Authors:

Samhita Mysore Shantharam

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About Paper:

Parkinson's Disease is a devastating neurodegenerative disease characterized by lesions in the brain known as Lewy bodies. These lesions are the result of aggregation of the alpha-synuclein protein. The mechanisms that lead to these harmful lesions are still unclear and the vast majority of research has focused on identifying the cause. This lack of clarity poses incredible difficulty with diagnosis of PD, with confirmation only occurring upon post-mortem analysis. Seed Amplification Assay (SAA) is a relatively new diagnostic method that replicates the aggregation in vitro through a cyclical process and can be used with cerebral spinal fluid samples from patients. The addition of a highly concentrated solution of recombinant alpha-synuclein will catalyze the aggregation within a real-time quake-inducing conversion (RT-QuIC) system but a negative control is needed in order to reduce inconsistencies and noise with the data output. The current work of the project consists of purifying the K23Q strain of alpha-synuclein to act as a negative control for this experiment. The K23Q strain has been identified recently to be a relatively stagnant strain that resists aggregation. Protein purification consists of outgrowth in E. coli bacteria, protein lysis, size exclusion chromatography, and anionic exchange chromatography. Together, these techniques were implemented to extract and purify K23Q and improve diagnostic accuracy of SAA. Keywords: [no keywords provided]

Source:

Purdue University / 2025

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Co-authors:

Samhita Mysore Shantharam