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Title: Development of high-throughput technologies for the study of drug-membrane interactions
Author: Stanley, Claire Elizabeth
ISNI:       0000 0004 2706 1890
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2011
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Understanding how drug molecules interact with our body and what effects they may induce as a result is of fundamental concern to the pharmaceutical industry. Crucially we want to use this knowledge to our advantage during the drug discovery process in order to manipulate a drug’s efficacy in vivo - therefore the development of new technologies, able to effectively screen numerous desirable drug-membrane interactions, is of key importance. The first half of this thesis details the development of a vesicle leakage assay, as a means to assess the effect of cationic amphiphilic drugs (CADs) on model lipid membranes. Having demonstrated the reproducibility of the assay, the assay was transferred into a microfluidic format where water-in-oil droplet systems act as individual experimental vessels. As such, it has been demonstrated that the use of fluorescence lifetime techniques can provide a way in which to translate this assay into a high-throughput format. The second part of the thesis is concerned with using droplet interface bilayers (DIBs) as a means to probe the effect of exogenous species upon a single lipid bilayer, as opposed to bulk vesicle populations. Several advantages exist for using such a system, including for example the ease with which one is able to control the composition of the aqueous compartments on either side of the bilayer and to form asymmetric bilayers. An assay, involving the use of pH gradients, is detailed, where proof-of-concept experiments illustrate that pH sensitive dyes could be used to report the extent to which lipid bilayers are perturbed by drug molecules for example. Furthermore, a novel automated approach has been developed, offering advantages over the manual manipulation of lipid-containing droplets for DIB formation, where a microfluidic approach is used for their generation in high-throughput. Consequently, this approach enables the formation of multiple DIBs, where the composition may be differed and the droplet dimensions controlled, enabling the formation of DIB networks that can be arranged in either two- or three-dimensions.
Supervisor: Ces, Oscar ; de Mello, Andrew ; Gee, Tony Sponsor: EPSRC ; GSK ; Society of the Chemical Industry
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral