Biopharmaceutical Glycosylation Analysis

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1 Biopharmaceutical Glycosylation Analysis

2 Glycosylation Analysis: Product Offering Molecular model of erythropoietin with complex N-linked glycans. Courtesy of M.R Wormald and R.A Dwek, Oxford Glycobioloy Institute and P.M. Rudd, NIBRT NIBRT s Biopharmaceutical Glycosylation Analysis provides: An automated high throughput glycan analysis platform. Fully detailed linkage and sequence analysis of glycans. Directly amenable for at-line PAT format as part of your QBD workflows. Technology is mobile and scalable which can be transferred directly to the Client s site. Competitive pricing structures.

3 Glycosylation Critical Quality Attributes (GCQAs) International Conference on Harmonisation (ICH) Harmonised Tripartite Guideline Q6B requires carbohydrate content, structure of carbohydrate chain and oligiosaccharides and glycosylation sites present on therapeutic proteins to be characterised as extensively as possible. The GCQAs required for biologics are listed below: All of the above GCQAs can be determined from NIBRT s analytical workflows. For details regarding the symbolic representation of glycans please refer to D.J. Harvey et al, PROTEOMICS, 9, (2009), 3796.

4 Analytical Complexity Tailored Analysis to Suit Your Needs Mass Spectrometry: HILIC profile of IgG N-glycans Initial Profiling: Sample preparation Glycan release Glycan labelling HILIC profiling Sialic acid linkage analysis Data interpretation and preliminary assignment. Structural Analysis: Exoglycosidase array digestion and profiling Anion exchange fractionation Data interpretation and preliminary assignment. Sialic Acid Speciation: Anion exchange analysis for sialic acid content Sialic acid speciation MALDI-Tof-MS ESI MS/MS analysis of individual glycans for Level 1-3 structural confirmation HILIC profile of N-glycans from Sigma Gonadatrophin Level 1 Level 2 Level 3 Level 4

5 Glycan Analysis Platform High-throughput glycan-analysis

6 GlycoBase v 3.0 GlycoBase v3.0 is a proprietary novel database that contains retention data (expressed as glucose unit values) for more than 400 2AB-labeled N-linked glycan structures by a combination of HILIC-fluorescence detection with exoglycosidase sequencing with confirmation via mass spectrometry. Each entry is comprehensively annotated and includes the following information: Screenshot from GlycoBase v3.0 A pictorial representation of the structure depicting monosaccharide sequence and linkages. HILIC-fluorescence retention time expressed as an average GU value, with standard deviation. Monosaccharide composition. Links to verified exoglycosidase digestion products. Related reference information A list of the subgroups in which the glycan can be found. NIBRT s proprietary versions of GlycoBase are updated on a regular basis and includes all published data and associated digestion profiles, together with confidential information generated by NIBRT. NIBRT offers a variety of deployment and customer support models to suit Client s specific requirements for access to GlycoBase v3.0.

7 Laboratory Core Competencies Liquid Chromatography: HILIC and ion exchange based glycosylation profiling. LC-MS of glycans. Modern suite of HPLC and UPLC instrumentation. Mass Spectrometry: MS characterisation of glycans using MALDI-Tof, ESI-MS, ESIMS/MS and ESI- MS n. Waters MALDI, Waters Q-Tof, Advion Nanomate and Agilent Ion Trap mass spectrometers. GlycoBioInformatics: GlycoBase a web based resource for HILIC data annotation. AutoGU an automated experimental data annotation tool. GlycoExtractor a tool for high throughput data extraction and searching. Electrophoretic Techniques: CE-LIF analysis of glycans. 1D and 2D SDS-PAGE. Proteomics and glycoproteomics

8 Further Information NIBRT s existing partners include: For further information about our Biopharmaceutical Glycosylation Analysis please contact us: Web: Phone:

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