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Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing / KAROL SZUBA-JABLONSKI

Swansea University Author: KAROL SZUBA-JABLONSKI

DOI (Published version): 10.23889/SUthesis.65875

Abstract

Ultrabithorax (Ubx) is a Hox transcription factor, which self-assembles into protein films and fibres at the air-water interface. Ubx materials are bio- and cyto-compatible, and can be functionalised with DNA, proteins, and growth factors to benefit from functions such as supporting cell proliferati...

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Published: Swansea University, Wales, UK. 2024
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
Supervisor: Meissner, K. ; Dunstan, P.
URI: https://cronfa.swan.ac.uk/Record/cronfa65875
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spelling v2 65875 2024-03-20 Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing e07eedbd90124d01ef28aa35059bfac2 KAROL SZUBA-JABLONSKI KAROL SZUBA-JABLONSKI true false 2024-03-20 Ultrabithorax (Ubx) is a Hox transcription factor, which self-assembles into protein films and fibres at the air-water interface. Ubx materials are bio- and cyto-compatible, and can be functionalised with DNA, proteins, and growth factors to benefit from functions such as supporting cell proliferation. This thesis investigated the optical, electrical, and mechanical properties of Ubx fusion fibres in conditions relevant to applications in biosensing and tissue engineering. The steady-state and time-resolved spectra of the fibres were measured using an ultrafast laser source. The dityrosine emission peak red-shifted by 50nm in fibres compared to Ubx fusion solutions, and resonance energy transfer between dityrosine and enhanced green fluorescent protein (EGFP) was observed in the EGFP-Ubxprotein fusion. The electrical properties of Ubx fusion fibres were tested, and anincrease in their electrical conductivity by 3 orders of magnitude was measuredwith rising relative humidity. Tensile tests of Ubx fusion fibres revealed that thefibres were less extensible but stronger after prolonged storage and rehydration,which could influence the design of Ubx materials for tissue engineering. Significant binding of DNA aptamers to Ubx in solution and in fibres was observed. A fluorescence-based method to assess binding eciency of bacteria and the SARSCoV-2 spike RBD protein was developed. Some indication of increased specific pathogen binding was observed using long oligonucleotide sequences bound to Ubx fusion fibres, and improvements to this system were suggested. Alginate and collagen are biomaterials widely used in tissue engineering. Preliminary experiments were conducted that showed the feasibility of production of composite materials made of Ubx and alginate or collagen, which could be applied to createfunctional tissue scaffolds. Ubx-based platforms and composite materials couldbenefit from the intrinsic properties of Ubx, and Ubx functionalisation with proteins,growth factors and DNA aptamers for applications in biosensing and tissueengineering. E-Thesis Swansea University, Wales, UK. Ubx, protein-based materials, biosensing, optical properties, electrical properties, mechanical properties, aptamers, composite biomaterials, fluorescence, tensile testing, pathogen capture 4 1 2024 2024-01-04 10.23889/SUthesis.65875 Part of this thesis has been redacted to protect personal information. COLLEGE NANME COLLEGE CODE Swansea University Meissner, K. ; Dunstan, P. Doctoral Ph.D Swansea University/SURES Not Required Swansea University/SURES 2024-04-28T17:34:37.2567247 2024-03-20T15:22:18.1777018 Faculty of Science and Engineering School of Biosciences, Geography and Physics - Physics KAROL SZUBA-JABLONSKI 1 65875__29765__826c48c6551c4b9db562b1930b16ae8e.pdf 2023_Szuba-Jablonski_K_final.65875.pdf 2024-03-20T15:39:50.4091120 Output 66572720 application/pdf E-Thesis true Copyright: The author, Karol Szuba-Jablonski, 2024. true eng
title Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
spellingShingle Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
KAROL SZUBA-JABLONSKI
title_short Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
title_full Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
title_fullStr Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
title_full_unstemmed Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
title_sort Characterisation and Functionalisation of Ultrabithorax Materials for Biosensing
author_id_str_mv e07eedbd90124d01ef28aa35059bfac2
author_id_fullname_str_mv e07eedbd90124d01ef28aa35059bfac2_***_KAROL SZUBA-JABLONSKI
author KAROL SZUBA-JABLONSKI
author2 KAROL SZUBA-JABLONSKI
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publishDate 2024
institution Swansea University
doi_str_mv 10.23889/SUthesis.65875
college_str Faculty of Science and Engineering
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hierarchy_top_id facultyofscienceandengineering
hierarchy_top_title Faculty of Science and Engineering
hierarchy_parent_id facultyofscienceandengineering
hierarchy_parent_title Faculty of Science and Engineering
department_str School of Biosciences, Geography and Physics - Physics{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Biosciences, Geography and Physics - Physics
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description Ultrabithorax (Ubx) is a Hox transcription factor, which self-assembles into protein films and fibres at the air-water interface. Ubx materials are bio- and cyto-compatible, and can be functionalised with DNA, proteins, and growth factors to benefit from functions such as supporting cell proliferation. This thesis investigated the optical, electrical, and mechanical properties of Ubx fusion fibres in conditions relevant to applications in biosensing and tissue engineering. The steady-state and time-resolved spectra of the fibres were measured using an ultrafast laser source. The dityrosine emission peak red-shifted by 50nm in fibres compared to Ubx fusion solutions, and resonance energy transfer between dityrosine and enhanced green fluorescent protein (EGFP) was observed in the EGFP-Ubxprotein fusion. The electrical properties of Ubx fusion fibres were tested, and anincrease in their electrical conductivity by 3 orders of magnitude was measuredwith rising relative humidity. Tensile tests of Ubx fusion fibres revealed that thefibres were less extensible but stronger after prolonged storage and rehydration,which could influence the design of Ubx materials for tissue engineering. Significant binding of DNA aptamers to Ubx in solution and in fibres was observed. A fluorescence-based method to assess binding eciency of bacteria and the SARSCoV-2 spike RBD protein was developed. Some indication of increased specific pathogen binding was observed using long oligonucleotide sequences bound to Ubx fusion fibres, and improvements to this system were suggested. Alginate and collagen are biomaterials widely used in tissue engineering. Preliminary experiments were conducted that showed the feasibility of production of composite materials made of Ubx and alginate or collagen, which could be applied to createfunctional tissue scaffolds. Ubx-based platforms and composite materials couldbenefit from the intrinsic properties of Ubx, and Ubx functionalisation with proteins,growth factors and DNA aptamers for applications in biosensing and tissueengineering.
published_date 2024-01-04T17:34:35Z
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