Recombinant Anti-Human EGFR VHH Single Domain Antibody (7D12) (CAT#: PNBL-016)

Anti-Human EGFR VHH Single Domain Antibody is a recombinant protein produced in E. coli.

Host: Llama

Isotype: Llama VHH

Applications: WB, ELISA


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Figure 1 In vitro characterisation of selected monovalent anti‐EGFR sdAbs.

The binding of biotinylated EGF (800 pM) to an EGFR‐ECD‐Fc fusion was tested in the presence of increasing amounts of the 9G8, 7D12, 7C12 or 38G7 sdAbs, or without sdAb. Receptor‐bound EGF was then detected via peroxidase‐coupled streptavidin and staining with OPD/H2O2.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

Figure 2 In vitro characterisation of selected monovalent anti‐EGFR sdAbs.

The binding of 7D12 sdAbs to EGFR, expressed on phage, was detected in a 100-fold molar excess of the indicated antibody (cetuximab) or antibody fragments.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

Inhib

Figure 3 Inhibition of A431 tumour cell proliferation by monovalent and bivalent sdAbs.

The proliferation of A431 cells in the presence of increasing amounts of sdAb monovalent and bivalent 7D12; was measured using a sulphorhodaminebased stain of total cellular protein after TCA precipitation. Proliferation is plotted as percentage of maximal growth (cells left without treatment). The whole antibody cetuximab was used as ''standard'' in every test. Data points where the value of the control is statistically different from that of the sdAb-treated group are indicated by an asterisk.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

Inhib

Figure 4 Inhibition of A431 tumour cell proliferation by monovalent and bivalent sdAbs.

The proliferation of A431 cells in the presence of increasing amounts of sdAb mixtures of monovalent and bivalent 7D12 and 9G8; was measured using a sulphorhodaminebased stain of total cellular protein after TCA precipitation. Proliferation is plotted as percentage of maximal growth (cells left without treatment). The whole antibody cetuximab was used as ''standard'' in every test. Data points where the value of the control is statistically different from that of the sdAb-treated group are indicated by an asterisk; only different between 7D12þ9G8 and control.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

FuncS

Figure 6 In vitro optimalisation and characterisation of trivalent biparatopic sdAb CONAN-1.

The proliferation of A431 cells in the presence of increasing amounts of biparatopic sdAb 7D12-9G8 (with linkers varying in length between the two sdAb units) was measured using a sulphorhodamine-based stain of total cellular protein after TCA precipitation. Proliferation is plotted as percentage of maximal growth (cells left without treatment). Error bars have been omitted to increase the clarity of the figure. The different linker lengths between the two sdAbs are indicated.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

FuncS

Figure 7 In vitro optimalisation and characterisation of trivalent biparatopic sdAb CONAN-1.

Purified EGFR-ECD-Fc fusion protein was immobilised via a coated anti-human Fc anti-serum, and a concentration range of either bivalent (7D12-9G8) or trivalent (CONAN-1) sdAb was bound. The binding of biotinylated mouse serum albumin (MSA) was subsequently detected with peroxidase-conjugated streptavidin and staining with OPD/H2O2.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

FuncS

Figure 8 In vitro optimalisation and characterisation of trivalent biparatopic sdAb CONAN-1.

The proliferation of A431 cells in the presence of increasing amounts of sdAb was measured using the sulphorhodamine B assay. The effect of the presence of human serum albumin (HSA: 1%) in the medium was assessed.

Roovers, R. C., Vosjan, M. J., Laeremans, T., el Khoulati, R., de Bruin, R. C., Ferguson, K. M., ... & van Bergen en Henegouwen, P. M. (2011). A biparatopic anti‐EGFR sdAb efficiently inhibits solid tumour growth. International journal of cancer, 129(8), 2013-2024.

FuncS

Figure 9 In vitro specificity of ⁹⁹ᵐTc-7C12 and ⁹⁹ᵐTc-7D12 on A431 cell line after 2-h incubation on ice.

Binding of labeled compound was blocked by 1,000-fold excess of unlabeled 99mTc-7C12, ⁹⁹ᵐTc-7D12, or cetuximab.

Gainkam, L. O. T., Huang, L., Caveliers, V., Keyaerts, M., Hernot, S., Vaneycken, I., ... & Lahoutte, T. (2008). Comparison of the biodistribution and tumor targeting of two 99mTc-labeled anti-EGFR sdAbs in mice, using pinhole SPECT/micro-CT. Journal of Nuclear Medicine, 49(5), 788-795.

FuncS

Figure 10 Tumor uptake (%IA/g) of ⁹⁹ᵐTc-7D12 expressed as function of tumor weight in nu/nu mouse A431 xenografts.

Animals were dissected at 1.5 h after injection of tracer. Tumor uptake, in %IA/g, decreased with increasing tumor size.

Gainkam, L. O. T., Huang, L., Caveliers, V., Keyaerts, M., Hernot, S., Vaneycken, I., ... & Lahoutte, T. (2008). Comparison of the biodistribution and tumor targeting of two 99mTc-labeled anti-EGFR sdAbs in mice, using pinhole SPECT/micro-CT. Journal of Nuclear Medicine, 49(5), 788-795.

FuncS

Figure 11 Blood clearance.

Non–tumor-bearing nu/nu mice (n53) received intravenous injection of ⁹⁹ᵐTc-7C12 or 99mTc7D12. Blood samples were collected at indicated time points and analyzed with g-counter. Half-life was determined using biexponential nonlinear regression fit (GraphPad Prism).

Gainkam, L. O. T., Huang, L., Caveliers, V., Keyaerts, M., Hernot, S., Vaneycken, I., ... & Lahoutte, T. (2008). Comparison of the biodistribution and tumor targeting of two 99mTc-labeled anti-EGFR sdAbs in mice, using pinhole SPECT/micro-CT. Journal of Nuclear Medicine, 49(5), 788-795.

Figure 12 7D12 and 425scFv retain binding capacity to EGFR after coupling to liposomes and EGFR downregulation is a specific feature of sdAb-liposomes.

The binding of 7D12 to EGFR was tested in the presence of: EGa1 sdAb (EGa1), EGa1-liposomes (EGa1-L 0.4 and EGa1-L 0.8), 7D12 sdAb (7D12), 7D12- liposomes (7D12-L 0.4), 425 single chain variable fragment (425scFv), 425scFvliposomes (425scFv-L 0.4), Erbitux, and liposomes (L). Bound phage were detected with a HRP-coupled anti-M13 monoclonal antibody and staining with OPD/H2O.

Oliveira, S., Schiffelers, R. M., van der Veeken, J., van der Meel, R., Vongpromek, R., en Henegouwen, P. M. V. B., ... & Roovers, R. C. (2010). Downregulation of EGFR by a novel multivalent sdAb-liposome platform. Journal of Controlled Release, 145(2), 165-175.

Figure 13 7D12 and 425scFv retain binding capacity to EGFR after coupling to liposomes and EGFR downregulation is a specific feature of sdAb-liposomes.

The binding of EGa1-phage to EGFR was tested in the presence of: EGa1 sdAb (EGa1), EGa1-liposomes (EGa1-L 0.4 and EGa1-L 0.8), 7D12 sdAb (7D12), 7D12- liposomes (7D12-L 0.4), 425 single chain variable fragment (425scFv), 425scFvliposomes (425scFv-L 0.4), Erbitux, and liposomes (L). Bound phage were detected with a HRP-coupled anti-M13 monoclonal antibody and staining with OPD/H2O.

Oliveira, S., Schiffelers, R. M., van der Veeken, J., van der Meel, R., Vongpromek, R., en Henegouwen, P. M. V. B., ... & Roovers, R. C. (2010). Downregulation of EGFR by a novel multivalent sdAb-liposome platform. Journal of Controlled Release, 145(2), 165-175.

Figure 14 7D12 and 425scFv retain binding capacity to EGFR after coupling to liposomes and EGFR downregulation is a specific feature of sdAb-liposomes.

Cells were incubated for 4 h at 37 °C with different liposome formulations: EGa1-L, 7D12-L and 425scFv-L. After the incubation with liposomes at 0.5, 0.25 and 0.125 mM TL concentrations, cells were further incubated for 2 days. Thereafter, the level of cell surface EGFR was determined.

Oliveira, S., Schiffelers, R. M., van der Veeken, J., van der Meel, R., Vongpromek, R., en Henegouwen, P. M. V. B., ... & Roovers, R. C. (2010). Downregulation of EGFR by a novel multivalent sdAb-liposome platform. Journal of Controlled Release, 145(2), 165-175.


Specifications

  • Immunogen
  • Human epidermal growth factor receptor
  • Host Species
  • Llama
  • Derivation
  • Llama
  • Type
  • Llama VHH
  • Species Reactivity
  • Human
  • Clone
  • 7D12
  • Applications
  • WB, ELISA

Product Property

  • Purity
  • >95% by SDS-PAGE and HPLC analysis
  • Storage
  • Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing of samples.

