Stable isotope labeling for profiling of IgG Fc glycosylation


Iva Tomalová


Mass spectrometric approach together with stable isotope labeling of glycopeptides was applied to
monitor changes in glycosylation profile of Fc region of immunoglobulin G. Succinic anhydride was
used as a label as it is known to bind easily to primary amino groups present in all peptides and
does not affect the glycan portion during the labeling reaction. A simple sample preparation
protocol consisting of an in-solution trypsin digestion, a 60 minutes “one-pot” labeling procedure
and a solid phase extraction was established. Analysis of the labeled glycopeptides was finally
done by nano-LC-MALDI-MS, nano-LC-ESI-MS or by a direct MALDI-MS spotting.