Glycoengineering of interferon-β 1a improves its biophysical and pharmacokinetic properties

PLoS One. 2014 May 23;9(5):e96967. doi: 10.1371/journal.pone.0096967. eCollection 2014.

Abstract

The purpose of this study was to develop a biobetter version of recombinant human interferon-β 1a (rhIFN-β 1a) to improve its biophysical properties, such as aggregation, production and stability, and pharmacokinetic properties without jeopardizing its activity. To achieve this, we introduced additional glycosylation into rhIFN-β 1a via site-directed mutagenesis. Glycoengineering of rhIFN-β 1a resulted in a new molecular entity, termed R27T, which was defined as a rhIFN-β mutein with two N-glycosylation sites at 80th (original site) and at an additional 25th amino acid due to a mutation of Thr for Arg at position 27th of rhIFN-β 1a. Glycoengineering had no effect on rhIFN-β ligand-receptor binding, as no loss of specific activity was observed. R27T showed improved stability and had a reduced propensity for aggregation and an increased half-life. Therefore, hyperglycosylated rhIFN-β could be a biobetter version of rhIFN-β 1a with a potential for use as a drug against multiple sclerosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Biophysical Phenomena*
  • Glycosylation
  • Humans
  • Interferon beta-1a
  • Interferon-beta / chemistry
  • Interferon-beta / genetics*
  • Interferon-beta / metabolism
  • Interferon-beta / pharmacokinetics*
  • Ligands
  • Monosaccharides / metabolism
  • Mutagenesis, Site-Directed*
  • N-Acetylneuraminic Acid / metabolism
  • Protein Conformation
  • Protein Stability
  • Rats
  • Solubility
  • Substrate Specificity

Substances

  • Ligands
  • Monosaccharides
  • Interferon-beta
  • N-Acetylneuraminic Acid
  • Interferon beta-1a

Grants and funding

This research was supported by Basic Science Research Program through the National Research Foundation (NRF)(NRF-2012R1A1A1043288, URL https://www.nrf.re.kr/nrf_tot_cms/index.jsp?pmi-sso-return2=none) and Project through Korea Drug Development Fund (20100030032, URL http://www.kddf.org/Main/) of Korea funded by the Ministry of Education, Science and Technology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.