Part:BBa_K4634012

OmpA-PD-L1 nanobody

The OmpA signalling sequence(BBa_K4380001) is a short amino acid sequence found at the N-terminus of the OmpA protein in Gram-negative bacteria. The signal peptide acts as a recognition signal for the protein translocation machinery present in the bacterial cell. When fused to the N-terminus of other proteins, it has the ability to facilitate the secretion of those proteins.

PD-L1 nanobody is a camelid single-domain antibody fragment blocking PD-L1 (programmed cell death–ligand 1). It acts as an immune check point inhibitor, which have revolutionized the paradigm of cancer immunotherapy treatments.

In the composite part OmpA-PD-L1 nanobody, 21aa long OmpA signal peptide is linked at the N terminal of PD-L1 nanobody, which facilitate the secretion of nanobody without changing its structure (predicted by Alphafold).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 316
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 316
  Illegal NotI site found at 269
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 316
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 316
  Illegal AgeI site found at 94
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Gurbatri CR, Lia I, Vincent R, Coker C, Castro S, Treuting PM, Hinchliffe TE, Arpaia N, Danino T. Engineered probiotics for local tumor delivery of checkpoint blockade nanobodies. Sci Transl Med. 2020 Feb 12;12(530):eaax0876. doi: 10.1126/scitranslmed.aax0876. PMID: 32051224; PMCID: PMC7685004.
[2] Movva NR, Nakamura K, Inouye M. Amino acid sequence of the signal peptide of ompA protein, a major outer membrane protein of Escherichia coli. J Biol Chem. 1980 Jan 10;255(1):27-9. PMID: 6985608.