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Protein Glycosylation Evaluation in Less Than 10 µg of Protein Samples

Published: Friday, August 23, 2013
Last Updated: Friday, August 23, 2013
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Evaluating protein glycosylation in limited-quantity samples by HPAE-PAD.

Thermo Fisher Scientific has developed a new approach that combines high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) analysis, acid hydrolysis, and enzymatic digestion of protein glycans to determine information about glycan identity and terminal carbohydrate linkage isomers from small amounts of protein-just 0.5–1.6 μg per injection.

Application Note (AN) 1050: Evaluating Protein Glycosylation in Limited-Quantity Samples by HPAE-PAD demonstrates that a suite of HPAE-PAD analyses can be performed with sample quantities as low as 3-10 μg. This direct determination approach saves time and reagent costs.

Cancer research frequently involves studying changes in protein glycosylation to identify important biomarkers such as prostate-specific antigen (PSA).

The AN 1050 methodology uses an ion chromatography system configured for HPAE-PAD to investigate human PSA N-linked glycosylation and to evaluate the potential presence of O-linked glycans with methods optimized to work with small quantities of glycoproteins.

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