This technical note reports an HPAE-PAD method to profile IgG N-linked oligosaccharides and N-linked high-mannose type oligosaccharides that are atypical for human polyclonal IgG but sometimes found on mAbs. This HPAE-PAD method is orthogonal to other oligosaccharide methods applied to IgG. More information on the development and application of the separation described in the technical note can be found in Rohrer, et al. as referenced in the document.

In this work, the oligo- and polysaccharide distribution of three different inulin samples has been characterized by HPAE-PAD. In addition, HPAE-PAD profiling was demonstrated as a method to determine FOS content in prebiotic samples and variation in prebiotics on addition of probiotics. We used Inulin-FOS and chicory root inulin as prebiotic samples. For probiotics, we used Yakult drink, a well known probiotic dairy product. In this method, separation of individual oligosaccharides in FOS was achieved on a Dionex CarboPac PA200 column. An ICS-6000 can be used for this application.

In this study, a method for quantification of six human milk oligosaccharides, namely 2'-fucosyllactose, 3`-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lacto-N-tetraose, and lacto-N-neotetraose in breast milk has been developed and validated. The method is based on the HPAE-PAD technique for carbohydrate determinations without sample derivatization. With an on-line sample cleanup using a polymeric reversed-phase column, a “dilute-and-shoot” sample preparation scheme was applied, which significantly reduces sample handling time and therefore increases sample throughput.

This technical note describes the use of exoglycosidases to obtain structural information about N-linked oligosaccharides commonly found in mammalian glycoproteins. HPAE-PAD is used to confirm the identity of the released monosaccharides, as well as the remaining di-, tri,- or oligosaccharides. Empirical rules used to predict oligosaccharide elution order are included in the last section of this technical note.

This technical note describes the use of exoglycosidases to obtain structural information about N-linked oligosaccharides commonly found in mammalian glycoproteins. HPAE-PAD is used to confirm the identity of the released monosaccharides, as well as the remaining di-, tri,- or oligosaccharides. Empirical rules used to predict oligosaccharide elution order are included in the last section of this technical note.