Analysis of 3-(acetylamino)-6-aminoacridine-derivatized oligosaccharides from recombinant monoclonal antibodies by liquid chromatography-mass spectrometry.

Glycosylation has been established as playing a pivotal role in various aspects of recombinant monoclonal antibodies (MAbs), ranging from pharmacokinetics to enhancement of effector function. Consequently, characterization of these oligosaccharides is of great importance and requires sensitive analytical techniques. Here we present a method for the rapid elucidation of 3-(acetylamino)-6-aminoacridine-labeled N-glycans present on MAbs using liquid chromatography-mass spectrometry. The technique uses the benefits of ultra-performance liquid chromatography systems in conjunction with small-particle-size amide columns capable of generating a fluorescence glycan profile of a MAb in 30 min, reducing the current run time by a factor of 6. The method is also compatible with online electrospray mass spectrometry, permitting the identification of glycans present. Overall, this strategy allows the confident determination of N-glycans present on recombinant MAbs in a significantly reduced amount of time.