Comparison of methods for the analysis of therapeutic immunoglobulin G Fc-glycosylation profiles—Part 1: Separation-based methods
| Autor: | Maria Maier, Markus Haberger, Samnang Tep, Bernd Maier, Dietmar Reusch, Manfred Wuhrer, Zoltan Szabo, Patrick Bulau, Jo Wegstein, Ronny Kloseck, Boris Zimmermann, Michaela Hook, Nadja Alt |
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| Rok vydání: | 2015 |
| Předmět: |
Glycosylation
CHO CE-LIF 2-AB 01 natural sciences High-performance liquid chromatography Mass Spectrometry MALDI-MS Immunoglobulin G immunoglobulin G chemistry.chemical_compound Cricetinae ionization-mass spectrometry HR Immunology and Allergy capillary electrophoresis-laser induced fluorescence HPAEC-PAD Chromatography High Pressure Liquid high-throughput 0303 health sciences Fc biology HPAEC Chemistry IAB matrix-assisted laser desorption Immunoglobulin Fc Fragments Antibodies Monoclonal 8-aminonaphthalene-1 glycan analysis DNA-sequencer-aided fluorophore-assisted carbohydrate electrophoresis 3. Good health method comparison 2-aminobenzamide 6-trisulfonic acid electrospray ionization-mass spectrometry CCGE monoclonal antibody (mAb) Antibody HILIC-UPLC Glycan high performance liquid chromatography IgG IgG glycosylation ANTS Immunology APTS CHO Cells 8-aminopyrene-1 fragment APTS labeling 03 medical and health sciences Cricetulus Capillary electrophoresis hydrophilic interaction liquid chromatography-ultra high performance liquid chromatography 6-trisulfonate InstantAB labeling Animals Fab Bioprocess 2-AB labeling antigen-binding cartridge-based capillary gel electrophoresis 030304 developmental biology mAb Chromatography high-performance anion exchange chromatography with pulsed amperometric detection high resolution 010401 analytical chemistry ESI-MS DNA analyzer DSA-FACE Chinese hamster ovary 0104 chemical sciences carbohydrates (lipids) monoclonal antibody fragment crystallizable biology.protein HPLC Reports |
| Zdroj: | mAbs, 7(1), 167-179 mAbs |
| ISSN: | 1942-0870 1942-0862 |
| DOI: | 10.4161/19420862.2014.986000 |
| Popis: | Immunoglobulin G (IgG) crystallizable fragment (Fc) glycosylation is crucial for antibody effector functions, such as antibody-dependent cell-mediated cytotoxicity, and for their pharmacokinetic and pharmacodynamics behavior. To monitor the Fc-glycosylation in bioprocess development, as well as product characterization and release analytics, reliable techniques for glycosylation analysis are needed. A wide range of analytical methods has found its way into these applications. In this study, a comprehensive comparison was performed of separation-based methods for Fc-glycosylation profiling of an IgG biopharmaceutical. A therapeutic antibody reference material was analyzed 6-fold on 2 different days, and the methods were compared for precision, accuracy, throughput and other features; special emphasis was placed on the detection of sialic acid-containing glycans. Seven, non-mass spectrometric methods were compared; the methods utilized liquid chromatography-based separation of fluorescent-labeled glycans, capillary electrophoresis-based separation of fluorescent-labeled glycans, or high-performance anion exchange chromatography with pulsed amperometric detection. Hydrophilic interaction liquid chromatography-ultra high performance liquid chromatography of 2-aminobenzamide (2-AB)-labeled glycans was used as a reference method. All of the methods showed excellent precision and accuracy; some differences were observed, particularly with regard to the detection and quantitation of minor glycan species, such as sialylated glycans. |
| Databáze: | OpenAIRE |
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