Development of a Novel, Simple, Sensitive, Rapid and Cost-Effective Diagnostic Assay for the Detection of Mesothelioma

Abstract

Malignant pleural mesothelioma is a cancer that has an extremely poor prognosis and is challenging to diagnose and treat. When diagnosed in earlier stages of cancer there are more treatment options available, and prognosis is better. Screening patient samples for the biomarker mesothelin is able to predict mesothelioma occurrence, so could be used to reduce the diagnostic delay and improve patient prognosis. However, the current technology able to detect mesothelin is designed for high-throughput screening and is too expensive for single sample testing – which is often needed as mesothelioma is a low incidence disease. As a result, this project aimed to develop a simple, low-cost device able to detect mesothelin that was better suited for mesothelioma screening than the current technology used.
In order to determine the device requirements needed to meet this aim, biomedical scientists were consulted on their preferences to create an ideal test specification. It was concluded that an electrochemical sensor would be suitable to meet these requirements. Sensor sensitivity is highly dependent on the antibody selected for detection and extensive screening resulted in a horseradish peroxidase-based sandwich assay format, optimised into an electrochemical sensor. The sensor produced a limit of detection of 11.3 ng/mL with manual use, well below the commonly used serum positive cut-off of 72 ng/mL, and therefore suitable for screening of serum-level concentrations of mesothelin.
Hardware, firmware, and software were designed to automate the testing procedure of the electrochemical sensor to meet criteria within the test specification. The developed automated device demonstrated a limit of detection of 104 ng/mL which allows for detection of mesothelin within pleural fluid that has a positive cut-off of 360ng/ml. Minor changes to the circuitry and fluid delivery system, should allow for automated detection of mesothelin within serum.
The research described in this thesis makes a significant and novel contribution towards the development of a simple, low cost, automated device capable of detecting mesothelin in clinical samples for early diagnosis of mesothelioma.

Date of Award	19 Mar 2022
Original language	English
Awarding Institution	University of Portsmouth
Supervisor	Marisa Van Der Merwe (Supervisor), Fiona Myers (Supervisor) & Sharon Glaysher (Supervisor)