Structural analysis of modified forms of recombinant IFN-β produced under stress-simulating conditions

Stefania Orrù, Angela Amoresano, Rosa Siciliano, Roberta Napoleoni, Ornella Finocchiaro, Antonio Datola, Eva De Luca, Antonino Sirna, Piero Pucci

Research output: Contribution to journalArticlepeer-review


The present study focused on the investigation of the chemical stability of recombinant human interferon-β (rhIFN-β) tested in vitro by chemical treatments that simulate stress-induced conditions that may occur during handling, storage or ageing of protein samples. Mild oxidation and/or alkylation of the recombinant protein showed that the four methionines occurring in the interferon displayed different chemical susceptibility in that Met36 and Met117 were fully modified, whereas Met1 showed only little modification and Met62 was completely resistant. Moreover, incubation of rhIFN-β under alkaline conditions resulted in the formation of a covalent dimeric species stabilised by an intermolecular disulphide bridge involving the free SH group of Cys17 from each polypeptide chain. Analysis of biological activity of the different IFN-β derivatives showed that rhIFN-β fully retains its specific activity following mild oxidation treatments whereas reaction with a high concentration of alkylating agents or incubation under alkaline conditions strongly reduce its specific antiviral activity.

Original languageEnglish
Pages (from-to)7-17
Number of pages11
JournalBiological Chemistry
Issue number1
Publication statusPublished - Jan 2000


  • β-interferon
  • Glycoproteins
  • Mass spectrometry
  • Stress-induced modifications
  • Structural analysis

ASJC Scopus subject areas

  • Biochemistry


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