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Multifunctional chromatography supports

Liddy, Alison Mary (2010)
Ph.D. thesis, University of Birmingham.

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Abstract

The aim of this study was to create a bi-layered packed bed chromatography support for the purification of nano-sized bioproducts. The effect of three different chemistry approaches, different solvent conditions, and microwave heating were investigated on construction of a bi-layered support. Sepharose CL-6B was activated with allyl glycidyl ether (routes 1&2). The inert outer layer was created in route 1 by reacting bromine with the allyl groups at the surface, followed by the addition of sodium hydroxide. Creation of the outer layer in route 2 was achieved by oxidation of surface groups with potassium permanganate. In both synthesises the allyl groups remaining were reacted with bromine and a charged amine ligand was coupled to the inner core. The activation step of route 3 resulted in the introduction of three membered epoxide groups throughout the support. Surface groups were reacted with sodium hydroxide or hydrochloric acid. Finally a charged amine ligand was coupled to the support by reacting trimethylamine hydrochloride with the remaining epoxide groups. Supports created by route 1 eliminated 91% of plasmid DNA binding whilst maintaining a high protein binding capacity. This was achieved by using DMS0 as the solvent in the bromination step and employing microwave heating. Route 2 proved to be the least successful in creating a bi-layered support. The beads created under hydrochloric acid-methanol conditions (route 3) reduced 91% of the plasmid DNA binding whilst maintaining a high protein binding capacity. This study revealed that microwave heating was a useful tool in the synthesis of chromatography supports. Subsequently, a comprehensive study was untaken investigating the effects of microwaves on numerous chromatography matrices.

Type of Work:Ph.D. thesis.
Supervisor(s):Thomas, Owen and Theodossiou, Eirini
School/Faculty:Colleges (2008 onwards) > College of Engineering & Physical Sciences
Department:School of Chemical Engineering
Subjects:TP Chemical technology
QD Chemistry
Institution:University of Birmingham
ID Code:594
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
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