KCS

α-Synuclein aggregates are the pathological hallmarks of synucleinopathies, including Parkinson’s disease and dementia with Lewy bodies. However, there is still no treatment for the synucleinopathies that has been established. To develop a therapeutic strategy, it is crucial to understand the molecular basis of α-synuclein aggregation and suppress the pathogenic amyloid aggregate formation. Up to date, as in-human studies are limited, the majority of research findings were based on in vitro experiments, with purified recombinant α-synuclein used to investigate self-assembly properties. Herein, we developed high-purity recombinant full-length α-synuclein expression and purification protocols without affinity tags or linkers. Furthermore, we performed a quality assessment of purified recombinant proteins to confirm the unique properties of α-synuclein via biophysical analyses including mass spectrometry-based identification, thioflavin T (ThT) fluorescence assay, and circular dichroism (CD). The compelling results imply that the proposed non-tagged α-synuclein preserves its biophysical properties, which can be used in further study to investigate the self-assembly property of α-synuclein.