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    Development of a Non-Invasive Proteomic Approach to Profiling Molecular Changes in the Microenvironment to Investigate Stages of Breast Health

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    MPhil (4.268Mb)
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    Publication date
    2020
    Author
    George, Amy L.
    Supervisor
    Sutton, Chris W.
    Poterlowicz, Krzysztof
    Keyword
    Breast cancer
    Biomarkers
    Proteomics
    Nipple aspirate fluid
    Liquid biopsy
    Non-invasive
    Molecular changes
    Microenvironment
    Rights
    Creative Commons License
    The University of Bradford theses are licenced under a Creative Commons Licence.
    Institution
    University of Bradford
    Department
    Faculty of Life Sciences
    Awarded
    2020
    
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    Abstract
    Early detection of breast cancer is critical for increasing survival rates. However, currently available screening strategies provide ambiguous results, leaving invasive tissue biopsy procedures necessary for definitive diagnosis. Considerable efforts have investigated using nipple aspirate fluid (NAF), a liquid biopsy rich in proteins representative of the breast microenvironment, as a non-invasive source of early detection biomarkers. However, by using traditional two-dimensional discovery proteomic approaches, many technical challenges of using NAF have limited analysis of large sample sizing: such as low expressed volume (<10µL) or insufficient analytical material (<200µg protein). Following non-invasive collection by manual massage, we developed a one-dimensional sample preparation workflow that reduced sample handling steps, minimised sample losses and increased sample throughput to 96-samples by using a PVDF-membrane based system, which was ideally suited to the NAF proteome. Samples were prepared within a single working day, and results correlated significantly with conventional in-solution protocols. ​Proteins typically associated with the dysregulation of innate immune response and haemostatic pathways had a significantly altered proteome profile in response to breast cancer. Overall, our new workflow will allow future studies to take a more high-throughput approach, revealing biomarkers for breast cancer early detection, and providing a real impact.
    URI
    http://hdl.handle.net/10454/19058
    Type
    Thesis
    Qualification name
    MPhil
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