Separation of blood group A-active oligosaccharides by high-pressure liquid affinity chromatography using a monoclonal antibody bound to concanavalin A silica |
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| Authors: | J Dakour A Lundblad D Zopf |
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| Affiliation: | 1. Department of Clinical Chemistry, University Hospital, S-221 85, Lund, Sweden;2. National Cancer Institute, Building 10 Room 2N102, National Institutes of Health, Bethesda, Maryland 20892 USA;1. Department of Research Core Services, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44106, USA;2. Department of Nutrition, Case Western Reserve University, Cleveland, OH 44106, USA;3. Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, OH 44272, USA;4. Department of Quantitative Health Sciences, Cleveland Clinic, Cleveland, OH 44195, USA;5. Merck Research Laboratories, 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA;6. Department of Hepatology and Gastroenterology, Digestive Disease Institute, Cleveland Clinic, Cleveland, OH 44195, USA;1. Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, The Salgrenska Academy, University of Gothenburg, Mölndal, Sweden;2. Department of Pathology & Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA;3. Waters Corporation, Milford, MA, USA;4. Chromatographic Sciences Department, PPD Laboratories, Richmond, VA, USA;5. Medical & Scientific Relations Division, Alzheimer''s Association, Chicago, IL, USA;6. Institute for Reference Materials and Measurements (IRMM), Joint Research Centre, European Commission, Geel, Belgium;7. UCL Institute of Neurology, London, UK;1. Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences (C4N), Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium;2. Department of Analytical Chemistry and Pharmaceutical Technology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium;3. Waters Corporation, Connexion Business Park, Brusselsesteenweg 500, 1731 Zellik, Belgium;4. Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757, USA;1. Department of Chemical and Biochemical Engineering, University of Western Ontario, London, Canada;2. Faculty of Engineering & Applied Science, Memorial University of Newfoundland, St John''s, Canada;1. Michael Barber Centre for Mass Spectrometry, Manchester Institute of Biotechnology, University of Manchester, Princess Road, Manchester M1 7DN, UK;2. ChELSI Institute, Department of Chemical and Biological Engineering, University of Sheffield, Mappin Street, Sheffield S1 3JD, UK;3. Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK;4. Manchester Pharmacy School, University of Manchester, Stopford Building, Oxford Road, Manchester M13 9PT, UK |
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| Abstract: | A column for high-pressure liquid affinity chromatography is prepared by binding a murine monoclonal anti-blood group A antibody of IgM isotype to concanavalin A-coated silica particles. The column specifically retards blood group A-active oligosaccharides with the nonreducing immunodominant trisaccharide sequence, GalNAc alpha 1-3(Fuc alpha 1-2)Gal beta 1- ..., and separates three A-active oligosaccharides with different core structures. Retention of the oligosaccharides on the column diminishes with increasing temperatures, permitting thermal elution in the range 25-50 degrees C. |
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