HPAE-PAD becomes a popular method that is widely used for the determination of a large variety of carbohydrates with the advantages of high-resolution separations and sensitive detection. Creative Biolabs is a world-leading custom services provider in the field of glycoprotein analysis. Based on HPAE-PAD technique, we have established a useful glycoprotein analysis platform to offer a variety of glycoproteins analysis services.
HPAE-PAD is a well-established technique for glycoprotein analysis as well as for other fields of carbohydrate analysis. It was first applied to glycoprotein analysis in the late 1980s. Currently, HPAE-PAD is widely used for determinations of monosaccharide, sialic acid, mannose-6-phosphate (M-6-P), and oligosaccharide contents of a glycoprotein. This technique can effectively separate carbohydrates which become oxyanions at high pH using anion-exchange chromatography. The separated carbohydrates can be detected by PAD which is a direct and sensitive detection technique. It allows detecting carbohydrates, with excellent signal-to-noise ratios and sensitivities down to subpicomole levels for standard bore and microbore systems and femtomole levels for capillary systems, respectively, without requiring derivatization. For these advantages, HPAE-PAD has been used for the initial investigation of a glycoprotein and routine assays of recombinant therapeutic glycoproteins. For example, HPAE-PAD has been used for the determination of the monosaccharide and sialic acids content of a glycoprotein. The determination of individual sialic acids content in proteins by HPAE-PAD technique have been compared with other chromatographic methods requiring labeling of the sialic acids, showing comparable results. HPAE-PAD is also performed to determine the relative amounts of N-acetylneuraminic acid (Neu5Ac) and Neu5Gc as well as the M-6-P content of a glycoprotein.
Fig. 1 HPAE-PAD monosaccharide composition analysis of bovine fetuin (a, b) and human serum IgG (c, d) (Rohrer, 2013).
With the years of improvements, our HPAE-PAD technology has become a sensitive and rapid analysis tool for Glycan Profiling. Based on HPAE-PAD technique, the monosaccharide composition of a therapeutic protein such as antibody can be determined. Besides, the contents of monosaccharide, sialic acid, M-6-P, and oligosaccharide of a glycoprotein also can be determined. Our scientists, with extensive experience in glycoproteins analysis, can assist you to design the suitable protocol for glycan profiling based on HPAE-PAD technique.
Our expert team has accumulated rich experience in glycoproteins analysis. We are very confident to offer high-quality and comprehensive services of glycoproteins analysis for global clients. With our excellent service, we will offer you not only an accurate analysis result but also our professional technical guidance to facilitate the later project.
If you are interested in our HPAEC-PAD analysis services, please feel free to contact us for more details.