title: Separation and detection of individual Aβ aggregates by capillary electrophoresis with laser-induced fluorescence detection.
abstract: The separation and detection of individual amyloid beta (Aβ) aggregates by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was demonstrated. Samples were prepared with either Aβ (1-40) or Aβ (1-42) peptides and were characterized by CE with ultraviolet (UV) absorbance detection and transmission electron microscopy (TEM). Using thioflavin T (ThT) in the electrophoresis buffer, electrophoresis of aggregate-containing samples (5.0-s injection) produced up to several hundred narrow (< 20 ms FWHM [full width at half maximum]) fluorescence peaks. Injection of Aβ (1-40) monomer samples resulted in no additional peaks compared with controls. The CE-LIF results were validated by bulk ThT fluorescence measurements for the same samples. The potential of laser-induced fluorescence anisotropy (LIFA) with CE to characterize individual Aβ aggregates also was investigated.
authors.affiliation: Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803, USA.
authors.forename: Ryan A
authors.initials: RA
authors.lastname: Picou
authors.forename: David P
authors.initials: DP
authors.lastname: Schrum
authors.forename: Ginger
authors.initials: G
authors.lastname: Ku
authors.forename: Rebekah A
authors.initials: RA
authors.lastname: Cerqua
authors.forename: Indu
authors.initials: I
authors.lastname: Kheterpal
authors.forename: S Douglass
authors.initials: SD
authors.lastname: Gilman
mesh_terms: D016229:Amyloid beta-Peptides / Q000032:analysis* / Q000302:isolation & purification; D052160:Benzothiazoles; D019075:Electrophoresis, Capillary; D007834:Lasers; D017348:Microscopy, Electron, Scanning Transmission; D010446:Peptide Fragments / Q000032:analysis* / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet; D013844:Thiazoles / Q000737:chemistry
chemical_list: D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; D010446:Peptide Fragments; D013844:Thiazoles; C078592:amyloid beta-protein (1-40); C075222:amyloid beta-protein (1-42); C009462:thioflavin T