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Cited by 112Glycosylation and Glycoproteins ResearchCarbohydrate Chemistry and SynthesisViral Infectious Diseases and Gene Expression in Insects

Site-specific enzymatic polysialylation of therapeutic proteins using bacterial enzymes

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Lindhout, Theresa, Iqbal, Umar, Willis, Lisa M., Reid, Anne N., Li, Jianjun, Liu, Xin, Moreno, Maria, Wakarchuk, Warren W.·2011·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.1019266108PMC PMC3088639
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Abstract

The posttranslational modification of therapeutic proteins with terminal sialic acids is one means of improving their circulating half-life, thereby improving their efficiency. We have developed a two-step in vitro enzymatic modification of glycoproteins, which has previously only been achieved by chemical means [Gregoriadis G, Jain S, Papaioannou I, Laing P (2005) Int J Pharm 300:125–130). This two-step procedure uses the Campylobacter jejuni Cst-II α2,8-sialyltransferase to provide a primer on N-linked glycans, followed by polysialylation using the Neisseria meningitidis α2,8-polysialyltransferase. Here, we have demonstrated the ability of this system to modify three glycoproteins with varying N-linked glycan compositions: the human therapeutic proteins alpha-1-antitrypsin (A1AT) and factor IX, as well as bovine fetuin. The chain length of the polysialic acid addition was optimized by controlling reaction conditions. After demonstrating the ability of this system to modify a variety of proteins, the effect of polysialylation on the activity and serum half-life of A1AT was examined. The polysialylation of A1AT did not adversely affect its in vitro inhibition activity against human neutrophil elastase. The polysialylation of A1AT resulted in a significantly improved pharmacokinetic profile when the modified proteins were injected into CD-1 mice. Together, these results suggest that polysialylated A1AT may be useful for improved augmentation therapy for patients with a deficiency in this protein and that this modification may be applied to other therapeutic proteins.

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  • Cst-II allows efficient polysialylation by the bacterial polysialyltransferase on recombinant alpha-1-antitrypsin in vitro and this results in a significantly longer half-life in mice, without affecting its specific activity.

    “Cst-II, a bacterial sialyltransferase that can catalyze α2,8 sialylation, allows efficient polysialylation by the bacterial polysialyltransferase on recombinant alpha-1-antitrypsin in vitro and this results in a significantly longer half-life in mice, without affecting its specific activity (Lindhout et al., 2011).”

    From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Lindhout et al., 2011

    The Abstract describes Cst-II priming A1AT for in vitro polysialylation, and the Results report a longer terminal half-life in mice without adversely affecting its in vitro inhibitory activity. However, the Discussion says the A1AT used was derived from human plasma, not recombinant A1AT. (LLM, conf=0.90)

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Authors
Lindhout, T., Iqbal, U., Willis, L. M., Reid, A. N., Li, J., Liu, X., Moreno, M., & Wakarchuk, W. W.
Affiliations
  • University of Guelph (CA)
  • Institute for Biological Sciences (CA)
  • Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6
Corresponding author
Wakarchuk, Warren W.
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2011-04-18
Volume
108
Issue
18
Pages
7397-7402
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.1019266108
PMC
PMC3088639
OpenAlex
W2009125126
Cited by
112
References
0
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FWCI
3.89
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • NPARC (submittedVersion)
Funders
Canadian Institutes of Health Research (MOP84272)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
Keywords
polysialylation, sialic acid, therapeutic proteins, glycoproteins, N-glycans, alpha-1-antitrypsin, factor IX, fetuin-A, pharmacokinetics, serum half-life, post-translational modification, neutrophil elastase inhibition, bacterial enzymes
MeSH terms
alpha-Fetoproteins, alpha 1-Antitrypsin, Animals, Cattle, Chromatography, Electrophoresis, Polyacrylamide Gel, Factor IX, Fluorescence, Glycoproteins, Humans, Neisseria meningitidis, Protein Processing, Post-Translational, Sialyltransferases, Mass Spectrometry, Drug Design, Campylobacter jejuni, Mice, In Vitro Techniques

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