One-pot enzymatic glycan remodeling of a therapeutic monoclonal antibody by endoglycosidase S (Endo-S) from Streptococcus pyogenes

Xin Tong, Tiezheng Li, Jared Orwenyo, Christian Toonstra, Lai Xi Wang

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

A facile, one-pot enzymatic glycan remodeling of antibody rituximab to produce homogeneous high-mannose and hybrid type antibody glycoforms is described. This method was based on the unique substrate specificity of the endoglycosidase S (Endo-S) from Streptococcus pyogenes. While Endo-S efficiently hydrolyzes the bi-antennary complex type IgG Fc N-glycans, we found that Endo-S did not hydrolyze the “ground state” high-mannose or hybrid glycoforms, and only slowly hydrolyzed the highly activated high-mannose or hybrid N-glycan oxazolines. Moreover, we found that wild-type Endo-S could efficiently use high-mannose or hybrid glycan oxazolines for transglycosylation without product hydrolysis. The combination of the remarkable difference in substrate specificity of Endo-S allows the deglycosylation of heterogeneous rituximab and the transglycosylation with glycan oxazoline to take place in one-pot without the need of isolating the deglycosylated intermediate or changing the enzyme to afford the high-mannose type, hybrid type, and some selectively modified truncated form of antibody glycoforms.

Original languageEnglish (US)
Pages (from-to)1347-1355
Number of pages9
JournalBioorganic and Medicinal Chemistry
Volume26
Issue number7
DOIs
StatePublished - Apr 1 2018

Bibliographical note

Funding Information:
We thank other members of the Wang lab for technical assistance and helpful discussions. This work was supported by the US National Institutes of Health (NIH grant R01 GM096973 ).

Funding Information:
We thank other members of the Wang lab for technical assistance and helpful discussions. This work was supported by the US National Institutes of Health (NIH grant R01 GM096973).

Keywords

  • Chemoenzymatic synthesis
  • Endo S
  • Endoglycosidase
  • Fc glycosylation
  • Glycoengineering
  • Monoclonal antibody
  • Rituximab
  • Transglycosylation

Fingerprint Dive into the research topics of 'One-pot enzymatic glycan remodeling of a therapeutic monoclonal antibody by endoglycosidase S (Endo-S) from Streptococcus pyogenes'. Together they form a unique fingerprint.

Cite this