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Five-minute glycoprotein sialic acid determination by high-performance anion exchange chromatography with pulsed amperometric detection.

Research paper by Deanna C DC Hurum, Jeffrey S JS Rohrer

Indexed on: 30 Aug '11Published on: 30 Aug '11Published in: Analytical Biochemistry: Methods in the Biological Sciences



Abstract

Glycoprotein sialylation analysis is a common analytical step in characterizing biotherapeutic products and expression experiments to optimize production. In this article, a high-throughput (5-min) high-performance anion exchange chromatography with pulsed amperometric detection (HPAE-PAD)-based analytical method for glycoprotein sialic acid determination is described. Results from this method are compared with both published HPAE-PAD and 1,2-diamino-4,5-methylenedioxybenzene (DMB) derivatization followed by ultra high-performance liquid chromatography fluorescence detection (UHPLC-FLD) assays. The quantified sialic acid amounts agree with prior HPAE-PAD analyses within replicate error and with UHPLC-FLD within an average of 24%, which are equivalent results based on assay reproducibility.