Kol, A., Lub-de Hooge, M. N., Gerdes, C. A., de Jong, S., de Vries, E. G., & van Scheltinga, A. G. T. (2016). Extracellular domain shedding influences specific tumor uptake and organ distribution of the EGFR PET tracer 89Zr-imgatuzumab. Oncotarget, 7(42), 68111.


Specifications

  • Immunogen
  • The details of the immunogen for this antibody are not available.
  • Host Species
  • Rat
  • Derivation
  • Humanized (from rat)
  • Type
  • IgG1 - kappa
  • Species Reactivity
  • Human
  • Applications
  • Neut, ELISA, IF, IP, FuncS, FC, WB, Cyt, Inhib, IHC
  • CAS
  • 959963-46-3
  • Generic Name
  • imgatuzumab
  • MW
  • 145.0 kDa
  • Related Disease
  • Solid tumors

Applications

  • Application Notes
  • The EGFR antibody has been reported in applications of Neut, ELISA, IF, IP, FuncS, FC, WB, Cyt, Inhib, IHC.

Target

  • Alternative Names
  • imgatuzumab;959963-46-3;RG7160;GA201;RO5083945;ICR62;EGFR;epidermal growth factor receptor;epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) , ERBB;ERBB1;erythroblastic leukemia viral (v erb b) oncogene homo

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

  • Biosimilar Overview
  • Related Diseases
  • Related Signaling Pathways
Please click here for more details about the introduction to mechanism of action, clinical projects and approved drugs of Imgatuzumab.

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See other products for "Imgatuzumab"

Afuco™ Anti-EGFR ADCC Recombinant Antibody (Imgatuzumab), ADCC Enhanced
This product is an ADCC enhanced antibody produced by our Afuco™ platform. Recombinant humanized (from rat) antibody expressed in CHO binding to human EGFR. Imgatuzumab is a humanized monoclonal antibody designed for the treatment of cancer. It acts as an immunomodulator.

See other products for "EGFR"

Single-domain Antibody

CAT Product Name Application Type
NABG-056 Recombinant Anti-Mouse Egfr VHH Single Domain Antibody ELISA, IHC, FC, FuncS Llama VHH
PNBL-016 Recombinant Anti-Human EGFR VHH Single Domain Antibody (7D12) WB, ELISA Llama VHH
PNBL-017 Recombinant Anti-Human EGFR VHH Single Domain Antibody (9G8) FuncS, ELISA, IF Llama VHH
PNBL-018 Recombinant Anti-Human EGFR VHH Single Domain Antibody (EgA1) FuncS, SPR Llama VHH
HPAB-AP881-YC Recombinant Llama Anti-EGFR Single Domain Antibody (VHH122) ELISA, FC, IP, FuncS Llama VHH

Humanized Antibody

CAT Product Name Application Type
TAB-165 Anti-Human EGFR Recombinant Antibody (Matuzumab) Neut, ELISA, IF, IP, FuncS, FC, ICC IgG1
TAB-274MZ Human Anti-EGFR Recombinant Antibody (TAB-274MZ) FC Humanized IgG
TAB-274MZ-S(P) Human Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-274MZ-S(P)) FC Humanized scFv
TAB-274MZ-F(E) Human Anti-EGFR Recombinant Antibody; Fab Fragment (TAB-274MZ-F(E)) FC Humanized Fab
TAB-300MZ-F(E) Human Anti-EGFR Recombinant Antibody; Fab Fragment (TAB-300MZ-F(E)) ELISA Humanized Fab

Recombinant Antibody

CAT Product Name Application Type
PABZ-039 Mouse Anti-EGFR Recombinant Antibody (clone mAb528) FC Mouse IgG
PABW-039 Human Anti-EGFR Recombinant Antibody (clone Cetuximab (IMC-C225)) FuncS Chimeric (mouse/human) IgG
PABC-040 Human Anti-EGFR Recombinant Antibody (clone MR1) Block Human IgG
MOB-0094MZ Recombinant Mouse Anti-EGFR Antibody (clone FGF-R2) ELISA, IHC-P, FC Mouse antibody
FAMAB-0094YC Human Anti-EGFR Recombinant Antibody (clone F4) ELISA, WB, FC Human IgG

Fab Fragment Antibody

CAT Product Name Application Type
PFBZ-039 Mouse Anti-EGFR Recombinant Antibody (clone mAb528); Fab Fragment FC Mouse Fab
PFBW-039 Human Anti-EGFR Recombinant Antibody (clone Cetuximab (IMC-C225)); Fab Fragment FuncS Chimeric (mouse/human) Fab
PFBC-040 Human Anti-EGFR Recombinant Antibody (clone MR1); Fab Fragment Block Human Fab
PFBL-459 Human Anti-EGFR Recombinant Antibody (clone C225); Fab Fragment FC Human Fab
PFBL-460 Human Anti-EGFR Recombinant Antibody (clone h-R3); Fab Fragment ELISA, WB, FuncS Humanized Fab

Chimeric Antibody

CAT Product Name Application Type
TAB-272MZ Human Anti-EGFR Recombinant Antibody (TAB-272MZ) FuncS Chimeric (mouse/human) IgG
TAB-302MZ Human Anti-EGFR Recombinant Antibody (TAB-302MZ) ELISA Chimeric (mouse/human) IgG1
TAB-272MZ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-272MZ-S(P)) FuncS Mouse scFv
TAB-302MZ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-302MZ-S(P)) ELISA Mouse scFv
TAB-272MZ-F(E) Human Anti-EGFR Recombinant Antibody; Fab Fragment (TAB-272MZ-F(E)) FuncS Chimeric (mouse/human) Fab

