The aggregation of cytochrome C may be linked to its flexibility during refolding

James Austerberry, Daniel Belton

Research output: Contribution to journalArticlepeer-review


Large-scale expression of biopharmaceutical proteins in cellular hosts results in production of large insoluble mass aggregates. In order to generate functional product, these aggregates require further processing through refolding with denaturant, a process in itself that can result in aggregation. Using a model folding protein, cytochrome C, we show how an increase in final denaturant concentration decreases the propensity of the protein to aggregate during refolding. Using polarised fluorescence anisotropy, we show how reduced levels of aggregation can be achieved by increasing the period of time the protein remains flexible during refolding, mediated through dilution ratios. This highlights the relationship between the flexibility of a protein and its propensity to aggregate. We attribute this behaviour to the preferential urea-residue interaction, over self-association between molecules.
Original languageEnglish
Number of pages4
Journal3 Biotech
Issue number33
Early online date14 Jan 2016
Publication statusPublished - Jun 2016


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