Modular Conjugation of a Potent Anti-HER2 Immunotoxin Using Coassociating Peptides - Archive ouverte HAL Access content directly
Journal Articles Bioconjugate Chemistry Year : 2020

Modular Conjugation of a Potent Anti-HER2 Immunotoxin Using Coassociating Peptides

Audrey Stoessel
  • Function : Author
Nadja Groysbeck
  • Function : Author
  • PersonId : 1081553
Lucile Guyot
  • Function : Author
Lina Barret
  • Function : Author
Ambre Bender
  • Function : Author
  • PersonId : 1074170
Thomas Lutz
  • Function : Author
Nikita Pallaoro
  • Function : Author
Marie Blocat
  • Function : Author
Guy Zuber
  • Function : Author
  • PersonId : 1065490
Bruno Chatton
  • Function : Author
  • PersonId : 1076869
Mariel Donzeau
  • Function : Author
  • PersonId : 1080228

Abstract

Immunotoxins are emerging candidates for cancer therapeutics. These biomolecules consist of a cell targeting protein combined to a polypeptide toxin. Associations of both entities can be achieved either chemically by covalent bonds or genetically creating fusion proteins. However, chemical agents can affect activity and/or stability of the conjugate proteins and additional purification steps are often required to isolate the final conjugate from unwanted by-products. As for fusion proteins, they often suffer from low solubility and yield. In this report, we describe a straightforward conjugation process to generate an immunotoxin using co-associating peptides (named K3 and E3), originating from the tetramerization domain of p53. To that end, a nanobody targeting the human epidermal growth factor receptor 2 (nano-HER2) and a protein toxin fragment from Pseudomonas aeruginosa Exotoxin A (TOX) were genetically fused to the E3 and K3 peptides. Entities were produced separately in E. coli in soluble forms and at high yields. The nano-HER2 fused to the E3 or K3 helixes (nano-HER2-E3 and nano-HER2-K3) and the co-assembled immunotoxins (nano-HER2-K3E3-TOX and nano-HER2-E3K3-TOX) presented binding specificity on HER2 overexpressing cells with relative binding constants in the low nanomolar to picomolar range. Both toxin modules (E3-TOX and K3-TOX) and the combined immunotoxins exhibited similar cytotoxicity levels compared to the toxin alone (TOX). Finally, nano-HER2-K3E3-TOX and nano-HER2-E3K3-TOX evaluated on various breast cancer cells were highly potent and specific to kill HER2-overexpressing breast cancer cells with IC50 values in the picomolar range. Altogether, we demonstrate that this non-covalent conjugation method using two co-assembling peptides can be easily implemented for modular engineering of immunotoxins targeting different types of cancers.

Domains

Bioengineering
Fichier principal
Vignette du fichier
islandora_105415.pdf (2.93 Mo) Télécharger le fichier
Origin : Files produced by the author(s)

Dates and versions

hal-03005245 , version 1 (13-11-2020)

Identifiers

Cite

Audrey Stoessel, Nadja Groysbeck, Lucile Guyot, Lina Barret, Yves Nominé, et al.. Modular Conjugation of a Potent Anti-HER2 Immunotoxin Using Coassociating Peptides. Bioconjugate Chemistry, 2020, 31 (10), pp.2421-2430. ⟨10.1021/acs.bioconjchem.0c00482⟩. ⟨hal-03005245⟩
49 View
142 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More