Korean Journal of Chemical Engineering

, Volume 29, Issue 6, pp 693–702

Techniques for monitoring protein misfolding and aggregation in vitro and in living cells

Authors

  • Simpson Gregoire
    • Department of Chemical EngineeringUniversity of Virginia
  • Jacob Irwin
    • Department of Chemical EngineeringUniversity of Virginia
    • Department of Chemical EngineeringUniversity of Virginia
    • Institutes on AgingUniversity of Virginia
Invited Review Paper

DOI: 10.1007/s11814-012-0060-x

Cite this article as:
Gregoire, S., Irwin, J. & Kwon, I. Korean J. Chem. Eng. (2012) 29: 693. doi:10.1007/s11814-012-0060-x

Abstract

Protein misfolding and aggregation have been considered important in understanding many neurodegenerative diseases and recombinant biopharmaceutical production. Various traditional and modern techniques have been utilized to monitor protein aggregation in vitro and in living cells. Fibril formation, morphology and secondary structure content of amyloidogenic proteins in vitro have been monitored by molecular probes, TEM/AFM, and CD/FTIR analyses, respectively. Protein aggregation in living cells has been qualitatively or quantitatively monitored by numerous molecular folding reporters based on either fluorescent protein or enzyme. Aggregation of a target protein is directly correlated to the changes in fluorescence or enzyme activity of the folding reporter fused to the target protein, which allows non-invasive monitoring aggregation of the target protein in living cells. Advances in the techniques used to monitor protein aggregation in vitro and in living cells have greatly facilitated the understanding of the molecular mechanism of amyloidogenic protein aggregation associated with neurodegenerative diseases, optimizing culture conditions to reduce aggregation of biopharmaceuticals expressed in living cells, and screening of small molecule libraries in the search for protein aggregation inhibitors.

Key words

Protein MisfoldingAggregationNeurodegenerative DiseasesAmyloid Fibrils

Copyright information

© Korean Institute of Chemical Engineers, Seoul, Korea 2012