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
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Binding Sites
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Biophysical Phenomena*
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Glycosylation
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Humans
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Interferon beta-1a
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Interferon-beta / chemistry
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Interferon-beta / genetics*
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Interferon-beta / metabolism
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Interferon-beta / pharmacokinetics*
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Ligands
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Monosaccharides / metabolism
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Mutagenesis, Site-Directed*
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N-Acetylneuraminic Acid / metabolism
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Protein Conformation
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Protein Stability
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Rats
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Solubility
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Substrate Specificity
Substances
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Ligands
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Monosaccharides
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Interferon-beta
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N-Acetylneuraminic Acid
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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.