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HPAE-PAD-Rapid Glycoprotein Sialic Acid Determination by High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detectio.pdf |
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前言:
In this work, a fast (<5 min) and direct chromatographic method for sialic acid determination in therapeutic glycoproteins is developed.
Methods: Determination of two sialic acids,N-acetylneuraminic (Neu5Ac) and N-glycolylneuraminic (Neu5Gc), is performed in two steps. Acid hydrolysis is used to release the sialic acids, which is then followed by a high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) method to determine the amounts of Neu5Ac and Neu5Gc in glycoprotein samples.
仪器:Thermo Scientific Dionex ICS-3000 or 5000 ion chromatography system
结论:
Five model glycoproteins were evaluated for sialic acid content: calf fetuin, bovine apo-transferrin (b. apo-transferrin), human transferrin (h. transferrin), sheep α1-acid glycoprotein (s. AGP), and human α1-acid glycoprotein (h. AGP). The direct HPAE-PAD analyte determination makes this method appropriate for rapid glycoprotein sample screening and eliminates labeling steps common in other methods.