Characterization of Glycosylated Proteins at Subunit Level by HILIC/MS

Hydrophilic interaction chromatography (HILIC) coupled to mass spectrometry (MS) is considered as the reference analytical technique for glycans profiling, especially characterization of glycosylated protein therapeutics such monoclonal antibodies (mAbs) and mAbs-related products. Although HILIC/MS mainly known profile enzymatically released fluorescently labeled N-glycans, recent commercialization new widepore HILIC amide bonded stationary phases packed with sub-2 μm particles has allowed remarkable separations also at subunit level. Here, we describe a simple protocol perform mAb profiling level by HILIC/MS.