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

• Published in: PloS one Vol. 9; no. 5; p. e96967
• Main Authors: Song, Kyoung, Yoon, In-Soo, Kim, Nam Ah, Kim, Dong-Hwan, Lee, Jongmin, Lee, Hee Jung, Lee, Saehyung, Choi, Sunghyun, Choi, Min-Koo, Kim, Ha Hyung, Jeong, Seong Hoon, Son, Woo Sung, Kim, Dae-Duk, Shin, Young Kee
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 23.05.2014; Public Library of Science (PLoS)
• Subjects: Agglomeration; Amino acids; Animals; Binding Sites; Biochemistry; Biological activity; Biology and Life Sciences; Biophysical Phenomena; Dihydrofolate reductase; Fourier transforms; Glycosylation; Humans; Interferon; Interferon beta-1a; Interferon-beta - chemistry; Interferon-beta - genetics; Interferon-beta - metabolism; Interferon-beta - pharmacokinetics; Ligands; Monosaccharides - metabolism; Multiple sclerosis; Mutagenesis; Mutagenesis, Site-Directed; Mutation; N-Acetylneuraminic Acid - metabolism; Pharmaceutical sciences; Pharmacokinetics; Pharmacology; Pharmacy; Physical Sciences; Polyethylene glycol; Properties (attributes); Protein Conformation; Protein Stability; Proteins; Rats; Site-directed mutagenesis; Solubility; Stability; Substrate Specificity; South Korea
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0096967

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.