HPAE-PAD – A very useful tool for glycan analysis

by | Dec 13, 2017

High-performance anion-exchange chromatography with pulsed amperometric detection - a method for monosaccharides, sialic acids, and other carbohydrates analysis

Looking for analytical advantage with your glycan and glycosylation analysis? If so, then high-performance anion-exchange chromatography with pulsed amperometric detection may well be worth a look...

A large number of innovator biotherapeutics are currently losing patent protection, resulting in an increased interest to develop biosimilar therapeutic proteins. A key step to proving biosimilarity is to ensure proper N-glycosylations, because variations in glycosylation can affect the efficacy and safety of therapeutic proteins, influencing their biological activity, serum half-life, and immunogenicity. Monosaccharides, including sialic acid composition analysis can detect variations in glycosylation, and support quality control for process development and manufacturing procedures. As a result, proteins are routinely analysed to determine their monosaccharides and sialic acids amount and identity.

High-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) is a well-established method for carbohydrates analysis. HPAE-PAD separates carbohydrates with specific interactions between the hydroxyl and carboxyl groups of carbohydrates based on charge, size, composition, isomers, and linkages. Therefore, HPAE-PAD is the best method for monosaccharides, sialic acids, and other carbohydrates analysis. Why?

  • Direct detection - no sample derivatization required saving analyst time, expense and exposure to hazardous chemicals
  • Ultra-selective, sensitive detection
  • Fast separations without loss of resolution using Thermo Scientific™ Dionex™ CarboPac™ PA20 Fast columns

Separation Science, in collaboration with Thermo Fisher Scientific, has developed an educational microsite providing learning content, advice and guidance for the transition to modernized pharmacopoeial methods using IC. As part of this microsite, you are able to download a recent publication titled 'Monosaccharide and Sialic Acid Determinations in Biosimilars Using HPAE-PAD', which provides detailed information on the use of HPAE-PAD for glycan analysis. Applications described include:

  • Glycoprotein Monsaccharide Analysis Using HPAE-PAD with Eluent Generation
  • Rapid Screening of Sialic Acid in Glycoproteins by HPAE-PAD
  • Evaluating Protein Glycosylation in Limited-Quantity Samples by HPAE-PAD
  • Sialic Acids in Glycoprotein Hydrolyzates by HPAE-PAD

The comprehensive booklet describes the technology behind HPAE-PAD in great detail, as well as the systems and instruments available. If you have a role in glycan analysis in your company, why not take a look - it could be just the technique you're looking for.

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