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- Title
Single‐Molecule Two‐Color Coincidence Detection of Unlabeled alpha‐Synuclein Aggregates.
- Authors
Chappard, Alexandre; Leighton, Craig; Saleeb, Rebecca S.; Jeacock, Kiani; Ball, Sarah R.; Morris, Katie; Kantelberg, Owen; Lee, Ji‐Eun; Zacco, Elsa; Pastore, Annalisa; Sunde, Margaret; Clarke, David J.; Downey, Patrick; Kunath, Tilo; Horrocks, Mathew H.
- Abstract
Protein misfolding and aggregation into oligomeric and fibrillar structures is a common feature of many neurogenerative disorders. Single‐molecule techniques have enabled characterization of these lowly abundant, highly heterogeneous protein aggregates, previously inaccessible using ensemble averaging techniques. However, they usually rely on the use of recombinantly‐expressed labeled protein, or on the addition of amyloid stains that are not protein‐specific. To circumvent these challenges, we have made use of a high affinity antibody labeled with orthogonal fluorophores combined with fast‐flow microfluidics and single‐molecule confocal microscopy to specifically detect α‐synuclein, the protein associated with Parkinson's disease. We used this approach to determine the number and size of α‐synuclein aggregates down to picomolar concentrations in biologically relevant samples.
- Subjects
COINCIDENCE circuits; ALPHA-synuclein; PARKINSON'S disease; CONFOCAL microscopy
- Publication
Angewandte Chemie, 2023, Vol 135, Issue 15, p1
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.202216771