Per citar aquest document: http://ddd.uab.cat/record/145955
Elongation of the C-terminal domain of an anti-amyloid β single-chain variable fragment increases its thermodynamic stability and decreases its aggregation tendency
Rivera Hernández, Geovanny (Universitat Autònoma de Barcelona. Departament de Bioquímica i Biologia Molecular)
Marin Argany, Marta (Universitat Autònoma de Barcelona. Departament de Bioquímica i Biologia Molecular)
Blasco Moreno, B. (Universitat Autònoma de Barcelona. Departament de Bioquímica i Biologia Molecular)
Bonet, Jaume (Universitat Pompeu Fabra. Unitat de Recerca en Informàtica Biomèdica)
Oliva Miguel, Baldomero (Universitat Pompeu Fabra. Unitat de Recerca en Informàtica Biomèdica)
Villegas Hernández, Sandra (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)

Data: 2013
Resum: 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 eningoencephalitis or cerebral hemorrhage. ScFv-h3D6 is known to preclude cytotoxicity of the Aβ 1-42 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.
Nota: This work was supported by Instituto de Salud Carlos III (FIS-PI10–00975), Generalitat de Catalunya (SGR 2009–00761), FEDER (BIO2011–22568) and MICIN (BES-2009–024653). G.R-H is supported by a MAEC-AECI fellowship and M.M-A by a PIF (UAB) fellowship.
Nota: Número d'acord de subvenció ISCIII/FEDER/PI10/00975
Nota: Número d'acord de subvenció AGAUR/2009-SGR-00761
Nota: Número d'acord de subvenció MINECO/BIO2011-22568
Nota: Número d'acord de subvenció MICIN/BES-2009–024653
Drets: Tots els drets reservats.
Llengua: Anglès
Document: article ; recerca ; submittedVersion
Matèria: Alzheimer disease ; CD ; FTIR ; Aggregation ; Immunotherapy ; ScFv ; Stability
Publicat a: mAbs, Vol. 5 Issue 5 (Sep.-Oct. 2013) , p. 678-689, ISSN 1942-0862

DOI: 10.4161/mabs.25382


Pre-print
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