A study of immunoglobulin G glycosylation in monoclonal and polyclonal species by electrospray and matrix-assisted laser desorption/ionization mass spectrometry

被引:36
作者
Saba, JA
Kunkel, JP
Jan, DCH
Ens, WE
Standing, KG
Butler, M
Jamieson, JC
Perreault, H
机构
[1] Univ Manitoba, Dept Chem, Winnipeg, MB R3T 2N2, Canada
[2] Univ Manitoba, Dept Microbiol, Winnipeg, MB R3T 2N2, Canada
[3] Univ Manitoba, Dept Phys & Astron, Winnipeg, MB R3T 2N2, Canada
关键词
monoclonal antibody; glycosylation; glycoprotein; FACE; ESI-MS; HPLC; MALDI-TOF-MS; sugar labeling; HPAEC-PAD;
D O I
10.1006/abio.2002.5651
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
N-linked oligosaccharides were released from human and bovine polyclonal immunoglobulin G (IgG) obtained from commercial sources and also from a monoclonal IgG(1) secreted by murine B-lymphocyte hybridoma cells (CC9C10) grown under different serum-free conditions. These conditions differed according to their steady-state dissolved oxygen concentrations. This work is based on a previous quantitative study where released glycans were characterized by fluorophore-assisted carbohydrate electrophoresis (FACE) and high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) (J. P. Kunkel, D. C. H. Jan, J. C. Jamieson, and M. Butler, 1998, J. Biotechnol, 62, 55-71). In the present article, peptide-N-glycosidase F-released glycans from different species of polyclonal IgG and murine monoclonal IgG were characterized qualitatively by high-performance liquid chromatography (HPLC) coupled to electrospray ionization mass spectrometry (ESI-MS). The glycans were also analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The MALDI mass spectrometer used allowed acquisition of MS and tandem MS data, which were useful in structural investigations at a more detailed level than allowed by FACE and HPAEC-PAD. Predominant N-linked structures, as determined by all techniques, were core-fucosyl asialyl biantennary chains with varying galactosylation. Minor amounts of afucosyl, bisected, and monosialyl oligosaccharides were also detected. In contrast to FACE and HPAEC-PAD, MALDI-double quadrupole/time-of-flight MS and HPLC/ESI-MS also detected low-abundance high-mannose and hybrid structures in some of the species under investigation. (C) 2002 Elsevier Science (USA).
引用
收藏
页码:16 / 31
页数:16
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