WebIn this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a ... WebHILIC Overview. HILIC can be described as a variation of reversed phase chromatography performed using a polar stationary phase. The mobile phase employed is highly organic in …
USP Reference Standard Monoclonal Antibodies: Tools to Verify Glycan …
WebGlycosylation of biotherapeutics can impact drug safety, efficacy, and stability, therefore it is often designated as a critical quality attribute (CQA) in biopharmaceutical development. 1 Given its sensitivity to upstream processing conditions, glycosylation also serves as a key indicator of process stability and robustness. 2 In addition to the … WebFollowing HILIC-FLD analysis, routine glycan assignment is facilitated by Glucose Unit (GU) normalization, often through retention time calibration using a labelled dextran ladder. Through creation of a relevant GU retention library, glycan identification can be performed routinely, without the need for prior mass spectrometric analysis. theory j manager
LC-MS/MS Separation of 38 Amino Acids in HILIC Mode
WebHILIC-UHPLC analysis of labeled N-glycans results in separation of the major glycans for both MabThera (Figure 2A) and Enbrel (Figure 2B). The fluorescence intensities of equivalent quantities of glycans loaded on column varies by the dye used for labeling (Figure 2 A inset). While retention time of labeled N-glycans is WebNov 26, 2024 · HILIC separation of glycans was performed on a 1290 Infinity II UHPLC system (Agilent, Waldbronn, Germany) optimized for ultra-low dispersion, operated at a flow rate of 0.6 mL min −1 and a temperature of 55 °C. Fluorescence detection of 2-AA-labeled N-glycans was carried out with an excitation wavelength of 350 nm and emission … WebHILIC was performed on 1 µL aqueous injections of 5 mg/mL trastuzumab using an ACQUITY UPLC Glycoprotein BEH Amide 300Å 1.7 µm Column heated to 80˚C. Sample … shrubs for privacy fence