High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD)

HPAEC-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 HPAEC-PAD analysis.

Glycoproteins Analysis Based on HPAEC-PAD

HPAEC-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, HPAEC-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 that 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, HPAEC-PAD has been used for the initial investigation of a glycoprotein and routine assays of recombinant therapeutic glycoproteins. For example, HPAEC-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 HPAEC-PAD technique has been compared with other chromatographic methods requiring labeling of the sialic acids, showing comparable results. HPAEC-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 HPAEC-PAD monosaccharide composition analysis of bovine fetuin (a, b) and human serum IgG (c, d) (Rohrer, 2013).

HPAEC-PAD Analysis Services

With the years of improvements, our HPAEC-PAD technology has become a sensitive and rapid analysis tool for Glycan Profiling. Based on HPAEC-PAD technique, the monosaccharide composition can be determined. Besides, 13 monosaccharides (such as glucose, fructose, rhamnose, arabinose, fucose, galactose, mannose, ribose, xylose, glucuronic acid, galacturonic acid, mannuronic acid, and guluronic acid), glucosamine, galactosamine, sialic acid, and M-6-P can be quantified by HPAEC-PAD. Our scientists, with extensive experience in HPAEC-PAD analysis, can assist you to design a suitable protocol for glycan profiling.

Features of Our Services Including but Not Limited to:

• Experienced expert team
• Advanced technique platforms
• Providing accurate and comprehensive analysis data as quickly as possible
• High quality and low price services

Creative Biolabs has accumulated rich experience in HPAEC-PAD analysis. We are very confident to offer high-quality and comprehensive services of HPAEC-PAD 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.

Reference

1. Rohrer, J. S.; et al. High-performance anion-exchange chromatography with pulsed amperometric detection for carbohydrate analysis of glycoproteins. Biochemistry. 2013, 78(7):697-709.

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