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Title: Chemical and biological approaches to enhance the bacteriophage as a probe in molecular recognition
Author: Lobo, Daniela P.
ISNI:       0000 0004 7431 6796
Awarding Body: University of Warwick
Current Institution: University of Warwick
Date of Award: 2017
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The aim of the work presented in this thesis was to explore the structural and molecular features of a filamentous bacteriophage for incorporation in biosensing platforms. To tune the bacteriophage surface as a scaffold for molecular recognition, chemical and biological protocols aimed at controlling and enhancing its intrinsic properties were developed. A key part of the work was aimed at anchoring bacteriophage particles by one-end to surfaces, and on labelling its viral capsid with dyes, antibodies and DNA molecules. The tolerance of the bacteriophage for modifications, as well its simplicity and robustness, makes it an attractive probe for biosensors. Furthermore, the filamentous structure of the bacteriophage was demonstrated to be a valuable feature in both microscopy- and spectroscopy-based biosensors. The ability of the bacteriophage to align under flow was considered as a simple, yet efficient approach for the detection of wall shear stress and pathogenic bacteria. A novel method to detect wall shear stress (WSS) was developed by using a fluorescently decorated bacteriophage particle anchored by one-end to a surface. The response of this filamentous nanosensor to flow variations was tracked under a microscope, and gave valuable information on the shear flow of a fluid passing over a surface. Using a custom-made analysis tool and an algorithm, we were able to derive the wall shear stress on the point of attachment of the nanosensor, using endothelial cells as a model system. The proof-of-concept to this work highlighted how a simple bacteriophage construct can be use as a nanosensor for imaging and mapping flow. Linear dichroism (LD) spectroscopy also explores the high aspect ratio of the filamentous bacteriophage. Linear Diagnostics Ltd., a startup company in Birmingham, exploits the natural LD properties of the bacteriophage and integrated it with a biosensing platform targeting pathogenic bacteria. The work reported in this thesis mainly focused on developing alternative approaches that could improve the biosensor sensitivity and simplicity, by modifying the bacteriophage scaffold, further demonstrating its versatility.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: QD Chemistry