Development and Translation of Mass Spectrometry in the Fields of Non-Adherence and Covid-19

Non-adherence is the failure to follow prescribed treatment regimens. In cardiometabolic diseases (e.g., hypertension, type 2 diabetes mellitus etc), non-adherence is directly linked with increased risk of hospitalisation and early all-cause mortality. These endpoints are completely avoidable. However, the incidence of non-adherence may occur in >25% of patients. New chemical adherence tests (CAT) have recently been adopted for clinical use in the UK (circa. 2014), and these represent the gold-standard measure for determining adherence. Using liquid-chromatography tandem mass spectrometry (LC-MS/MS), biomatrices (e.g., urine/plasma) are tested qualitatively for the presence of medications to indicate ingestion. Clinically, CAT is not widespread (only available in specialist clinics), it is slow (>60 minutes per sample), and it poses a risk of false results (quantitative data may identify sporadic dosing). This thesis developed a set of guidelines regarding the use of CAT, which was published and endorsed by the European Society of Hypertension Adherence Working Group. Then, Fast (<6.5 minute) and UltraFast (<45 second) LC-MS/MS quantitative assays were developed, optimised, and validated to provide platforms amendable for high-throughput testing. An automated R-based validation processor was also built to deal with untenable validation datasets during LC-MS/MS development.

The initial aims of the thesis then shifted due to the COVID-19 pandemic. Here, the Department of Health and Social Care (DHSC) set a UK-wide directive to rapidly develop and implement novel SARS-CoV-2 testing devices for mass screening (“Operation Moonshot”). The latter half of this work follows the role of this thesis in Operation Moonshot. A proteomic LC-MS/MS assay was developed. Using a novel viral peptide-capture approach, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid peptides were successfully resolved from human saliva for the determination of active infection. This assay was then tested against the gold-standard, reverse transcriptase polymerase chain reaction (RT-PCR), to assess its clinical utility.