Glycosylation of human IgG subclass and mouse IgG2b heavy chains secreted by mouse J558L transfectoma cell lines as chimeric antibodies.
Oligosaccharide profiles for monoclonal chimeric mouse-human and mouse IgG2b antibodies produced in the mouse J558L cell line have been determined. These chimeric antibodies share the same mouse light chain and VH region and have a single, complex, oligosaccharide moiety attached to residue 297 of the heavy chain. Since the panel of chimeric antibodies included each of the human IgG subclasses and a set of IgG3 proteins having single residue differences, it was possible to evaluate the contributions of the protein template, the cell line, and the culture conditions to glycosylation. The study shows that the J558L cell line glycosylates human heavy chains with oligosaccharides characteristic of mouse immunoglobulins. This includes a galactose alpha 1-->3 galactose structure not found in human IgG and for which most humans have naturally occurring "anti-Gal" antibodies. The extent of galactosylation was very dependent on the culture conditions and was maximal for still culture. The findings have significance for the production of antibodies for in vivo administration.
Lund
J
J
Department of Immunology, University of Birmingham Medical School, UK.
Takahashi
N
N
Hindley
S
S
Tyler
R
R
Goodall
M
M
Jefferis
R
R
D000818:Animals; D002240:Carbohydrate Sequence; D006031:Glycosylation; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism*; D007074:Immunoglobulin G / Q000145:classification / Q000378:metabolism*; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D014162:Transfection
D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D009844:Oligosaccharides; D011993:Recombinant Fusion Proteins