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Amélioration instrumentale de la chromatographie de partage centrifuge en vue de la purification de molécules très polaires

Abstract : The aim of this thesis is to develop a new centrifugal partition chromatography (CPC) instrument in order to purify highly polar molecules. CPC is a preparative technique for the separation of molecules using a solvent system composed of two immiscible liquids. This manuscript describes the different protein purification techniques used in industrial production process. A focus is made on the use of aqueous biphasic systems for the purification of biomolecules, which represents a real trend in the industry thanks to its low cost, scaling simplicity and especially the favorable environment that it provides to biomolecules. Thus, based on the advantages of these solvent systems known as Aqueous Two Phase Systems (ATPS), CPC could provide additional performances to purify proteins at lower cost. To respond to this industrial challenge, it is necessary to develop both innovative chromatographic methods and new devoted instruments. Indeed, current CPC instruments are not compatible with Good Manufacturing Practices due to the presence of Teflon seals which prevents the possibility of sterilizing the instruments. The manufacture of a new monobloc instrument entirely made of titanium was achieved thanks to the 3D printing technology. The purpose of this thesis is the evaluation of this new column performance in order to determine its applicability to biomolecules purification. A special attention is also provided to the injection of very small sample volumes in order to facilitate method development
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Submitted on : Thursday, May 3, 2018 - 4:29:06 PM
Last modification on : Friday, April 17, 2020 - 10:46:09 AM
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  • HAL Id : tel-01784638, version 1



Feriel Bouiche. Amélioration instrumentale de la chromatographie de partage centrifuge en vue de la purification de molécules très polaires. Chimie analytique. Université de Lyon, 2018. Français. ⟨NNT : 2018LYSE1003⟩. ⟨tel-01784638⟩



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