title: Analysis of Aβ (1-40) and Aβ (1-42) monomer and fibrils by capillary electrophoresis.
abstract: A method based on capillary electrophoresis (CE) with UV absorbance detection is presented to characterize synthetic amyloid beta (Aβ) peptide preparations at different aggregation states. Aggregation of Aβ (1-40) and Aβ (1-42) is closely linked to Alzheimer's disease (AD), and studying how Aβ peptides self-assemble to form aggregates is the focus of intense research. Developing methods capable of identifying, characterizing and quantifying a wide range of Aβ species from monomers to fully formed fibrils is critical for AD research and is a major analytical challenge. Monomer and fibril samples of Aβ (1-40) and Aβ (1-42) were prepared and characterized for this study. The monomer-equivalent concentration for each sample was determined by HPLC-UV, and aggregate formation was confirmed and characterized by transmission electron microscopy. The same samples were studied using CE with UV absorbance detection. Analysis by mass spectrometry of collected CE fractions was used to confirm the presence of Aβ for some CE-UV peaks. The CE-UV method reported here clearly indicates that monomeric and aggregated Aβ were electrophoretically separated, and substantial differences in the electrophoretic profiles between samples of Aβ (1-40) and Aβ (1-42) were observed. This CE-UV method can differentiate between Aβ monomer, oligomeric intermediates, and mature fibrils.
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: Indu
authors.initials: I
authors.lastname: Kheterpal
authors.forename: Amber D
authors.initials: AD
authors.lastname: Wellman
authors.forename: Madhavi
authors.initials: M
authors.lastname: Minnamreddy
authors.forename: Ginger
authors.initials: G
authors.lastname: Ku
authors.forename: S Douglass
authors.initials: SD
authors.lastname: Gilman
mesh_terms: D016229:Amyloid beta-Peptides / Q000032:analysis* / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D055503:Protein Multimerization; D021122:Protein Subunits; D013056:Spectrophotometry, Ultraviolet
chemical_list: D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; D021122:Protein Subunits; C078592:amyloid beta-protein (1-40); C075222:amyloid beta-protein (1-42)