Purification of recombinant Aβ(1-42) and pGlu-Aβ(3-42) using preparative SDS-PAGE

Electrophoresis
Claudia SpahnStephan Schilling

Abstract

Recombinant expression and purification of amyloid peptides represents a common basis for investigating the molecular mechanisms of amyloid formation and toxicity. However, the isolation of the recombinant peptides is hampered by inefficient separation from contaminants such as the fusion protein required for efficient expression in E. coli. Here, we present a new approach for the isolation of highly purified Aβ(1-42) and pGlu-Aβ(3-42), which is based on a separation using preparative SDS-PAGE. The method relies on the purification of the Aβ fusion protein by affinity chromatography followed by preparative SDS-PAGE under reducing conditions and subsequent removal of detergents by precipitation. The application of preparative SDS-PAGE represents the key step to isolate highly pure recombinant Aβ, which has been applied for characterization of aggregation and toxicity. Thereby, the yield of the purification strategy was  >60%. To the best of our knowledge, this is the first description of an electrophoresis-based method for purification of a recombinant Aβ peptide. Therefore, the method might be of interest for isolation of other amyloid peptides, which are critical for conventional purification strategies due to their aggregatio...Continue Reading

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Citations

Mar 24, 2020·Critical Reviews in Biotechnology·Longgang JiaFufeng Liu
Feb 6, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Janett KöppenStephan Schilling

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