PASylation technology improves recombinant interferon-β1b solubility, stability, and biological activity

Appl Microbiol Biotechnol. 2017 Mar;101(5):1975-1987. doi: 10.1007/s00253-016-7944-3. Epub 2016 Nov 10.

Abstract

Recombinant interferon-β1b (IFN-β1b) is an effective remedy against multiple sclerosis and other diseases. However, use of small polypeptide (molecular weight is around 18.5 kDa) is limited due to poor solubility, stability, and short half-life in systemic circulation. To solve this problem, we constructed two variants of PASylated IFN-β1b, with PAS sequence at C- or N-terminus of IFN-β1b. The PAS-modified proteins demonstrated 4-fold increase in hydrodynamic volume of the molecule combined with 2-fold increase of in vitro biological activity, as well as advanced stability and solubility of the protein in solution as opposed to unmodified IFN-β1b. Our results demonstrate that PASylation has a positive impact on stability, solubility, and functional activity of IFN-β1b and potentially might improve pharmacokinetic properties of the molecule as a therapeutic agent.

Keywords: Biological activity; Expression; Interferon-β1b; PASylation; Solubility; Stability.

MeSH terms

  • Half-Life
  • Humans
  • Immunologic Factors / genetics
  • Immunologic Factors / metabolism*
  • Immunologic Factors / therapeutic use
  • Interferon beta-1b / genetics*
  • Interferon beta-1b / metabolism*
  • Interferon beta-1b / therapeutic use
  • Multiple Sclerosis / drug therapy
  • Protein Processing, Post-Translational*
  • Protein Stability
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Recombinant Proteins / therapeutic use
  • Solubility

Substances

  • Immunologic Factors
  • Recombinant Proteins
  • Interferon beta-1b