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Production of Recombinant Alpha-Synuclein: Still No Standardized Protocol in Sight

Lookup NU author(s): Professor Tiago OuteiroORCiD



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


© 2022 by the authors. Licensee MDPI, Basel, Switzerland. Synucleinopathies are a group of neurodegenerative diseases, characterized by the abnormal accumulation of the protein alpha-synuclein (aSyn). aSyn is an intrinsically disordered protein that can adopt different aggregation states, some of which may be associated with disease. Therefore, understanding the transitions between such aggregation states may be essential for deciphering the molecular underpinnings underlying synucleinopathies. Recombinant aSyn is routinely produced and purified from E. coli in many laboratories, and in vitro preparations of aSyn aggregated species became central for modeling neurodegeneration in cell and animal models. Thus, reproducibility and reliability of such studies largely depends on the purity and homogeneity of aSyn preparations across batches and between laboratories. A variety of different methods are in use to produce and purify aSyn, which we review in this commentary. We also show how extraction buffer composition can affect aSyn aggregation, emphasizing the importance of standardizing protocols to ensure reproducibility between different laboratories and studies, which are essential for advancing the field.

Publication metadata

Author(s): Al-Azzani M, Konig A, Outeiro TF

Publication type: Article

Publication status: Published

Journal: Biomolecules

Year: 2022

Volume: 12

Issue: 2

Online publication date: 18/02/2022

Acceptance date: 16/02/2022

Date deposited: 10/03/2022

ISSN (electronic): 2218-273X

Publisher: MDPI


DOI: 10.3390/biom12020324

PubMed id: 35204823


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Funder referenceFunder name
EXC 2067/1-390729940
SFB1286 Project B8