Difference between revisions of "Part:BBa K5271000"
 
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<partinfo>BBa_K5271000 short</partinfo>
 
<partinfo>BBa_K5271000 short</partinfo>
  
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The EGFR binding peptide is a 124 amino acid long nanobody with a molecular weight of 13.4 kDa.
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The EGFR-binding peptide is a 124 amino acid long nanobody with a molecular weight of 13.4 kDa that specifically binds to EGFR.
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===Profile===
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*Name: EGFR-binding peptide
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*Base Pairs: 372 bp
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*Amino acid: 124 a.a
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*Origin: ''Llama glama''
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*Properties: EGFR specific nanobody that sterically inhibit the ligands for EGFR activation
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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The epidermal growth factor receptor (EGFR) has been always found to be over-expressed in various type of drug-resistant human cancers, including pancreatic cancer and non-small cell lung cancer. It is a target of antibody-based chemotherapeutics for targeted immunotherapy. This EGFR binding peptide is a 124 amino acid long nanobody with an inhibitory effect on ligand-induced EGFR activation. It inhibits the EGFR activation by a steric blockade on the binding of ligands to EGFR. The EGFR binding peptide consists of the variable domains of heavy chain (VHH) only, which is the smallest antigen-binding modules occurs in the nature. The structure of the nanobody has been resolved using X-ray crystallography [Schmitz ''et al.'', 2013].
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:::https://ars.els-cdn.com/content/image/1-s2.0-S0969212613001664-gr2.jpg
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'''Figure 1. The 7D12 Binding Site on Domain III of sEGFR''' <br>'''(A)''' Cartoon is shown with 7D12 colored green and sEGFRd3 colored gray. CDRs are highlighted in light green and labeled. '''(B)''' In this view, the structure has been rotated ∼180° about a vertical axis relative to (A). The expected locations of domains I, II, and IV of sEGFR are in light blue, based on the structure of tethered sEGFR in PDB ID 1NQL (Ferguson et al., 2003). '''(C)''' View of the interface region between 7D12 and sEGFRd3 in a similar orientation to (A). Side chains that participate in key interactions are shown as sticks, as is the sugar group on sEGFRd3. Predicted salt-bridge (≤4.5 Å) or hydrogen-bond (≤3.5 Å) interactions are indicated with dashed lines. Kabat numbering is used.
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[Excerpt from Schmitz ''et al.'', 2013]
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===Design Note===
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The DNA sequence was obtained from Schmitz ''et al.'', and performed codon optimization for prokaryotic expression system (''E.Coli'' -BL21DE3).
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===Source===
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The VHH domain of the nanobody was isolated from a phage library generated from ''Llama glama'' lymphocytes that had been immunized with A431 epidermoid carcinoma cells [Roovers ''et al.'', 2007]
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The epidermal growth factor receptor (EGFR) has been always found to be over-expressed in various type of drug-resistant human cancers, including pancreatic cancer and non-small cell lung cancer. It is a target of antibody-based chemotherapeutics for targeted immunotherapy. This EGFR binding peptide is a 124 amino acid long nanobody with an inhibitory effect on ligand-induced EGFR activation. It inhibits the EGFR activation by a steric blockade on the binding of ligands to EGFR. The EGFR binding peptide consists of the variable domains of heavy chain (VHH) only, which is the smallest antigen-binding modules occurs in the nature. The structure of the nanobody has been resolved using X-ray crystallography [Schmitz et al., 2013].
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===Reference===
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*Roovers, R. C., Laeremans, T., Huang, L., De Taeye, S., Verkleij, A. J., Revets, H., ... & van Bergen en Henegouwen, P. M. P. (2007). Efficient inhibition of EGFR signalling and of tumour growth by antagonistic anti-EGFR Nanobodies. Cancer immunology, immunotherapy, 56, 303-317.
  
