Fractionation of Fab Glycosylated Immunoglobulin G with Concanavalin A Chromatography Unveils New Structural Properties of the Molecule

Overview

Journal Oncotarget

Specialty Oncology

Date 2016 May 5

PMID 27145274

Citations 4

Authors

Tao Huang

Xueling Chen

Huan Gu

Conghui Zhao

Xingmu Liu

Meiling Yan

Xiaodong Deng

Zaiping Zhang

Jiang Gu

Affiliations

Soon will be listed here.

Abstract

Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications.

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