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Glycoconjugation of polypeptides using oligosaccharyltransferases

A polypeptide and glycosyl technology, applied in the field of polypeptide modification, can solve the problems of unsuitable attachment of modified groups and reduced biological activity of polypeptides.

Inactive Publication Date: 2011-04-27
蔚所番有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Additionally, existing glycosylation sequences may not be suitable for attachment of modifying groups
Such modifications may, for example, cause an undesired decrease in the biological activity of the modified polypeptide

Method used

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  • Glycoconjugation of polypeptides using oligosaccharyltransferases
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  • Glycoconjugation of polypeptides using oligosaccharyltransferases

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[0032] I. abbreviation

[0033] PEG, poly(ethylene glycol); m-PEG, methoxy-poly(ethylene glycol); PPG, poly(propylene glycol); m-PPG, methoxy-poly(propylene glycol); Fuc, fucose or Fucosyl; Gal, galactose or galactosyl; GalNAc, N-acetylgalactosamine or N-acetylgalactosamine; Glc, glucose or glucosyl; GlcNAc, N-acetylglucosamine or N-acetyl Glucosamine; Man, mannose or mannosyl; ManAc, mannosamine acetate or mannosamine acetate; Sia, sialic acid or sialyl; and NeuAc, N-acetylceramide or N-acetylceramide base.

[0034] II. definition

[0035] Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature and laboratory practices in cell culture, molecular genetics, organic and nucleic acid chemistry and hybridization used herein are those well known and commonly employed in the art. Standard techniques are used ...

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Abstract

The current invention provides polypeptides and polypeptide conjugates that include an exogenous N-linked glycosylation sequence. The N-linked glycosylation sequence is preferably a substrate for an oligosaccharyltransferase (e.g., bacterial PgIB), which can catalyze the transfer of a glycosyl moiety from a lipid-bound glycosyl donor molecule (e.g., a lipid-pyrophosphate-linked glycosyl moiety) to an asparagine (N) residue of the glycosylation sequence. In one example, the asparagine residue is part of an exogenous N-linked glycosylation sequence of the invention. The invention further provides methods of making the polypeptide conjugates that include contacting a polypeptide having an N-linked glycosylation sequence of the invention and a lipid-pyrophosphate-linked glycosyl moiety (or phospholipid-linked glycosyl moiety) in the presence of an oligosaccharyltransferase under conditions sufficient for the enzyme to transfer the glycosyl moiety to an asparagine residue of the N-linked glycosylation sequence. Exemplary glycosyl moieties that can be conjugated to the glycosylation sequence include GlcNAc, GlcNH, bacillosamine, 6-hydroybacillosamine, GalNAc, GaINH, GlcNAc-GlcNAc, GlcNAc-GlcNH, GlcNAc-Gal, GlcNAc-GlcNAc-Gal-Sia, GlcNAc-Gal-Sia, GlcNAc-GlcNAc-Man, and GlcNAc-GlcNAc-Man(Man)2. The transferred glycosyl moiety is optionally modified with a modifying group, such as a polymer (e.g., PEG). In one example, the modified glycosyl moiety is a GIcNAc or a sialic acid moiety.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Patent Application No. 61 / 019,805, filed January 8, 2008, the contents of which are hereby incorporated by reference in their entirety for all purposes. field of invention [0003] The present invention relates to the field of polypeptide modification by glycosylation. In particular, the invention relates to methods for preparing glycosylated polypeptides using short enzyme-recognized N-linked glycosylation sequences. Background of the invention [0004] The administration of glycosylated and non-glycosylated polypeptides to elicit specific physiological responses is well known in the medical arts. For example, purified and recombinant human growth hormone (hGH) is used to treat conditions and diseases associated with hGH deficiency, such as dwarfism in children. Other examples involve interferon, which has known antiviral activity, and granulocyte colony-stimulat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/107
CPCC12N9/644C12Y304/21022C12Y304/21021C12N9/6437A61K47/48046C12Y204/01119A61K38/00C07K14/48C07K14/755C07K14/51C07K14/50A61K47/48215A61K47/48092C12Y204/99018A61K47/60A61K47/543A61K47/549
Inventor S·德弗里斯
Owner 蔚所番有限公司
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