A Researcher Live Series

Applications of Mass Spectrometry

 

20-28th July 2022

#MassSpectrometryLive

About this series

Mass spectrometry is a tool for measuring the ratio of mass to charge in an ion, with the results shown in an intensity plot known as a ‘mass spectrum’. Mass spectrometry aids in the analysis of proteins, metabolites, weight and chemical characteristics of molecules. A number of fields use mass spectrometry to analyse results and derive scientific conclusions. This includes carbon and radioactive dating, proteomics, metabolomics, clinical diagnostics and more.

In this series, we invite experts to discuss their work in applying mass spectrometry.

Programme

Hear from experts in the field

Join our speakers for discussions and Q&A about their latest research and discoveries

 


Mariya_Misheva

A new method for the analysis of short-chain fatty acids (SCFA) and other polar metabolites in microbiome-related samples by ion-exchange chromatography-mass spectrometry (IC-MS) with Dr Mariya Misheva, University of Oxford

The role of microbially-derived polar and ionic metabolites, e.g., short-chain fatty acids (SCFA), is of considerable interest in studying microbiome-host relationships in the context of health and disease. Detection and quantification of SCFAs in biological matrices, such as blood, urine and faecal samples, remains challenging in part due to low selectivity, high compound polarity, low abundance and volatility.

IC-MS enables separation, detection and quantification of biologically relevant SCFA and their isomers, including a wide range of additional organic acids, relevant to biological sample including blood plasma and faecal extracts. Therefore, IC-MS methods can be used for analysis of microbiome-related samples in both targeted and untargeted metabolomics experiments. Importantly, IC-MS offers minimal sample preparation without the need for derivatisation is needed. Thus, IC-MS can successfully detect and quantify microbiome-related volatile and non-volatile metabolites.

About the speaker

Dr Mariya Misheva is a PDRA in Microbiome-related Metabolomics in the group of Professor James McCullagh, Director of Mass Spectrometry Facility, University of Oxford. Her position is funded by the Oxford Centre for Microbiome Studies (OCMS) team, and her research focuses on developing new metabolomics methods and analysis pipelines to accelerate researching the role of the microbiome and its metabolome in health and disease. 

Mariya’s interest in mass spectrometry started during her PhD project where she used proteomics to investigate c-Jun N-terminal kinase (JNK1) post-translational modifications and protein complexes under basal and stress conditions. In the group of Valerie O’Donnell my work has focused on elucidating the role of lipids in innate immunity, more specifically eicosanoid production by microphages in inflammation. Lastly, my current position involves microbiome-related metabolomics projects with an emphasis on the gut microbiome metabolome and its role in healthy lifestyle and disease progression. 

Her career objectives are to increase her expertise in mass spectrometry and to use her experience to develop new tools, methods and protocols for solving complex biological problems. 


Aaron Bailey

Building a Versatile Intact Mass Toolkit to Improve Integrative Protein Structure Characterization for Drug Development and Basic Research with Dr Aaron Bailey, University of Texas 

Protein primary structure characterization is a basic requirement for understanding structure/function relationships in all application areas of protein engineering and basic research. Purified protein samples are mixtures of specific protein isoforms, or “proteoforms”, which are based on a common amino acid sequence with various possible post-translation modifications (PTMs) and/or sequence changes. Mass spectrometry, often combined with separation by liquid chromatography (LC−MS) is the predominant means for characterizing proteins, minimally achieved using a combination of two assays: LC−MS/MS peptide mapping analysis and LC−MS intact mass analysis. (...) To address issues of protein overcharging of unstructured proteins under acidic, denaturing conditions and sample heterogeneity (macro- and micro-scales) (...), we demonstrate the use of broadband isolation of entire charge state distributions of intact proteins followed by ion-ion proton transfer charge reduction. We term this "full scan PTCR" (fsPTCR). Using rapid denaturing size exclusion chromatography coupled to fsPTCR-Orbitrap MS and time-resolved deconvolution data analysis, we show a strategy for method optimization, leading to significant analytical advantages over conventional MS1. The increased sensitivity and dynamic range afforded by fsPTCR denaturing intact mass analysis has significantly improved our ability to confidently integrate the multi-assay LC-MS characterization efforts of a wide variety of protein samples.

About the speaker

Dr. Aaron O. Bailey is currently a Senior Research Scientist and Associate Director of the Mass Spectrometry Core Facility at the University of Texas Medical Branch. After graduate school, Dr. Bailey joined the Life Science Mass Spectrometry group at Thermo Fisher in San Jose, CA and worked closely with both Research & Engineering and Marketing groups by testing prototype HPLC and MS instrumentation and developing new methods for LC-MS characterization of intact proteins, including novel methods for in-line coupling of novel non-denaturing separations to non-denaturing mass spectrometry using newly compatible Orbitrap mass spectrometers. Dr. Bailey returned in late 2019 to academic research, joining the Mass Spectrometry Facility at the University of Texas Medical Branch in Galveston, Texas, USA. Under the leadership of Mass Spectrometry Facility Director, Dr. Bill Russell, Dr. Bailey continues to focus on developing new LC-MS technologies for protein characterization, leveraging combined efforts of bottom-up proteomics approaches and LC-MS/MS peptide mapping methods, as well as multiple LC-based protein separation strategies enabling intact mass profiling in both denaturing and native conditions.


Roopesh Krishnankutty (2)

Mass Spectrometry-based bottom-up proteomics and its applications in cancer research: an overview, with Dr Roopesh Krishnankutty, University of Edinburgh

The talk will present an overview on basics of mass spectrometry-based proteomics, different types of data acquisitions, basics of MS data analysis, software used for downstream data processing, data enrichment analysis as well as visualizations for data interpretation. In addition, it will give you an overview of how proteomics could be applied for cancer research by presenting the data obtained from a previously published research project. 

About the speaker

Dr Roopesh Krishnakutty is part of the Mass Spectrometry Facility at Institute of Genetics and Cancer (IGC), The University of Edinburgh, Scotland as a Senior Research Officer where he leads the proteomics-based research projects of the core facility. Dr. Krishnankutty has published many articles in ISI journals and has authored and co-authored many book chapters. His research interests are mainly focused on protein characterization using Mass Spectrometry-based Proteomics approach. Dr. Krishnankutty is actively involved in developing novel proteomics-based workflows to apply this technology in cancer research.


Gillian Taylor

Determing mass, why it is important and why it matters, with Dr Gillian Taylor, Teesside University niverrsUnive

Mass Spectrometry is a fundamental technique used across many science disciplines. Over recent years, instrumentation has helped understand organic residues in pottery, provenence of artefacts, animal faeces and much more. We will explore how mass spectrometry can help is undertand our past and how this can help link to our future

About the speaker

Gillian Taylor obtained a doctural degree on Bioarchaeological applications in Mass spectrometry. Her research interests mass spectrometry application and method development, currently focused on Magna – a Roman fort site in Northumberland. Gillian has published in a range of journals, including Antiquity, scientific reports and Journal of archaeological science.

Gillian taylor can be contacted at g.taylor@tees.ac.uk


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