Elongation of the C-terminal domain of an anti-amyloid β single-chain variable fragment increases its thermodynamic stability and decreases its aggregation tendency

Geovanny Rivera-Hernández, Marta Marin-Argany, Bernat Blasco-Moreno, Jaume Bonet, Baldo Oliva, Sandra Villegas

Research output: Contribution to journalArticleResearchpeer-review

15 Citations (Scopus)

Abstract

Amyloid β (Aβ) immunotherapy is considered a promising approach to Alzheimer disease treatment. In contrast to the use of complete antibodies, administration of single-chain variable fragments (scFv) has not been associated with either meningoencephalitis or cerebral hemorrhage. ScFv-h3D6 is known to preclude cytotoxicity of the Aβ1-42 peptide by removing its oligomers from the amyloid pathway. As is the case for other scFv molecules, the recombinant production of scFv-h3D6 is limited by its folding and stability properties. Here, we show that its urea-induced unfolding pathway is characterized by the presence of an intermediate state composed of the unfolded VL domain and the folded VH domain, which suggests the VL domain as a target for thermodynamic stability redesign. The modeling of the 3D structure revealed that the VL domain, located at the C-terminal of the molecule, was ending before its latest β-strand was completed. Three elongation mutants, beyond VL-K107, showed increased thermodynamic stability and lower aggregation tendency, as determined from urea denaturation experiments and Fourier-transform infrared spectroscopy, respectively. Because the mutants maintained the capability of removing Aβ-oligomers from the amyloid pathway, we expect these traits to increase the half-life of scFv-h3D6 in vivo and, consequently, to decrease the effective doses. Our results led to the improvement of a potential Alzheimer disease treatment and may be extrapolated to other class-I scFv molecules of therapeutic interest. © 2013 Landes Bioscience.
Original languageEnglish
Pages (from-to)678-689
JournalmAbs
Volume5
DOIs
Publication statusPublished - 1 Sep 2013

Keywords

  • Aggregation
  • Alzheimer disease
  • CD
  • FTIR
  • Immunotherapy
  • ScFv
  • Stability

Fingerprint Dive into the research topics of 'Elongation of the C-terminal domain of an anti-amyloid β single-chain variable fragment increases its thermodynamic stability and decreases its aggregation tendency'. Together they form a unique fingerprint.

  • Cite this