A complete set of hyaluronan fragments obtained from hydrolysis catalyzed by hyaluronidase: Application to studies of hyaluronan mass distribution by simple HPLC devices
| Autor: | Corinne Loutelier-Bourhis, Didier Le Cerf, Jean-Claude Vincent, Marie-Noëlle Courel, Brigitte Deschrevel, Frédéric Tranchepain, Nicolas Levasseur |
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| Přispěvatelé: | Polymères, biopolymères, membranes (PBM), Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU), Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel), Spectrométrie de Masse Bio-organique [Rouen], Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2006 |
| Předmět: |
Bovine testicular hyaluronidase
Production of hyaluronan fragments Dispersity Size-exclusion chromatography Biophysics Hyaluronan hydrolysis Oligosaccharides Hyaluronoglucosaminidase 02 engineering and technology Mass spectrometry HPLC–SEC technique Biochemistry High-performance liquid chromatography Catalysis 03 medical and health sciences Hydrolysis chemistry.chemical_compound Hyaluronidase Glucosamine medicine Animals Hyaluronic Acid Molecular Biology Chromatography High Pressure Liquid 030304 developmental biology 0303 health sciences Molar mass Chromatography [CHIM.MATE]Chemical Sciences/Material chemistry Cell Biology 021001 nanoscience & nanotechnology Molecular Weight [CHIM.POLY]Chemical Sciences/Polymers chemistry Cattle 0210 nano-technology Reducing end assay medicine.drug |
| Zdroj: | Analytical Biochemistry Analytical Biochemistry, Elsevier Masson, 2006, 348 (2), pp.232-242. ⟨10.1016/j.ab.2005.09.042⟩ |
| ISSN: | 0003-2697 1096-0309 |
| DOI: | 10.1016/j.ab.2005.09.042 |
| Popis: | International audience; Hyaluronan (HA) has different biological functions according to its molar mass; short HA fragments are involved in inflammation processes and angiogenesis, whereas native HA is not. Physicochemically, studies of native HA hydrolysis catalyzed by bovine testicular hyaluronidase (HAase) have suggested that kinetic parameters depend on HA chain length. To study the influence of HA chain length in more detail, and to try to correlate the physicochemical and biological properties of HA, HA hydrolysis catalyzed by HAase was used in a new procedure to obtain HA fragments of different molar masses. HA fragments (10-mg scale) with a molar mass from 800 to 300,000 g mol−1 were prepared, purified using low-pressure size exclusion chromatography (SEC), lyophilized, and characterized in molar mass by either mass spectrometry or HPLC–SEC–multiangle laser light scattering. The polydispersity index of the purified fractions was less than 1.25. The complete set of HA standards obtained was used to calibrate our routine HPLC–SEC device using only a refractive index (RI) detector. We showed that the N-acetyl-d-glucosamine reducing end assay and the calibrated HPLC–SEC–RI gave equivalent kinetic data. In addition, the HPLC–SEC–RI furnished the mass distribution of the polysaccharide during its hydrolysis. |
| Databáze: | OpenAIRE |
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