%0 Journal Article
%T Single-Molecule Two-Color Coincidence Detection of Unlabeled alpha-Synuclein Aggregates.
%A Chappard A
%A Leighton C
%A Saleeb RS
%A Jeacock K
%A Ball SR
%A Morris K
%A Kantelberg O
%A Lee JE
%A Zacco E
%A Pastore A
%A Sunde M
%A Clarke DJ
%A Downey P
%A Kunath T
%A Horrocks MH
%J Angew Chem Weinheim Bergstr Ger
%V 135
%N 15
%D 2023 Apr 3
%M 38516037
暂无%R 10.1002/ange.202216771
%X 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.<BR>Pathological protein aggregates in neurodegenerative disorders are difficult to characterise using current methods. We present a novel single‐molecule detection method to specifically detect and characterise α‐synuclein aggregates at picomolar concentrations. We demonstrate the ability to detect aggregates in biologically relevant samples.