Part:BBa_K5271001

HER2-binding peptide

The HER2 binding peptide is a 115 amino acid long nanobody with a molecular weight of 12.6 kDa that specifically bind to HER2.

Profile

• Name: HER2-binding peptide
• Base Pairs: 345 bp
• Amino acid: 115 a.a
• Origin: Camelus dromedarius
• Properties: A HER2 specific nanobody that binds to domain I of HER2 with a non-competitive nature to mAB drugs, trastuzumab and pertuzumab for HER2 targeting.

Usage and Biology

The human epidermal growth factor receptor 2 (HER2), is a transmembrane glycoprotein receptor that has an important role in cell proliferation and survival. Over-expression of HER2 associates with aggressive, metastatic and drug-resistant tumor phenotype. Immunotherapy targeting HER2 using monoclonal antibody (mAb) can significantly increase the survival rate of patients, yet, resistance to mAB-based drugs is frequently developed in HER+ cancers. Nanobody is an antigen binding fragments that are derived from the heavy-chain of antibodies. HER2-binding peptide is derived from Camelus dromedarius and have advantages of small size, higher stability and exceptional binding specificity than conventional mAb. [D'Huyvetter et al., 2017] The HER2 nanobody binds to an epitope on HER2 domain I, which is distinct from the HER2 recognition sites of conventional mAb such as pertuzumab and trastuzumab (Fig. 1). The interactions with HER2 are mediated by the complementarity determining regions (CDR) and the framework regions (FR). The binding of this nanobody has non-competitive nature to mAB drug, trastuzumab and pertuzumab for HER2 targeting. [Vaneycken et al., 2011] The structure of the nanobody has been resolved using X-ray crystallography. [D'Huyvetter et al., 2017]

Figure 1. Structure of 2Rs15d (cartoon representation) complexed with HER2(1-646)His (surface representation).
(A) 2Rs15d (red) binds HER2 domain I (tan; Gln2-Arg196), while pertuzumab and trastuzumab interact with domain II (sky blue; Thr197–Val320) and domain IV (sandy brown; Cys490–Cys566), respectively. HER2 domain III (Cys321–Ala489) is colored in plum. [Excerpt from D'Huyvetter et al., 2017]

Design Note

The DNA sequence was obtained from D'Huyvetter et al., and performed condon optimization for prokaryotic expression system (E.Coli -BL21DE3).

Source

The nanobody is an antigen-binding fragments that are derived from Camelus dromedarius heavy-chain antibodies and have advantageous characteristics compared with mAbs and their derived fragments. [Hamers-Casterman et al., 1993]

Reference

• D'Huyvetter, M., De Vos, J., Xavier, C., Pruszynski, M., Sterckx, Y. G., Massa, S., ... & Devoogdt, N. (2017). 131I-labeled anti-HER2 camelid sdAb as a theranostic tool in cancer treatment. Clinical cancer research, 23(21), 6616-6628.

• Hamers-Casterman, C. T. S. G., Atarhouch, T., Muyldermans, S. A., Robinson, G., Hammers, C., Songa, E. B., ... & Hammers, R. (1993). Naturally occurring antibodies devoid of light chains. Nature, 363(6428), 446-448.

• Vaneycken, I., Devoogdt, N., Van Gassen, N., Vincke, C., Xavier, C., Wernery, U., ... & Caveliers, V. (2011). Preclinical screening of anti‐HER2 nanobodies for molecular imaging of breast cancer. The FASEB Journal, 25(7), 2433-2446.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 73
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]