Journal
ANALYTICAL BIOCHEMISTRY
Volume 419, Issue 1, Pages 67-69Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2011.08.002
Keywords
Sialic acids; Glycoproteins; Anion exchange; Pulsed amperometric detection; HPAE-PAD; Neu5Ac; Neu5Gc
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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. (C) 2011 Elsevier Inc. All rights reserved.
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