Mouse Antibody

CAT Product Name Application Type
TAB-310MZ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-310MZ-S(P)) ELISA Mouse scFv
TAB-311MZ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-311MZ-S(P)) ELISA Mouse scFv
TAB-312MZ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-312MZ-S(P)) ELISA Mouse scFv
TAB-316MZ-S(P) Anti-Human EGFR Recombinant Antibody scFv Fragment (07D06) WB, MTT assay
TAB-317MZ-S(P) Anti-Human EGFR Recombinant Antibody scFv Fragment (12D03) WB, MTT assay

Human Antibody

CAT Product Name Application Type
TAB-313MZ-F(E) Anti-Human EGFR Recombinant Antibody Fab Fragment (EG-19-11) ELISA, FC Human antibody
TAB-314MZ-F(E) Anti-Human EGFR Recombinant Antibody Fab Fragment (EG-26-11) ELISA, FC Human antibody
TAB-315MZ-F(E) Anti-Human EGFR Recombinant Antibody Fab Fragment (528) WB, FC Human antibody
PABX-053 Recombinant Human Anti-EGFR Antibody (DL11) WB, ELISA, FuncS IgG
PABX-053-F (E) Recombinant Human Anti-EGFR Antibody Fab Fragment (DL11) WB, ELISA, FuncS Fab

Fc Glycosylation

CAT Product Name Application Type
Gly-055LC Recombinant Anti-Human EGFR Antibody (Fc glycosylation/High-mannose glycosylated) ELISA Chimeric antibody (mouse/human)
Gly-144LC Recombinant Anti-Human EGFR Antibody (Fc glycosylation) ELISA Humanized antibody

High-mannose Glycoform

CAT Product Name Application Type
Gly-055LC-1 Recombinant Anti-Human EGFR Antibody (Fc glycosylation/High-mannose glycosylated) ELISA Chimeric antibody (mouse/human)

Deglycosylated Antibody (Non-glycosylated IgGs)

CAT Product Name Application Type
Gly-167LC Recombinant Anti-Human EGFR Antibody (Non-glycosylated) ELISA Human antibody

Chicken IgY Antibody

CAT Product Name Application Type
BRD-0183MZ Chicken Anti-EGFR Polyclonal IgY WB Chicken antibody
BRD-0668MZ Chicken Anti-EGFR Polyclonal IgY WB Chicken antibody

MHC Tetramer for Cancer

CAT Product Name Application Type
MHC-LC773 A*0201/Human EGFR (YLNTVQPTCV) MHC Tetramer FCM
MHC-LC4545 PE-DQB1*03:02/Human EGFR (SRALEEKKGNYVVTHG) MHC Tetramer FCM
MHC-YF409 A*02:01/Human EGF-R-479 (KLFGTSGQKT) MHC Pentamer FCM

Neutralizing Antibody

CAT Product Name Application Type
NEUT-722CQ Rabbit Anti-EGFR Recombinant Antibody (clone CBL1011) Neut Rabbit IgG
NEUT-723CQ Mouse Anti-EGFR Recombinant Antibody (clone CBL931) WB, IP, IHC, ICC, Neut Mouse IgG1
NEUT-724CQ Rabbit Anti-EGFR Recombinant Antibody (clone D1D4J) IF, FC, WB, IP, Neut Rabbit IgG

Rabbit Monoclonal Antibody

CAT Product Name Application Type
MOR-1101 Hi-Affi™ Rabbit Anti-EGFR Recombinant Antibody (clone DS1101AB) IHC-P Rabbit IgG
MOR-4520 Hi-Affi™ Rabbit Anti-EGFR Recombinant Antibody (clone TH28DS) IF, ICC, FC Rabbit IgG
MOR-4570 Hi-Affi™ Rabbit Anti-EGFR Recombinant Antibody (clone TH82DS) ELISA Rabbit IgG
MOR-4571 Hi-Affi™ Rabbit Anti-EGFR Recombinant Antibody (clone TH83DS) WB, IF, ICC, FC Rabbit IgG
MOR-0033-FY Rabbit Anti-EGFR Recombinant Antibody (clone AFY0004) ICC, IHC, WB Rabbit IgG

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-165 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Matuzumab), ADCC Enhanced Neut, ELISA, IF, IP, FuncS, FC ADCC enhanced antibody
AFC-TAB-464CQ Afuco™ Anti-EGFR ADCC Recombinant Antibody (Tomuzotuximab), ADCC Enhanced ELISA, IHC, FC, IP, IF, FuncS ADCC enhanced antibody
AFC-TAB-003 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Cetuximab), ADCC Enhanced IF, IP, Neut, FuncS, ELISA, FC ADCC enhanced antibody
AFC-TAB-040 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Zalutumumab), ADCC Enhanced ELISA, FC, IP, FuncS, IF, Neut ADCC enhanced antibody
AFC-TAB-119 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Necitumumab), ADCC Enhanced FC, IP, ELISA, Neut, FuncS, IF ADCC enhanced antibody

scFv Fragment Antibody

CAT Product Name Application Type
HPAB-S0021-YC-S(P) Human Anti-EGFR Recombinant Antibody (clone E6.3); scFv Fragment ELISA, Inhib Human scFv
HPAB-2059-FY-S(P) Mouse Anti-EGFR Recombinant Antibody (clone L2-12B); scFv Fragment FC, ELISA Mouse scFv
HPAB-2060-FY-S(P) Mouse Anti-EGFR Recombinant Antibody (clone L3-11D); scFv Fragment FC, ELISA Mouse scFv
HPAB-1530WJ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (clone 340) ELISA, WB Mouse scFv
HPAB-1531WJ-S(P) Human Anti-EGFR Recombinant Antibody; scFv Fragment (clone hu340) ELISA, WB Human scFv

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* 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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