| Autor: |
Arter WE; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K.; Cavendish Laboratory, University of Cambridge, JJ Thomson Avenue, Cambridge CB3 0HE, U.K., Xu CK; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Castellana-Cruz M; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Herling TW; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Krainer G; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Saar KL; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Kumita JR; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Dobson CM; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K., Knowles TPJ; Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K.; Cavendish Laboratory, University of Cambridge, JJ Thomson Avenue, Cambridge CB3 0HE, U.K. |
| Abstrakt: |
Oligomers comprised of misfolded proteins are implicated as neurotoxins in the pathogenesis of protein misfolding conditions such as Parkinson's and Alzheimer's diseases. Structural, biophysical, and biochemical characterization of these nanoscale protein assemblies is key to understanding their pathology and the design of therapeutic interventions, yet it is challenging due to their heterogeneous, transient nature and low relative abundance in complex mixtures. Here, we demonstrate separation of heterogeneous populations of oligomeric α-synuclein, a protein central to the pathology of Parkinson's disease, in solution using microfluidic free-flow electrophoresis. We characterize nanoscale structural heterogeneity of transient oligomers on a time scale of seconds, at least 2 orders of magnitude faster than conventional techniques. Furthermore, we utilize our platform to analyze oligomer ζ-potential and probe the immunochemistry of wild-type α-synuclein oligomers. Our findings contribute to an improved characterization of α-synuclein oligomers and demonstrate the application of microchip electrophoresis for the free-solution analysis of biological nanoparticle analytes. |
