Analyzing Carbohydrates by HPAE-PAD Ion Chromatography
No need for derivatization or a chromophore
Carbohydrates are carbon sources essential for cell growth and product synthesis, and they play a vital role in biological functions such as cellular communication, gene expression, immunology, and organism defense mechanisms. Characterization of glycoproteins routinely involves carbohydrate analysis, and changes in protein glycosylation are often studied in cancer research to identify potential biomarkers. There are over 30 approved glycoprotein-based biodrugs on the market, and the number is increasing rapidly.
Carbohydrates are difficult to analyze using common chromatography and detection methods. They are very polar compounds, exhibit similar structural characteristics, and lack a suitable chromophore. High-Performance Anion-Exchange Chromatography (HPAE) chromatography takes advantage of the weakly acidic nature of carbohydrates for highly selective separations at high pH using strong anion exchange stationary phases. High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD) does not require modification of glycans before analysis, and specificity is ensured by the separation and the electrochemical detection that has been optimized for carbohydrate analysis.
The U.S. FDA and the European Medicines Agency have increased pressure on biopharmaceutical manufacturers to demonstrate satisfactory programs for understanding, measuring, and controlling glycosylation in glycoprotein-based drugs.