Structural differences of amyloid-β fibrils revealed by antibodies from phage display

Patrick Droste, André Frenzel, Miriam Steinwand, Thibaut Pelat, Philippe Thullier, Michael Hust, Hilal Lashuel, Stefan Dübel

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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 aα-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.

Original languageEnglish
Article number57
JournalBMC Biotechnology
Volume15
Issue number1
DOIs
Publication statusPublished - 18 Jun 2015
Externally publishedYes

Fingerprint

Amyloid
Bacteriophages
Antibodies
Libraries
Peptides
Neurofibrillary Tangles
Aptitude
Antibody Affinity
Macaca fascicularis
Amyloid Plaques
Macaca
Gel Chromatography
Epitopes
Immunization
Alzheimer Disease
Hand
Antigens
Brain

Keywords

  • Abeta
  • Alzheimer's disease
  • Beta-amyloid
  • Immune library
  • Phage display
  • scFv

ASJC Scopus subject areas

  • Biotechnology

Cite this

Droste, P., Frenzel, A., Steinwand, M., Pelat, T., Thullier, P., Hust, M., ... Dübel, S. (2015). Structural differences of amyloid-β fibrils revealed by antibodies from phage display. BMC Biotechnology, 15(1), [57]. https://doi.org/10.1186/s12896-015-0146-8

Structural differences of amyloid-β fibrils revealed by antibodies from phage display. / Droste, Patrick; Frenzel, André; Steinwand, Miriam; Pelat, Thibaut; Thullier, Philippe; Hust, Michael; Lashuel, Hilal; Dübel, Stefan.

In: BMC Biotechnology, Vol. 15, No. 1, 57, 18.06.2015.

Research output: Contribution to journalArticle

Droste, P, Frenzel, A, Steinwand, M, Pelat, T, Thullier, P, Hust, M, Lashuel, H & Dübel, S 2015, 'Structural differences of amyloid-β fibrils revealed by antibodies from phage display', BMC Biotechnology, vol. 15, no. 1, 57. https://doi.org/10.1186/s12896-015-0146-8
Droste P, Frenzel A, Steinwand M, Pelat T, Thullier P, Hust M et al. Structural differences of amyloid-β fibrils revealed by antibodies from phage display. BMC Biotechnology. 2015 Jun 18;15(1). 57. https://doi.org/10.1186/s12896-015-0146-8
Droste, Patrick ; Frenzel, André ; Steinwand, Miriam ; Pelat, Thibaut ; Thullier, Philippe ; Hust, Michael ; Lashuel, Hilal ; Dübel, Stefan. / Structural differences of amyloid-β fibrils revealed by antibodies from phage display. In: BMC Biotechnology. 2015 ; Vol. 15, No. 1.
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