期刊
BIOCONJUGATE CHEMISTRY
卷 20, 期 6, 页码 1228-1236出版社
AMER CHEMICAL SOC
DOI: 10.1021/bc900103p
关键词
-
类别
资金
- National Cancer Institute, National Institutes of Health [N01-CO-12400]
- Intramural Research Program, Center for Cancer Research, National Cancer Institute, NIH
The Fc N-glycan chains of four therapeutic monoclonal antibodies (mAbs), namely, Avastin, Rituxan, Remicade, and Herceptin, released by PNGase F, show by MALDI analysis that these biantermary N-glycans are a mixture of GO, G 1, and G2 glycoforms. The GO glycoform has no galactose on the terminal GlcNAc residues, and the G I and G2 glycoforms have one or two terminal galactose residues, respectively, while no N-glycan with terminal sialic acid residue is observed. We show here that under native conditions we can convert the N-glycans of these mAbs to a homogeneous population of GO glycoform using beta 1,4 galactosidase from Streptococcus pneumoniae. The GO glycoforms of mAbs can be galactosylated with a modified galactose having a chemical handle at the C2 position, such as ketone or azide, using a mutant beta 1,4-galactosyltransferase (beta 1,4Gal-T1-Y289L). The addition of the modified galactose at a specific glycan residue of a mAb permits the coupling of a biomolecule that carries an orthogonal reactive group. The linking of a biotinylated or a fluorescent dye carrying derivatives selectively occurs with the modified galactose, C2-keto-Gal, at the heavy chain of these mAbs, without altering their antigen binding activities, as shown by indirect enzyme linked immunosorbent assay (ELISA) and fluorescence activated cell sorting (FACS) methods. Our results demonstrate that the linking of cargo molecules to mAbs via glycans could prove to be an invaluable tool for potential drug targeting by immunotherapeutic methods.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据