title: Five-minute glycoprotein sialic acid determination by high-performance anion exchange chromatography with pulsed amperometric detection.
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.
authors.affiliation: Dionex Products, Thermo Fisher Scientific, Sunnyvale, CA 94085, USA.
authors.forename: Deanna C
authors.initials: DC
authors.lastname: Hurum
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Rohrer
mesh_terms: D000818:Animals; D002138:Calibration / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D057166:High-Throughput Screening Assays; D006801:Humans; D008322:Mammals; D010655:Phenylenediamines / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012794:Sialic Acids / Q000032:analysis* / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: D006023:Glycoproteins; D010655:Phenylenediamines; D012794:Sialic Acids; C058886:1,2-diamino-4,5-methylenedioxybenzene