title: The role of trehalose for metastable state and functional form of recombinant interferon beta-1b.
abstract: Physical and mathematical methods such as fluorescence and circular dichroism spectroscopies as well as MCR-ALS chemometric analysis and Wyman linkage theory were applied to assess the effect of trehalose on stability of refolded IFNβ-1b (interferon beta-1b). An intermediate, showing native-like secondary structure is possibly formed during refolding process of IFNβ-1b. Although the refolded protein showed high biological activity, the mathematical data demonstrated a flexible and mid-stable structure, possibly due to its non-glycosylated form. In the presence of 1.5 M of trehalose an increase of 16 °C was observed in the Tm of the refolded protein from the initial of 35 °C. The stability of IFNβ-1b was possibly improved by preferential exclusion of trehalose and formation of a dense hydration shell around the protein surface. Lower amounts of helical structures (16%) were detected in protein solutions without trehalose. In the presence of trehalose the active structure of IFNβ-1b was probably preserved by formation of a metastable structure which subsequently eased post-refolding processes of the protein.
authors.affiliation: Biotechnology Group, Department of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Fazeli
authors.forename: Seyed Abbas
authors.initials: SA
authors.lastname: Shojaosadati
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Fazeli
authors.forename: Khosrow
authors.initials: K
authors.lastname: Khalifeh
authors.forename: Shohreh
authors.initials: S
authors.lastname: Ariaeenejad
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Moosavi-Movahedi
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000068576:Interferon beta-1b; D016899:Interferon-beta / Q000737:chemistry*; D008958:Models, Molecular; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding / Q000187:drug effects; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013816:Thermodynamics; D014199:Trehalose / Q000737:chemistry*
chemical_list: D000998:Antiviral Agents; D011994:Recombinant Proteins; D012996:Solutions; D000068576:Interferon beta-1b; D016899:Interferon-beta; D014199:Trehalose