Advanced tandem quadrupole mass spectrometer with an automated sample extractor for a multiuser interdisciplinary translational research facility

There have been billions of pounds spend in research on new tests for diseases (biomarkers) but only a handful have made it to clinical use. The reasons are that there is often a lack of direct clinical question asked by research labs studying biomarkers and often a lack of robust method development and validation such that it can be used clinically. The van Geest Multi-OMIC facility is a world leader in the discovery and translation of biomarkers. We, in joint partnership with the University Hospitals of Leicester have developed the National Centre of Adherence Testing (NCAT), for assessing whether patients are adhering to their prescribed medications. This work has had impact not just on patient outcomes and international guidelines but to the design of clinical trials. The van Geest Multi-OMIC facility participated in the DHSC Moonshot Project. This national programme established a framework for rapid development, validation and implementation of a triple quadrupole mass spectrometer (LC-MS) method for the SARS-CoV-2 virus in swabs. The programme took about 5 months to develop a valid method for use clinically. The LC-MS instrument is state of the art and was provided with minimal fee for a year by the manufacturer Waters and is on site in our laboratory.
LC-MS have exquisite levels of sensitivity and incredible levels of ability to identify only the protein of interest (specificity). An example of this is the use of LC-MS in identification of doping by elite athletes. Before samples can be loaded on the LC-MS they need to be prepared. These manual steps of purifying and isolating are slow and can lead to errors. Therefore, in this proposal we are requesting also an Agilent Bravo liquid handling unit to enable us to do a wide range of sample preparation approaches in a swift, reproducible and robust manner.
We request the purchase of the above LC-MS to develop four main clinical projects that can be translated into clinically relevant assays using lessons learnt from the Moonshot programme and NCAT. The equipment offers very good value for money as it is being offered at a 53.5% discount by Waters.
The first project is to develop a method that can detect multiple viruses seen in the winter such as influenza and Covid-19- in a rapid and scalable manner. We can also use MS to identify variants of viruses and look to see whether this can be used in determining viral persistence in Long COVID.
The second project will deliver vastly improved methods for NCAT. It will reduce analysis time from 80 minutes to between 45 seconds to 3.5 minutes. Currently, due to the time taken, the method is available mainly for hypertension clinics. The new methods will enable the use to be expanded to other conditions including heart failure and rheumatoid arthritis, in hospitalised patients and those seeing their GP. We will also develop capabilities to study large research populations and the tailoring of medications according to an individual's medication blood level.
The third project will lead a pioneering effort to develop ten methods to study a key pathway in blood pressure regulation- the renin-angiotensin aldosterone system (RAAS). These methods will help understand subgroups of hypertension and response to medications.
The fourth project will allow remote patient monitoring of patients with long term conditions such as diabetes, thyroid disease and heart disease. Patients will self-collect their blood by finger prick, and we will develop methods for measurements of routine tests such their liver and kidney function. Successful completion will help provide robustness in patient management remotely- especially during future pandemics and free resources both in patient time and of the NHS.

These projects are exemplars of the type of project that will enable us to swiftly validate and gain accreditation for assays that can then be taken straight into NHS laboratories.

The Waters Triple quadrupole XS (TQ-XS) is a market leading mass spectrometer which demonstrates exquisite levels of sensitivity, robustness and specificity. It contains a modified detector which enables six orders of magnitude of linear dynamic range. Similar to many TQ mass spectrometers we are able to develop multiplex assays ( between 10-100 analytes per assay). However, with the Waters UPLC separation unit, we are able to ensure robust consistency in separation performance which allows assays to take advantage of scheduled SRM windows which maximises the duty cycle of the instrument. The instrument has been on the market for 5 years and has thus proven track record in a range of complex analyses with high performance. One consequence of the performance of the LC-MS system is that run times can be significantly reduced increasing throughput of samples considerably. On a 3min run time, ~480 samples a day can be analysed. Thus, there is a need to automate the sample preparation so that the labour intensive sample preparation steps can be automated into 96 well plate format. This will greatly enhance our ability to take advantage of the analytical performance of the LC-MS. The Agilent Assaymap Bravo is a liquid handling/sample preparation unit that is widely used in Omics laboratories across the globe. It will allow larger numbers of samples to be prepared in shorter amount of time with greater consistency. It was designed specifically for upstream of LC-MS sample preparation techniques and allows automated immunoaffinity capture, digestion and desalting steps. Liquid/liquid extraction (LLE) and solid phase extraction(SPE) can also be done on the same platform. It can also be used to prepare samples for protein quantitation. The projects detailed in the CfS will benefit from access to high throughput technology and will help build a infrastructure that can be pivoted at any stage towards future pandemics as well as providing a model of clinical assay development.