Droplet interface bilayers for drug discovery
File(s)
Author(s)
Strutt, Robert
Type
Thesis or dissertation
Abstract
Droplet interface bilayers (DIB) formed at the contact of two aqueous in oil emulsion droplets, show huge potential in a range of fields. This work is concerned with the application of DIB technology in drug discovery and industrially relevant processes as a model system for measuring passive, biological membrane diffusion. A current limitation of the field has been an over reliance on fluorescence as a read out for permeant concentration. This thesis presents two separate methods for label-free DIB translocation experiments engineered with a suite of fabrication techniques, in situ data analysis pipelines and semi automation to perform UV spectroscopy in nL volume droplets. In one method, one of the DIB droplets is quantitatively measured and in the other, full translocation dynamics are obtained through mobilisation of a beam path, across multiple DIB droplet pairs. These methods unlock highly relevant permeant structure - permeability relationships decoupled in a DIB environment.
With these developed methods, studies were performed across a series of distinct chemical and biophysical spaces. Systematic functional group changes afforded a correlation between DIB permeability and drug-likeness. Within a biological dataset it was found that native compounds behaved in a biomimetic fashion within DIBs, highlighting the role of compound labelling. Over 30 compounds were assayed across wide chemical space, broadly representative of drug design with a MW range of 150 to 1130 Da. Permeant structure and physicochemical metrics were linked to the DIB permeability. It was found that oil partition as a model for bilayer permeability is inherently flawed, opening DIBs as a replacement of current, industrially applied technologies. Finally, efforts were placed on a more mature synthetic membrane. A bacterial mechanosensitive transmembrane protein was gated in a concentration dependent manner through lipid composition of the membrane...
With these developed methods, studies were performed across a series of distinct chemical and biophysical spaces. Systematic functional group changes afforded a correlation between DIB permeability and drug-likeness. Within a biological dataset it was found that native compounds behaved in a biomimetic fashion within DIBs, highlighting the role of compound labelling. Over 30 compounds were assayed across wide chemical space, broadly representative of drug design with a MW range of 150 to 1130 Da. Permeant structure and physicochemical metrics were linked to the DIB permeability. It was found that oil partition as a model for bilayer permeability is inherently flawed, opening DIBs as a replacement of current, industrially applied technologies. Finally, efforts were placed on a more mature synthetic membrane. A bacterial mechanosensitive transmembrane protein was gated in a concentration dependent manner through lipid composition of the membrane...
Version
Open Access
Date Issued
2022-03-21
Date Awarded
2022-05-01
Copyright Statement
Creative Commons Attribution NonCommercial Licence
License URL
Advisor
Ces, Oscar
Brooks, Nicholas
Gould, Ian
Publisher Department
Chemistry
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)