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Rapid Fractionation and Characterisation of Alpha-Synuclein Oligomers in Solution

Arter, W. E.; Xu, C. K.; Castellana-Cruz, M.; Herling, T. W.; Krainer, G.; Saar, K. L.; Kumita, J. R.; Dobson, C. M.; Knowles, T. P.

2020-03-14 biophysics
10.1101/2020.03.10.985804 bioRxiv
Show abstract

Oligomeric intermediates are implicated as neurotoxins in the pathogenesis of protein mis-folding diseases. Structural, biophysical and biochemical characterisation of these species is challenging due to their heterogeneous and transient nature, and their typically low abun-dance. Here, we show that microfluidic free-flow electrophoresis is capable of separating heterogeneous oligomer mixtures on a timescale of seconds, at least two orders of magnitude faster than conventional techniques. This enables analysis of oligomer structural heterogeneity, zeta-potential and immunochemistry with minimal sample perturbation under physiologically-relevant conditions.

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