Chemoenzymatic Fc glycosylation via engineered aldehyde tags.
Bioconjug Chem
; 25(4): 788-95, 2014 Apr 16.
Article
em En
| MEDLINE
| ID: mdl-24702330
ABSTRACT
Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. One approach toward constructing such molecules involves chemical glycosylation of aldehyde-tagged proteins. Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Fragmentos Fc das Imunoglobulinas
/
Engenharia de Proteínas
/
Manosil-Glicoproteína Endo-beta-N-Acetilglucosaminidase
/
Aldeídos
Limite:
Animals
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article