Part:BBa_K5271013

Panobody -scrambled

A nanobody composed of two binding regions with a scrambled amino acid sequence that acts as a control for the dual targeting nanobody -Panobody.

Profile

• Name: Panobody -scrambled
• Base Pairs: 777 bp
• Amino acid: 258 a.a
• Origin: Synthetic
• Properties: A dual targeting nanobody with a molecular weight of 26.9 kDa that specifically bind to EGFR and HER2 in Gemcitabine resistant pancreatic cancer cells.

Usage and Biology

Figure . The AlphaFold predicted structure of Nanobody scramble. Red: His-tag; Pink: Nanobody scramble; Grey: linker.

Expression

Figure . Protein expression of Panobody -scrambled. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total scrambled Panobody in 16℃ with 0.5mM IPTG induced fraction for 24 hours; lane 3, Soluble scrambled Panobody in 16℃ with 0.5m IPTG induced fraction for 24 hours; lane 3, Insoluble scrambled Panobody in 16℃ with 0.5mM IPTG induced fraction for 24 hours; lane 5, Total scrambled Panobody in 16℃ with uninduced fraction for 24 hours; lane 6, Soluble scrambled Panobody in 16℃ with uninduced fraction for 24 hours; lane 7, Insoluble scrambled Panobody in 16℃ with uninduced fraction for 24 hours.

Figure . Protein expression of scrambled Panobody. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total scrambled Panobody in 25℃ with 0.5mM IPTG induced fraction for 24 hours; lane 3, Soluble scrambled Panobody in 25℃ with 0.5m IPTG induced fraction for 24 hours; lane 3, Insoluble scrambled Panobody in 25℃ with 0.5mM IPTG induced fraction for 24 hours; lane 5, Total scrambled Panobody in 25℃ with uninduced fraction for 24 hours; lane 6, Soluble scrambled Panobody in 25℃ with uninduced fraction for 24 hours; lane 7, Insoluble scrambled Panobody in 25℃ with uninduced fraction for 24 hours.​

Figure . Protein expression of scrambled Panobody. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total scrambled Panobody in 30℃ with 0.5mM IPTG induced fraction for 24 hours; lane 3, Soluble scrambled Panobody in 30℃ with 0.5m IPTG induced fraction for 24 hours; lane 3, Insoluble scrambled Panobody in 30℃ with 0.5mM IPTG induced fraction for 24 hours; lane 5, Total scrambled Panobody in 30℃ with uninduced fraction for 24 hours; lane 6, Soluble scrambled Panobody in 30℃ with uninduced fraction for 24 hours; lane 7, Insoluble scrambled Panobody in 30℃ with uninduced fraction for 24 hours.​

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.
• 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, Atarhouch T, Muyldermans S. Naturally occurring antibodies devoid of light chains. Nature 1993;363:446–48.
• Vaneycken I, Devoogdt N, Van Gassen N, Vincke C, Xavier C, Wernery U, et al Preclinical screening of anti-HER2 nanobodies for molecular imaging of breast cancer. FASEB J 2011;25:2433–2446.
• Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., ... & Hassabis, D. (2021). Highly accurate protein structure prediction with AlphaFold. nature, 596(7873), 583-589.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 834
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]