Glycan analysis of monoclonal antibodies secreted in deposition disorders indicates that subsets of plasma cells differentially process IgG glycans.
OBJECTIVE
To compare the glycosylation of polyclonal serum IgG heavy chains in a patient with rheumatoid arthritis (RA) with that of monoclonal serum IgG heavy chains in the same patient during an episode of heavy-chain deposition disease (HCDD), to establish whether glycosylation processing is specific for subsets of B cells.


METHODS
Serum IgG was purified using a HiTrap protein G column. Immunoglobulins were run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, and IgG glycans were isolated from gel bands and fluorescently labeled. Glycans were analyzed by normal-phase high-performance liquid chromatography and by liquid chromatography-electrospray ionization-mass spectrometry.


RESULTS
The glycosylation of serum immunoglobulins from a patient with seronegative RA and HCDD was analyzed. The predominant immunoglobulin was a truncated glycosylated gamma3 heavy chain, and a small amount of polyclonal IgG was also present. The glycan profile showed that the monoclonal gamma3 heavy chain contained fully galactosylated biantennary glycans with significantly less fucose but more sialic acid than in IgG3 from healthy controls. In contrast, the polyclonal IgG showed an RA-like profile, with a predominance of fucosylated biantennary glycans and low levels of galactosylation. The glycan profile of serum IgG obtained from the same patient during disease remission resembled a typical RA profile.


CONCLUSIONS
These data indicate that different types of B cells process a particular set of IgG glycoforms.
Omtvedt
Lone A
LA
Department of Molecular Biosciences, University of Oslo, Postboks 1041 Blindern, 0316 Oslo, Norway. lone_omtvedt@hotmail.com
Royle
Louise
L
Husby
Gunnar
G
Sletten
Knut
K
Radcliffe
Catherine M
CM
Harvey
David J
DJ
Dwek
Raymond A
RA
Rudd
Pauline M
PM
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D001172:Arthritis, Rheumatoid / Q000276:immunology / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006031:Glycosylation; D006362:Heavy Chain Disease / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D007143:Immunoglobulin Heavy Chains / Q000276:immunology / Q000378:metabolism; D008875:Middle Aged; D010950:Plasma Cells / Q000276:immunology / Q000378:metabolism*; D011134:Polysaccharides / Q000276:immunology / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D011134:Polysaccharides