Journal
JOURNAL OF BIOTECHNOLOGY
Volume 146, Issue 1-2, Pages 74-83Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2009.12.020
Keywords
Interferon alpha; Glycoengineering; Antitumor activity; Pharmacokinetics; Cytokine analog
Categories
Funding
- Universidad Nacional del Litoral [5/38, 356,698]
- Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET)
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The type I human interferon alpha (hIFN-alpha) family consists of small proteins that exert a multiplicity of biological actions including antiviral, antiproliferative and immunomodulatory effects. However, though administration of recombinant hIFN-alpha 2b is the current treatment for chronic hepatitis B and C and for some types of cancers, therapy outcomes have not been completely satisfactory. The short serum half-life and rapid clearance of the cytokine accounts for its low in vivo biological activity. Here we describe and characterize a long-acting rhIFN-alpha 2b mutein, 4N-IFN, which has been created by introducing four N-glycosylation sites via site-directed mutagenesis. The hyperglycosylated protein was found to have a 25-fold longer plasma half-life than the non-glycosylated rhIFN-alpha 2b, even greater than the commercial pegylated derivative Intron-A PEG. In addition, glycosylation increased the in vitro stability of the mutein against serum protease inactivation. Interestingly, despite its lower in vitro activity, 4N-IFN showed a markedly enhanced in vivo antitumor activity in human prostate carcinoma implanted in nude mice. MALDI-TOF MS and HPAEC-PAD carbohydrate analyses revealed the presence of high amounts of tetrasialylated (40%) and trisialylated (28%) N-glycan structures, which are consequently responsible for the improved characteristics of the cytokine, making 4N-IFN a new therapeutic candidate for viral and malignant diseases. (C) 2010 Elsevier B.V. All rights reserved.
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