--- title: BioPharmaceutical Characterization Resource Center - Unlabelled Glycan --- [Chromatography Forum](http://www.chromforum.org/) [Analytical Training Solutions](https://www.analytical-training-solutions.com) [日本語](https://www.sepscience.com/japan) [![Separation Science](https://learning.sepscience.com/hubfs/Sepscience_Dec2016/images/-resized-strch-SepSci_WebHeader_v3.png)](https://www.sepscience.com/) [Chromatography Forum](http://www.chromforum.org/) [Analytical Training Solutions](https://www.analytical-training-solutions.com) [日本語](https://blog.sepscience.com/japan) [Sign Up for Newsletter](https://www.sepscience.com/subscribe/) ![HeaderImage_v2](https://learning.sepscience.com/hs-fs/hubfs/HeaderImage_v2.png?width=1280&name=HeaderImage_v2.png)  [![Home](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/Home.png?width=47&name=Home.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-home)  [![PeptideMapping](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/PeptideMapping.png?width=118&name=PeptideMapping.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-peptide-mapping)  [![LabelledGlycan](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/LabelledGlycan.png?width=158&name=LabelledGlycan.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-labelled-glycans)  [![UnlabelledGlycan](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/UnlabelledGlycan.png?width=189&name=UnlabelledGlycan.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-unlabelled-glycan)  [![NativeMass](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/NativeMass.png?width=142&name=NativeMass.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-native-mass-analysis)  [![AggregateAnalysis](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/AggregateAnalysis.png?width=126&name=AggregateAnalysis.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-aggregate-analysis)  [![ChargeVariantAnalysis](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/ChargeVariantAnalysis.png?width=158&name=ChargeVariantAnalysis.png)](https://learning.sepscience.com/biopharmaceutical-characterization-resource-center-charge-variant-analysis) ## Unlabelled Glycan and Monosacharride Analysis Glycosylation analysis is one of the key critical quality attributes of monoclonal antibody–based biotherapeutics. Glycosylation changes can impact a biological drug’s safety, efficacy, clearance, and immunogenicity, making it necessary to accurately detect changes. Although glycans are often derivatized prior to analysis, it is also possible to analyze both N-glycans and O-Glycans in a label-free form, typically using ion chromatography (IC) or liquid chromatography with charged aerosol detection. IC technique is sensitive and selective and resolves glycans according to charge, size, composition, isomers and linkage. Monosaccharide and sialic acid analysis is applicable to all stages of biopharmaceutical development and manufacturing processes including cell line and clone selection, upstream and downstream process development and manufacturing in-process control, batch consistency and release. Sialic acid and monosaccharide analysis is required by ICH Q6B guidelines for biopharmaceutical characterization. ### Instrument configuration for native *N*-linked oligosaccharide characterization by HPAE-PAD/MS ![Instrument configuration for native N-linked oligosaccharide characterization by HPAE-PAD/MS](https://learning.sepscience.com/hs-fs/hubfs/AppNote1_UG.png?width=100&name=AppNote1_UG.png)Provide application installation instructions for coupling HPAE-PAD analyses to the Thermo Scientific™ Q Exactive™ family of mass spectrometers. This technical note focuses on using this set up to profile N-linked glycans released from glycoproteins. [![Read >>](https://no-cache.hubspot.com/cta/default/2322077/124d8d37-c552-4e10-963e-7b9df2110d18.png)](https://cta-redirect.hubspot.com/cta/redirect/2322077/124d8d37-c552-4e10-963e-7b9df2110d18)       ### Preparation of peptide *N*-Glycosidase F digests for HPAE-PAD analysis ![Preparation of peptide N-Glycosidase F digests for HPAE-PAD analysis](https://learning.sepscience.com/hs-fs/hubfs/AppNote2_UG.png?width=100&name=AppNote2_UG.png)The work shown here describes glycoprotein deglycosylation with PNGase F, followed by an efficient sample pretreatment using desalting and detergent removal spin columns to remove high concentrations of salt and detergents from small-scale PNGase F digestions. The HPAE-PAD technique was used to separate and detect the oligosaccharides. [![Read >>](https://no-cache.hubspot.com/cta/default/2322077/2ed55e36-bf6e-4a8e-8f40-3a8dcda19314.png)](https://cta-redirect.hubspot.com/cta/redirect/2322077/2ed55e36-bf6e-4a8e-8f40-3a8dcda19314)     ### Label-free Profiling of O-linked Glycans     ### Vanquish UHPLC System - Detect the Analyte with Charged Aerosol Detection   ### \[eBook\] Methods & Applications in BioPharmaceutical Characterization ![Thermo_Biop_eBookCover_eDM](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/eBook/Thermo_Biop_eBookCover_eDM.png?width=120&name=Thermo_Biop_eBookCover_eDM.png)Read about... - Intact Protein Analysis - Monoclonal Subunit Mass Analysis - Peptide Mapping Analysis - Peptide Mapping - Labeled Glycan Analysis - Unlabeled Glycan Analysis - Aggregate Analysis - Charge Variant Analysis [![Read >>](https://no-cache.hubspot.com/cta/default/2322077/18bbdd63-2155-44ff-875b-1203f6eab768.png)](https://cta-redirect.hubspot.com/cta/redirect/2322077/18bbdd63-2155-44ff-875b-1203f6eab768)     ### Label-Free Analysis by UHPLC with Charged Aerosol Detection of Glycans Separated by Charge, Size, and Isomeric Structure ![AppNote3_UGLabel-Free Analysis by UHPLC with Charged Aerosol Detection of Glycans Separated by Charge, Size, and Isomeric Structure](https://learning.sepscience.com/hs-fs/hubfs/AppNote3_UG.png?width=100&name=AppNote3_UG.png)This application note describes direct detection of native glycans as an alternative to the common techniques for glycan analysis that rely on derivatization reactions to render glycans detectable. The lack of a detectable chromophore in native glycans is overcome by using HPLC with charged aerosol detection, a detector that can quantitatively measure any non-volatile compound. [![Read >>](https://no-cache.hubspot.com/cta/default/2322077/ef1f13e6-f3dc-497c-9c27-ae64161bf61b.png)](https://cta-redirect.hubspot.com/cta/redirect/2322077/ef1f13e6-f3dc-497c-9c27-ae64161bf61b)     ### HPAE-PAD *N*-linked oligosaccharide profiling of IgG ![AppNote4_UGHPAE-PAD N-linked oligosaccharide profiling of IgG](https://learning.sepscience.com/hs-fs/hubfs/Companies/Thermo%20Fisher%20Scientific/BioPMicrosite/AppNote4_UG.png?width=100&name=AppNote4_UG.png)The goal of this study was to develop an HPAE-PAD method to profile the major neutral and charged asparagine-linked (N-linked) oligosaccharides from IgGs, including monoclonal antibodies. [![Read >>](https://no-cache.hubspot.com/cta/default/2322077/e934c007-1f41-4dbc-bb0b-239ebb2a36c3.png)](https://cta-redirect.hubspot.com/cta/redirect/2322077/e934c007-1f41-4dbc-bb0b-239ebb2a36c3)     ### Evaluating Protein Glycosylation in Limited-Quantity Samples by HPAE-PAD ![Evaluating Protein Glycosylation in Limited-Quantity Samples by HPAE-PAD](https://learning.sepscience.com/hs-fs/hubfs/AppNote6_UG.png?width=100&name=AppNote6_UG.png)In this study, the glycosylation of human transferrin (as a model of PSA) and PSA are investigated by HPAE-PAD using <10 µg of protein. Two methods are utilized to evaluate protein glycosylation. One method is used to investigate the N-linked oligosaccharides and glycan sialylation. A second method evaluates the potential presence of O-linked glycans by monitoring monosaccharide composition. 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