Developing Drug Discovery Assays for Primary Human Immune Cells by High-Throughput MALDI-TOF Mass Spectrometry
MALDI-TOF mass spectrometry (MS) is a powerful MS method that has emerged as an exciting technology high-throughput drug discovery workflows. Publications from our lab and pharma industry are now demonstrating the development of MALDI-based in vitro assays that measure the products of enzymatic reactions. The development of cell-based assays by MALDI, particularly for primary immune cells such as T-cells and macrophages is, however, less well established. In this project, we will characterise and develop
assays for primary human immune cells playing a role in inflammatory disease and immunooncology by MALDI-TOF MS and scale those methods for compound screening workflows amenable to drug discovery.
MALDI-TOF mass spectrometry is a powerful tool which has long been used in the pharmaceutical industry for the analysis of proteins, peptides and other bioactive molecules. However, until recently the throughput of MALDI-TOF was insufficient, particularly compared to fluorescence-based methods, for high-throughput screening workflows. Recently, next generation MALDI-TOF instruments have emerged with improved spatial resolution and 10kHz lasers. Together these instruments enable up to a 20x increase in throughput versus legacy systems. The emergence of these systems, coupled with advances in nanolitre liquid handling technologies for sample preparation, provide the opportunity for applying MALDI-TOF platforms to high-throughput biological assays and screening workflows such as those typically used in industrial screening laboratories. Indeed, examples of such workflows have now been published (Heap et al, Analyst, 2019; Heap et al, SLAS Discovery, 2017; Winter et al, SLAS Discovery, 2019).
The high-throughput MALDI-TOF workflows developed to date typically monitor the substrates and products of in vitro biochemical reactions using recombinant proteins as enzymes (Ritorto et al, Nature Communications, 2014; De Cesare et al, Cell Chem Biol, 2017).
However, human primary and iPSC cell-based assays are increasingly being deployed in the early phases of drug discovery to better recapitulate human biology at scale, and ultimately improve the translation of screening hits into the clinic. A detailed characterisation of the cell models deployed in these assays is required. The aim of this research project will be to develop MALDI-TOF screening workflows for primary cell-based assays and methods for the validation of human relevant cell models that can be run at sufficiently high throughput to be applied in an industrial screening environment.
Initially, to develop expertise with MALDI-TOF technology and screening workflows, the student will develop a biochemical assay on the Bruker Rapiflex system and screen a validation set of small molecule inhibitors or activators. Once basic workflows are well established the second phase of the project will be to develop assays using primary human cells, with a focus on immune cells such as macrophages or T-cells which are of particular interest to GSK as they are targets for anti-inflammatory drugs and drugs in immuneoncology. Aspects of cell-based technology development such as workflows for adherent vs suspension cells, and the robustness and reproducible of the signal between cell batches or donors will be explored. Having established sample preparation and analysis methods for these primary immune cell populations, further work to define and develop peak ‘signatures’ that characterise fit for purpose cell batches will be conducted. Using these signatures, we hope to develop methods to compare cells from batch to batch or compare signatures for model cell lines to those of primary of iPSC derived cells. Overall the work will further our understanding of the cellular assay capabilities of high throughput MALDI-TOF, particularly in the immune cell space.
The student conducting this work will develop a solid understanding of MALDI-TOF techniques, and gain significant expertise in the application of MALDI-TOF for biochemical and cellular assays and screening workflows in formats relevant to drug discovery.
It is anticipated that the student will spend a significant portion of time after year 1 at GSK Stevenage, working with GSK scientists, gaining experience in an industrial setting and transferring methods onto the large scale automated MALDITOF platform (propose all of year 2).
The student conducting this research will build a solid understanding of assay design principles, requirements for small molecule screening and challenges when scaling and miniaturising human disease relevant biology. This will be a joint project with GSK, one of the CDT industrial partners who will provide 50% funding and work will be conducted both at Newcastle and GSK throughout the studentship.
Training & Skills
The student will be part of a multi-disciplinary and diverse research group at Newcastle University and will spend considerable time during the PhD at the Stevenage site of GSK. The student will be part of the CDT cohort and receive training in medicinal chemistry and drug discovery. She/he will also have access to the PhD programme which provides training in Research and Transferable skills.
The student will be trained in a wide range of techniques including state-of-the-art mass spectrometry, data analysis, enzyme drug discovery assays, primary cell culture and highthroughput drug discovery in an academic and a pharmaceutical industry setting. GSK will provide training in the latter.
Professor Matthias Trost
[Email Address Removed]
+44 (0)191 2087009
How to Apply
You must apply through the University’s online postgraduate application system
You will need to:
• Insert the programme code 8207F in the programme of study section
• Select ‘PhD in Molecular Sciences’ as the programme of study
• Please include the studentship reference code that you are applying for in the studentship/partnership reference field (codes are outlined in the individual projects found on the MoSMed website: research.ncl.ac.uk/mosmed/)
• Attach a CV and cover letter. The cover letter must state the title of the studentship, quote the relevant reference code and state how your interests and experience relate to the project
• Attach degree transcripts and certificates and, if English is not your first language, a copy of your English language qualifications
• Email a copy of your CV and cover letter to: [Email Address Removed] confirming the project that you have applied for