Glycoengineering of Interferon-beta 1a Improves Its Biophysical and Pharmacokinetic Properties

Abstract

The purpose of this study was to develop a biobetter version of recombinant human interferon-beta 1a (rhIFN-beta 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-beta 1a via site-directed mutagenesis. Glycoengineering of rhIFN-beta 1a resulted in a new molecular entity, termed R27T, which was defined as a rhIFN-beta 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-beta 1a. Glycoengineering had no effect on rhIFN-beta 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-beta could be a biobetter version of rhIFN-beta 1a with a potential for use as a drug against multiple sclerosis.