A screening methodology for purifying proteins with aggregation problems

Mario Lebendiker*, Michal Maes, Assaf Friedler

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Many proteins are prone to aggregate or insoluble for different reasons. This poses an extraordinary challenge at the expression level, but even more during downstream purification processes. Here we describe a strategy that we developed for purifying prone-to-aggregate proteins. Our methodology can be easily implemented in small laboratories without the need for automated, expensive platforms. This procedure is especially suitable for intrinsically disordered proteins (IDPs) and for proteins with intrinsically disordered regions (IDRs). Such proteins are likely to aggregate due to their lack of tertiary structure and their extended and fl exible conformations. Similar methodologies can be applied to other proteins with comparable tendency to aggregate during the expression or purification steps. In this chapter, we will mainly focus on protein solubility and stability issues during purification and storage, on factors that can prevent aggregation or maintain solubility, and on the importance of the early elimination of aggregates during protein purification.

Original languageAmerican English
Pages (from-to)261-281
Number of pages21
JournalMethods in Molecular Biology
StatePublished - 2015

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media New York 2015.


  • Aggregation analysis
  • Aggregation suppressors
  • Buffer conditions
  • Chaotropes
  • Insoluble proteins
  • Intrinsically disordered proteins
  • Kosmotropes
  • Protein aggregation
  • Protein concentration
  • Protein storage
  • Stabilizers


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