Characterization of a novel modification on IgG2 light chain: Evidence for the presence of O-linked mannosylation

Abstract This paper describes the analysis of a novel modification identified on the light chain of a recombinant IgG2 antibody. This modification, a +162 Da adduct, suggestive of a single hexose addition, was observed by mass analysis of the reduced molecule. The modification was located on residue serine 66 of the light chain by investigation with LC–MS peptide mapping, mass spectrometry and N-terminal sequencing techniques. Location of the adduct on serine pointed the investigation toward O-linked glycosylation. Identification of the hexose residue was deduced from its elimination by action of α-mannosidase, providing evidence for the presence of an O -mannosylated light chain. This type of modification in the glycosylation profile of antibodies, to our knowledge, has not been reported for human IgG molecules.