Enhancement of therapeutic protein in vivo activities through glycoengineering

  title={Enhancement of therapeutic protein in vivo activities through glycoengineering},
  author={Steven G. Elliott and Tony Lorenzini and Sheilah J. Asher and Kenneth H. Aoki and David W. Brankow and Lynette Buck and Leigh A. Busse and David Chang and Janis Fuller and James Grant and Natasha Hernday and Martha Hokum and Sylvia Hu and Andrew Knudten and Nancy J Levin and Renee Komorowski and F Bachmann Martin and Rachell Navarro and Timothy D. Osslund and Gary Rogers and Norma Rogers and Geri Trail and Joan C. Egrie},
  journal={Nature Biotechnology},
Delivery of protein therapeutics often requires frequent injections because of low activity or rapid clearance, thereby placing a burden on patients and caregivers. Using glycoengineering, we have increased and prolonged the activity of proteins, thus allowing reduced frequency of administration. Glycosylation analogs with new N-linked glycosylation consensus sequences introduced into the protein were screened for the presence of additional N-linked carbohydrates and retention of in vitro… 
Glycoengineering: the effect of glycosylation on the properties of therapeutic proteins.
The discovery of darbepoetin alfa (DA), a hyperglycosylated analogue of erythropoietin that contains two additional N-linked carbohydrates, a threefold increase in serum half-life and increased in vivo activity compared to recombinant human ery Anthropo-EPO, is discussed.
Glycosylation of Therapeutic Proteins
An account of the effects that glycosylation has on the therapeutic efficacy of protein drugs is provided and the current understanding of the mechanisms by which glyCosylation leads to such effects are described.
Optimization of physicochemical and pharmacological properties of peptide drugs by glycosylation.
Various synthetic approaches to prepare N- and O-glycopeptides bearing simple monosaccharides as a tool to improve peptide therapeutic efficacy by glycosylation are described.
Therapeutic proteins: Glycoengineered to last longer
  • S. Farley
  • Biology
    Nature Reviews Drug Discovery
  • 2003
This success, and further experiments reported by Elliott and colleagues showing that glycosylation analogues of leptin and Mpl ligand also had increased in vivo activity and duration of action, indicates that glycoengineering could be a generally applicable strategy for improving therapeutic proteins.
Chemical (neo)glycosylation of biological drugs.
Glycosylated enfuvirtide: a long-lasting glycopeptide with potent anti-HIV activity.
It is reported here that the pharmacokinetics of an anti-HIV peptide drug enfuvirtide (ENF) can be dramatically improved by a chemical glycosylation approach, and this approach may be useful for the development of novel protein and peptide drugs with enhanced pharmaceutical properties.


Role of glycosylation on the secretion and biological activity of erythropoietin.
Removal of any of the N-glycosylation sites reduced the in vivo but not the in vitro biological activity of the EPO molecule, which indicates that the O-linked carbohydrate is not essential for activity.
Development and characterization of novel erythropoiesis stimulating protein (NESP)
Darbepoetin alfa (novel erythropoiesis stimulating protein, NESP, ARANESPTM, Amgen Inc, Thousand Oaks, CA), which was engineered to contain 5 N-linked carbohydrate chains, is currently being evaluated in human clinical trials for treatment of anaemia and reduction in its incidence.
Prolonged in vivo anticoagulant activity of a hirudin–albumin fusion protein secreted from Pichia pastoris
The results suggest that HLAH6 exhibits not only delayed clearance, but also prolonged biological activity in vivo compared with unfused hirudin, which inhibited clot-bound thrombin.
The uses and properties of PEG-linked proteins.
PEG-modified cytokines have been constructed and one of the conjugates, PEG- modified granulocyte-macrophage colony-stimulating factor, showed dissociation of two biological properties, which may open new horizons to the application of PEGylation technology.
An overview of the efficacy and safety of novel erythropoiesis stimulating protein (NESP).
  • I. Macdougall
  • Medicine, Biology
    Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association
  • 2001
Novo erythropoiesis stimulating protein is well tolerated, adverse effects are similar to those seen with rHuEPO, and no antibodies have been detected in >1500 patients exposed to NESP thus far.
Mapping of the active site of recombinant human erythropoietin.
Recombinant human erythropoietin variants found to be important for bioactivity were divided into two groups according to the differential effects on EPO receptor binding activity and in vitro biologic activity, suggesting that rHuEPO has two separate receptor binding sites.
Human erythropoietin dimers with markedly enhanced in vivo activity.
The production of biologically active erythropoietin dimers and trimers by chemical crosslinking of the conventional monomeric form exhibited >26-fold higher activity in vivo than did the monomers and were very effective after only one dose.
Structural requirements for addition of O-linked carbohydrate to recombinant erythropoietin.
It is found that glycosylation is less efficient when rEPO is improperly folded and that prolines at -1 and +1 relative to the O-glycosylations site enhance glycosolation.