Thermo Fisher Scientific has released a technical note (TN) that challenges the common misconception that there is significant variation in high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) peak area response of oligosaccharides. TN 133: HPAE-PAD Peak Area Response of Glycoprotein Oligosaccharides summarizes the peer-reviewed scientific literature concerning this subject. This review concludes that the variation in HPAE-PAD oligosaccharide response with structure is minimal, especially among sets of similar oligosaccharides such as those found on many glycoproteins.
HPAE-PAD is a key technique used for characterizing recombinant therapeutic glycosylation because it can be used for monosaccharide, sialic acid, and oligosaccharide analyses (Asn-linked [N-linked] and Ser/Thr-linked [O-linked] oligosaccharides)—commonly referred to as oligosaccharide profiling. In this review of scientific literature evaluating electrochemical response of glycoprotein-derived and related oligosaccharides, TN 133 shows that the changes of electrochemical response with structure are not extreme. TN 133 then compares HPAE-PAD to other common methods of oligosaccharide profiling, including matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis of oligosaccharides.