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Improvement of in vitro stability and pharmacokinetics of hIFN-α by fusing the carboxyl-terminal peptide of hCG β-subunit

Paper ID Volume ID Publish Year Pages File Format Full-Text
22717 43373 2016 12 PDF Available
Title
Improvement of in vitro stability and pharmacokinetics of hIFN-α by fusing the carboxyl-terminal peptide of hCG β-subunit
Abstract

•A strategy based on the fusion of carboxyl-terminal peptide (CTP) of hCG was applied to hIFN-α2b.•CTP remarkably improved the rhIFN-α2b stability after thermal and plasma protease treatment.•CTP fusion significantly enhanced the pharmacokinetic properties of rhIFN-α2b.•CHO-derived O-glycans of CTP resemble hCG glycans found in human normal cells.

Improving in vivo half-life and in vitro stability of protein-based therapeutics is a current challenge for the biopharmaceutical industry. In particular, recombinant human interferon alpha-2b (rhIFN-α2b), which belongs to a group of cytokines extensively used for the treatment of viral diseases and cancers, shows a poor stability in solution and an extremely short plasma half-life which determines a strict therapeutic regimen comprising high and repeated doses. In this work, we have used a strategy based on the fusion of the carboxyl-terminal peptide (CTP) of human chorionic gonadotropin (hCG) β-subunit, bearing four O-linked oligosaccharide recognition sites, to each or both N- and C-terminal ends of rhIFN-α2b. Molecules containing from 5 (CTP-IFN and IFN-CTP) to 9 (CTP-IFN-CTP) O-glycosylation sites were efficiently expressed and secreted to CHO cells supernatants, and exhibited antiviral and antiproliferative bioactivities in vitro. Significant improvements in pharmacokinetics in rats were achieved through this approach, since the doubly CTP-modified IFN variant showed a 10-fold longer elimination half-life and a 19-fold decreased plasma apparent clearance compared to the wild-type cytokine. Moreover, CTP-IFN-CTP demonstrated a significant increase in in vitro thermal resistance and a higher stability against plasma protease inactivation, both features attributed to the stabilizing effects of the O-glycans provided by the CTP moiety. These results constitute the first report that postulates CTP as a tag for improving both the in vitro and in vivo stability of rhIFN-α2b which, in turn, would positively influence its in vivo bioactivity.

Keywords
hIFN-α, human interferon alpha; hCG, human chorionic gonadotropin; CTP, carboxyl-terminal peptide; LH, luteinizing hormone; FSH, follicle-stimulating hormone; TSH, thyrotropin; EPO, erythropoietin; LV, lentiviral; LVP, lentiviral particle; Cmax, maximum p
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Improvement of in vitro stability and pharmacokinetics of hIFN-α by fusing the carboxyl-terminal peptide of hCG β-subunit
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 221, 10 March 2016, Pages 13–24
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us