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Hydrogel encapsulated droplet interface bilayer networks as a chassis for artificial cells and a platform for membrane studies

Baxani Kamal, Divesh 2017. Hydrogel encapsulated droplet interface bilayer networks as a chassis for artificial cells and a platform for membrane studies. PhD Thesis, Cardiff University.
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Abstract

There has been increasing interest in droplet interface bilayers (DIBs) as novel devices for the study of lipid membranes and the development of artificial cell systems. Although DIBs have demonstrated to be useful in a number of laboratory applications, their wider use is hampered by a limited ability to exist untethered and remain mechanically stable beyond controlled laboratory environments. In this thesis, a microfluidic system is developed which enables the facile generation of hydrogel-encapsulated DIB networks which are freestanding and can exist in air, water and oil environments, without compromise to their ability to interface with the surrounding environment. Electrophysiology is employed in order to demonstrate the formation of bilayers between the encapsulated DIBs (eDIBs) and their external environment, achieved via the incorporation of the transmembrane pore α-Hemolysin. The eDIBs produced here are able to form higher-order structures akin to tissues via their assembly and adherence to one another, further demonstrating their potential to act as a chassis for artificial cells. Furthermore, the potential of eDIBs to be used as a platform for membrane studies is demonstrated via their use as a high-throughput array for membrane disruption fluorescence measurements using a plate reader, which makes use of the ability of eDIBs to be generated in large numbers as well as to be mechanically handled and placed in the wells of a 96-well plate. Fluorescence measurements were taken on up to 47 eDIBs simultaneously, and were able to detect bilayer leakage through pores as well as bilayer failure. The above experiments comprise the design, manufacture and use of a novel kind of DIB construct as a chassis for artificial cells and a platform for high-throughput membrane studies. It is proposed that eDIBs may help in realising the unfulfilled potential of DIB networks in applications in healthcare and beyond.

Item Type: Thesis (PhD)
Date Type: Completion
Status: Unpublished
Schools: Pharmacy
Subjects: R Medicine > R Medicine (General)
R Medicine > RM Therapeutics. Pharmacology
Date of First Compliant Deposit: 25 June 2018
Last Modified: 05 Jul 2019 02:05
URI: http://orca-mwe.cf.ac.uk/id/eprint/112707

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