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

Related Resources

  • Related Diseases
  • Related Signaling Pathways

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.

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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
TAB-015MZ-VHH Anti-Human EGFR Recombinant Antibody (PMP7A5) sELISA Single domain antibody
TAB-016MZ-VHH Anti-Human EGFR Recombinant Antibody (PMP7D12) sELISA Single domain antibody
HPAB-1027WJ Recombinant Camelid Anti-EGFR Single Domain Antibody ELISA, WB Camelid VHH
HPAB-0705-YJ-VHH Camelid Anti-EGFR Recombinant Single Domain Antibody (EG2) ELISA, In vivo staining Camelid 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-710 Anti-EGFR Recombinant Antibody (Nimotuzumab) ELISA, IP, FC, FuncS, Neut, IF, IHC IgG1 - kappa
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

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
PSBW-088 Mouse Anti-EGFR Recombinant Antibody (clone 7A7); scFv Fragment ELISA, WB Mouse scFv
FAMAB-0094YC Human Anti-EGFR Recombinant Antibody (clone F4) ELISA, WB, FC Human IgG

Chimeric Antibody

CAT Product Name Application Type
TAB-0225CL-S(P) Human Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-0225CL-S(P)) Block, Inhib, FuncS, Apop, In vivo Human scFv
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

Mouse Antibody

CAT Product Name Application Type
TAB-0565CL-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-0565CL-S(P)) ELISA Mouse scFv
TAB-307MZ-S(P) Mouse Anti-EGFR Recombinant Antibody; scFv Fragment (TAB-307MZ-S(P)) ELISA, FC Mouse scFv
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

Human Antibody

CAT Product Name Application Type
TAB-313MZ Anti-Human EGFR Recombinant Antibody (EG-19-11) ELISA, FC Human antibody
TAB-314MZ Anti-Human EGFR Recombinant Antibody (EG-26-11) ELISA Human antibody
TAB-299MZ-F(E) Human Anti-EGFR Recombinant Antibody; Fab Fragment (TAB-299MZ-F(E)) Internalization Assay Human Fab
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-4675 Hi-Affi™ Rabbit Anti-EGFR Recombinant Antibody (clone TH189DS) WB, IF, ICC, FC Rabbit IgG
MOR-4676 Hi-Affi™ Rabbit Anti-EGFR Recombinant Antibody (clone TH190DS) WB, IF, ICC, FC Rabbit IgG
MOR-0033-FY Rabbit Anti-EGFR Recombinant Antibody (clone AFY0004) ICC, IHC, WB Rabbit IgG

Fab Fragment Antibody

CAT Product Name Application Type
HPAB-0010-YC-F(E) Mouse Anti-EGFR Recombinant Antibody (clone L211C); Fab Fragment ELISA, FC Mouse Fab
HPAB-0011-YC-F(E) Human Anti-EGFR Recombinant Antibody (clone Pep 1); Fab Fragment FC Human Fab
HPAB-0012-YC-F(E) Human Anti-EGFR Recombinant Antibody (clone Pep 2); Fab Fragment FC Human Fab
HPAB-0013-YC-F(E) Human Anti-EGFR Recombinant Antibody (clone Pep 3); Fab Fragment FC Human Fab
HPAB-0014-YC-F(E) Human Anti-EGFR Recombinant Antibody (clone Pep 4); Fab Fragment FC Human Fab

scFv Fragment Antibody

CAT Product Name Application Type
FAMAB-0094-YC-S(P) Human Anti-EGFR Recombinant Antibody (clone F4); scFv Fragment ELISA, WB, FC Human 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
HPAB-1563WJ-S(P) Human Anti-EGFR Recombinant Antibody; scFv Fragment (clone chMint5) ELISA, WB Human scFv

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-710 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Nimotuzumab), ADCC Enhanced ELISA, IP, FC, FuncS, Neut, IF ADCC enhanced antibody
AFC-TAB-753 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Imgatuzumab), ADCC Enhanced Neut, ELISA, IF, IP, FuncS, FC ADCC enhanced antibody
AFC-TAB-H35 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Futuximab), ADCC Enhanced FuncS, IF, Neut, ELISA, FC, IP ADCC enhanced antibody
AFC-TAB-H49 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Modotuximab), ADCC Enhanced FuncS, IF, Neut, ELISA, FC, IP ADCC enhanced antibody
AFC-TAB-020 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Panitumumab), ADCC Enhanced ELISA, IP, FC, FuncS, Neut, IF ADCC enhanced antibody

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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.

* 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
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
WBWestern Blot

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