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.

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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

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.

Related Resources

  • Related Diseases
  • Related Signaling Pathways

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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
HPAB-AP881-YC Recombinant Llama Anti-EGFR Single Domain Antibody (VHH122) ELISA, FC, IP, FuncS Llama VHH
HPAB-1844-FY Camelid Anti-EGFR Recombinant Single Domain Antibody (clone 1.33) ELISA Camelid VHH
HPAB-1845-FY Camelid Anti-EGFR Recombinant Single Domain Antibody (clone 1.4) ELISA Camelid VHH

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
PABL-462 Human Anti-EGFR Recombinant Antibody (clone C225) FC Human IgG
PABL-463 Human Anti-EGFR Recombinant Antibody (clone h-R3) ELISA, WB, FuncS Humanized 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

Mouse Antibody

CAT Product Name Application Type
TAB-0564CL Mouse Anti-EGFR Recombinant Antibody (TAB-0564CL) ELISA Mouse IgG
TAB-0565CL Mouse Anti-EGFR Recombinant Antibody (TAB-0565CL) ELISA Mouse IgG
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-301MZ-F(E) Mouse Anti-EGFR Recombinant Antibody; Fab Fragment (TAB-301MZ-F(E)) ELISA Mouse Fab

Chimeric Antibody

CAT Product Name Application Type
TAB-0225CL-F(E) Human Anti-EGFR Recombinant Antibody; Fab Fragment (TAB-0225CL-F(E)) Block, Inhib, FuncS, Apop, In vivo Chimeric (Mouse/Human) Fab
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

Humanized Antibody

CAT Product Name Application Type
TAB-274MZ Human Anti-EGFR Recombinant Antibody (TAB-274MZ) FC Humanized IgG
TAB-297MZ Anti-Human EGFR Recombinant Antibody (H225) ELISA, WB Humanized antibody
TAB-298MZ Anti-Human EGFR Recombinant Antibody (Hu225) ELISA, WB Humanized antibody
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

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-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-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

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-S0023-YC-S(P) Mouse Anti-EGFR Recombinant Antibody (clone 14E1); scFv Fragment IB, FuncS Mouse 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
PSBS-0061 Human Anti-EGFR Recombinant Antibody (Clone FM329); scFv Fragment ELISA, FuncS Human scFv
HPAB-2098-FY-S(P) Mouse Anti-EGFR Recombinant Antibody (clone Mint5); scFv Fragment ICC, Inhib Mouse 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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