Structural differences of amyloid-β fibrils revealed by antibodies from phage display.
- Droste P Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics, Spielmannstr.7, 38106, Braunschweig, Germany. Patrick.droste@celerion.com.
- Frenzel A Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics, Spielmannstr.7, 38106, Braunschweig, Germany. andre.frenzel@tu-bs.de.
- Steinwand M Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics, Spielmannstr.7, 38106, Braunschweig, Germany. Miriam.steinwand@delenex.com.
- Pelat T Institut de recherche Biomédicale des Armées (IRBA-CRSSA); Département de Microbiologie; Unité de biotechnologie des anticorps et des toxines, La Tronche Cedex, France. thibaut.pelat@biotem.fr.
- Thullier P Institut de recherche Biomédicale des Armées (IRBA-CRSSA); Département de Microbiologie; Unité de biotechnologie des anticorps et des toxines, La Tronche Cedex, France. pthullier@yahoo.com.
- Hust M Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics, Spielmannstr.7, 38106, Braunschweig, Germany. m.hust@tu-bs.de.
- Lashuel H SV-BMI, Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute, École Polytechnique Fédérale de Lausanne, Station 19, 1015, Lausanne, Switzerland. hilal.lashuel@epfl.ch.
- Dübel S Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics, Spielmannstr.7, 38106, Braunschweig, Germany. s.duebel@tu-bs.de.
- 2015-06-19
Published in:
- BMC biotechnology. - 2015
Alzheimer Disease
Amino Acid Sequence
Amyloid
Amyloid beta-Peptides
Antibodies
Epitopes
Humans
Peptide Fragments
Protein Aggregation, Pathological
Protein Conformation
Protein Structure, Secondary
English
BACKGROUND
Beside neurofibrillary tangles, amyloid plaques are the major histological hallmarks of Alzheimer's disease (AD) being composed of aggregated fibrils of β-amyloid (Aβ). During the underlying fibrillogenic pathway, starting from a surplus of soluble Aβ and leading to mature fibrils, multiple conformations of this peptide appear, including oligomers of various shapes and sizes. To further investigate the fibrillization of β-amyloid and to have tools at hand to monitor the distribution of aggregates in the brain or even act as disease modulators, it is essential to develop highly sensitive antibodies that can discriminate between diverse aggregates of Aβ.
RESULTS
Here we report the generation and characterization of a variety of amyloid-β specific human and human-like antibodies. Distinct fractions of monomers and oligomers of various sizes were separated by size exclusion chromatography (SEC) from Aβ42 peptides. These antigens were used for the generation of two Aβ42 specific immune scFv phage display libraries from macaque (Macaca fascicularis). Screening of these libraries as well as two naïve human phage display libraries resulted in multiple unique binders specific for amyloid-β. Three of the obtained antibodies target the N-terminal part of Aβ42 although with varying epitopes, while another scFv binds to the α-helical central region of the peptide. The affinities of the antibodies to various Aβ42 aggregates as well as their ability to interfere with fibril formation and disaggregation of preformed fibrils were determined. Most significantly, one of the scFv is fibril-specific and can discriminate between two different fibril forms resulting from variations in the acidity of the milieu during fibrillogenesis.
CONCLUSION
We demonstrated that the approach of animal immunization and subsequent phage display based antibody selection is applicable to generate highly specific anti β-amyloid scFvs that are capable of accurately discriminating between minute conformational differences.
Beside neurofibrillary tangles, amyloid plaques are the major histological hallmarks of Alzheimer's disease (AD) being composed of aggregated fibrils of β-amyloid (Aβ). During the underlying fibrillogenic pathway, starting from a surplus of soluble Aβ and leading to mature fibrils, multiple conformations of this peptide appear, including oligomers of various shapes and sizes. To further investigate the fibrillization of β-amyloid and to have tools at hand to monitor the distribution of aggregates in the brain or even act as disease modulators, it is essential to develop highly sensitive antibodies that can discriminate between diverse aggregates of Aβ.
RESULTS
Here we report the generation and characterization of a variety of amyloid-β specific human and human-like antibodies. Distinct fractions of monomers and oligomers of various sizes were separated by size exclusion chromatography (SEC) from Aβ42 peptides. These antigens were used for the generation of two Aβ42 specific immune scFv phage display libraries from macaque (Macaca fascicularis). Screening of these libraries as well as two naïve human phage display libraries resulted in multiple unique binders specific for amyloid-β. Three of the obtained antibodies target the N-terminal part of Aβ42 although with varying epitopes, while another scFv binds to the α-helical central region of the peptide. The affinities of the antibodies to various Aβ42 aggregates as well as their ability to interfere with fibril formation and disaggregation of preformed fibrils were determined. Most significantly, one of the scFv is fibril-specific and can discriminate between two different fibril forms resulting from variations in the acidity of the milieu during fibrillogenesis.
CONCLUSION
We demonstrated that the approach of animal immunization and subsequent phage display based antibody selection is applicable to generate highly specific anti β-amyloid scFvs that are capable of accurately discriminating between minute conformational differences.
- Language
-
- English
- Open access status
- gold
- Identifiers
-
- DOI 10.1186/s12896-015-0146-8
- PMID 26084577
- Persistent URL
- https://sonar.ch/global/documents/69688
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