Here, we present a recently released technical note that will be of interest to scientists engaged in glycoprotein characterization and biopharmaceutical companies designing monosaccharide assays for therapeutic glycoproteins. This second group is growing as many therapeutic glycoproteins are losing patent protection thereby opening up the market to biosimilars or biobetters (both are generic-like products).
Technical Note Glycoprotein Monosaccharide Analysis Using HPAE-PAD with Eluent Generation, (downloadable PDF) is intended to serve as a guide for validating a high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD)-based method for monosaccharide composition analysis. The method uses our latest ion chromatography (IC) system with eluent generation (Thermo Scientific Dionex ICS-5000 Ion Chromatography system), carbohydrate analytical column (Thermo Scientific Dionex CarboPac PA20 Analytical) and our gold disposable electrodes (Gold on PTFE Disposable Electrode PTFE) to improve application ruggedness.
The technical note demonstrates how to determine the monosaccharide composition of both lightly glycosylated (that is, low amounts of carbohydrate) and heavily glycosylated glycoproteins after acid hydrolysis. It also describes what the analyst needs to consider when designing the experiments necessary for determining the monosaccharide composition of a glycoprotein by HPAE-PAD, and the value of using eluent generation and PTFE-AU disposable working electrodes for these determinations.
The method benefits can be summarized as follows:
- The value of HPAE-PAD lies in the fact that it requires less sample preparation than other techniques of monosaccharide analysis and with eluent generation requires no tedious eluent preparation. Thus, eluent generation yields not only convenience, but also high reproducibility.
- Disposable Au on PTFE electrodes have longer lifetimes and can operate at higher hydroxide concentrations compared to earlier-generation disposable working electrodes. They also provide greater system-to-system reproducibility and require no laborious electrode polishing.
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