Chromatographic fractionation of yeast extract: A strategy to identify physicochemical properties of compounds promoting CHO cell culture
| Autor: | Romain Kapel, Laurent-Michel Bonanno, Ivan Marc, Eric Olmos, Isabelle Chevalot, Arnaud Aymes, Mathilde Mosser, Annie Marc |
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| Přispěvatelé: | Laboratoire Réactions et Génie des Procédés (LRGP), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Bio-Springer, Immuno-Endocrinologie Cellulaire et Moléculaire [Nantes] (IECM), Université de Nantes (UN)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de la Recherche Agronomique (INRA), Laboratoire des Sciences du Génie Chimique (LSGC), Institut National Polytechnique de Lorraine (INPL)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Lorraine (UL), Institut National de la Recherche Agronomique (INRA)-Université de Nantes (UN)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS) |
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
0106 biological sciences
Chromatographic fractionation Size-exclusion chromatography Bioengineering Fractionation 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Hydrophobic effect 03 medical and health sciences [SPI]Engineering Sciences [physics] Size exclusion chromatography 010608 biotechnology Yeast extract [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering ComputingMilieux_MISCELLANEOUS Hydrophobic interaction chromatography 030304 developmental biology 0303 health sciences Chromatography Chemistry Cell growth Hydrophilic interaction chromatography Anion exchange chromatography CHO cell growth Separation process Chemically defined medium |
| Zdroj: | Process Biochemistry Process Biochemistry, Elsevier, 2012, 47 (7), pp.1178-1185. ⟨10.1016/j.procbio.2012.04.015⟩ |
| ISSN: | 1359-5113 0032-9592 |
| DOI: | 10.1016/j.procbio.2012.04.015⟩ |
| Popis: | International audience; The study proposes to get a better knowledge of the physicochemical properties of yeast extract (YE) molecules involved in the improvement of CHO cell growth and to reduce YE complexity without losing positive effects. Various chromatographic processes were implemented for fractionation of a nanofiltrated YE (nYE). The nYE was first fractionated by one-step preparative chromatography, either with anion exchange (AEC), hydrophobic interaction (HIC) or size exclusion (SEC) methods. After analysis of its main components, each fraction was added in a control chemically defined medium to assess its impact on CHO cell growth. Results mainly underlined that AEC was the most selective separation process to purify nYE in one step without decreasing cell growth promoting effect. A three-step chromatographic process including successive AEC. HIC, and SEC was then developed to refine the physicochemical properties of nYE compounds. Among fractions that triggered similar cell growth promoting effect than nYE, one also improved IgG specific production. It mainly included cationic and hydrophilic peptides with a great proportion of lysine and arginine, low quantities of polysaccharides and no nucleic acids. Thus, this study allowed us to deepen the YE contribution to animal cell culture as well as to evaluate fractionation strategies to simplify such a complex mixture. |
| Databáze: | OpenAIRE |
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