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[[File:https://ars.els-cdn.com/content/image/1-s2.0-S0969212613001664-gr2.jpg|caption]]
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*Schmitz, K. R., Bagchi, A., Roovers, R. C., en Henegouwen, P. M. V. B., & Ferguson, K. M. (2013). Structural evaluation of EGFR inhibition mechanisms for nanobodies/VHH domains. Structure, 21(7), 1214-1224.
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Figure 1. The 7D12 Binding Site on Domain III of sEGFR
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(A) Cartoon is shown with 7D12 colored green and sEGFRd3 colored gray. CDRs are highlighted in light green and labeled. (B) In this view, the structure has been rotated ∼180° about a vertical axis relative to (A). The expected locations of domains I, II, and IV of sEGFR are in light blue, based on the structure of tethered sEGFR in PDB ID 1NQL (Ferguson et al., 2003). (C) View of the interface region between 7D12 and sEGFRd3 in a similar orientation to (A). Side chains that participate in key interactions are shown as sticks, as is the sugar group on sEGFRd3. Predicted salt-bridge (≤4.5 Å) or hydrogen-bond (≤3.5 Å) interactions are indicated with dashed lines. Kabat numbering is used.
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[Excerpt From Schmitz et al., 2013]
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Latest revision as of 09:52, 28 September 2024


EGFR-binding peptide

The EGFR-binding peptide is a 124 amino acid long nanobody with a molecular weight of 13.4 kDa that specifically binds to EGFR.


Profile

  • Name: EGFR-binding peptide
  • Base Pairs: 372 bp
  • Amino acid: 124 a.a
  • Origin: Llama glama
  • Properties: EGFR specific nanobody that sterically inhibit the ligands for EGFR activation


Usage and Biology

The epidermal growth factor receptor (EGFR) has been always found to be over-expressed in various type of drug-resistant human cancers, including pancreatic cancer and non-small cell lung cancer. It is a target of antibody-based chemotherapeutics for targeted immunotherapy. This EGFR binding peptide is a 124 amino acid long nanobody with an inhibitory effect on ligand-induced EGFR activation. It inhibits the EGFR activation by a steric blockade on the binding of ligands to EGFR. The EGFR binding peptide consists of the variable domains of heavy chain (VHH) only, which is the smallest antigen-binding modules occurs in the nature. The structure of the nanobody has been resolved using X-ray crystallography [Schmitz et al., 2013].


1-s2.0-S0969212613001664-gr2.jpg

Figure 1. The 7D12 Binding Site on Domain III of sEGFR
(A) Cartoon is shown with 7D12 colored green and sEGFRd3 colored gray. CDRs are highlighted in light green and labeled. (B) In this view, the structure has been rotated ∼180° about a vertical axis relative to (A). The expected locations of domains I, II, and IV of sEGFR are in light blue, based on the structure of tethered sEGFR in PDB ID 1NQL (Ferguson et al., 2003). (C) View of the interface region between 7D12 and sEGFRd3 in a similar orientation to (A). Side chains that participate in key interactions are shown as sticks, as is the sugar group on sEGFRd3. Predicted salt-bridge (≤4.5 Å) or hydrogen-bond (≤3.5 Å) interactions are indicated with dashed lines. Kabat numbering is used. [Excerpt from Schmitz et al., 2013]


Design Note

The DNA sequence was obtained from Schmitz et al., and performed codon optimization for prokaryotic expression system (E.Coli -BL21DE3).


Source

The VHH domain of the nanobody was isolated from a phage library generated from Llama glama lymphocytes that had been immunized with A431 epidermoid carcinoma cells [Roovers et al., 2007]


Reference

  • Roovers, R. C., Laeremans, T., Huang, L., De Taeye, S., Verkleij, A. J., Revets, H., ... & van Bergen en Henegouwen, P. M. P. (2007). Efficient inhibition of EGFR signalling and of tumour growth by antagonistic anti-EGFR Nanobodies. Cancer immunology, immunotherapy, 56, 303-317.
  • Schmitz, K. R., Bagchi, A., Roovers, R. C., en Henegouwen, P. M. V. B., & Ferguson, K. M. (2013). Structural evaluation of EGFR inhibition mechanisms for nanobodies/VHH domains. Structure, 21(7), 1214-1224.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 43
    Illegal AgeI site found at 175
  • 1000
    COMPATIBLE WITH RFC[1000]