2021 SUMS-RAS Speakers
Florian Harking
PhD Student, NNF Center for Protein Research, University of Copenhagen
Automated and Ultra-sensitive workflows for Large-scale Quantitative Phosphoproteomics | View talk recording | view recording with Q&A
Phosphorylation is one of the most common post-translational modifications. It plays a key role for intra-cellular signal transduction and is known to be involved in numerous diseases. Current LC-MS/MS based workflows focusing on phosphorylation are aimed at processing samples with high sample amounts and low throughput. At the same time, a number of sample types, associated to the clinics, are more limited by their sample amount, but less by their number of individual samples. To address this discrepancy, significant efforts have been made in the last years regarding low-input amounts sample preparation and automation. Here, I want to display some of these efforts while presenting the benefits of using automated sample preparation for a more streamlined and reproducible workflow.
Bio: Florian Harking is currently a Marie Curie PhD student at the Novo Nordisk Foundation Center for Protein Research, University of Copenhagen. He is part of an European training Network on Palaeoproteomics. He has worked in the field of Proteomics for over four years, mostly in the setting of a contract-research company focused on proteomics applications for biotech and pharma. His work was focused on the application and development of targeted proteomics assays for (phospho-)proteomics and automation of sample preparation for high throughput assays. His current research interests include the adaptation of proteomics workflows for low input and partially degraded samples.
Sponsor: ReSyn Biosciences
Dr. Maureen A. Kane
Associate Professor; Executive Director, School of Pharmacy Mass Spectrometry Center, University of Maryland
Mass Spectrometry Imaging and Ion Mobility for the Study of Lipids in the Injured Brain | View talk recording
More than 1.7 million new cases of traumatic brain injury (TBI) occur annually in the United States. In addition to immediate mortality, TBI leads to high incidence of long-term disability and has been proposed as a major risk factor for the development of neurodegenerative disease. Our team recently demonstrated via lipidomics that there are substantial alterations in lysosomal membrane lipid composition in the cortex after traumatic brain injury. Alterations in cellular lipid composition and distribution after injury would be expected to have major effects on organellar biochemical and signaling activities, and thus on cell and organ function. Using the controlled cortical impact (CCI) model in mice, which provides a widely used and well-characterized method for investigating traumatic brain injury, we are interested in characterizing injury-related changes in lipid abundance as well as time-dependent changes in the spatial localization of lipids in respect to the injury site. Desorption Electrospray Ionization (DESI) has emerged as a powerful ionization technique for lipid imaging. DESI provides a softer ionization with less lipid fragmentation as compared to MALDI, however spectra can be complex due to endogenous adduct species that reflect the biological levels of Na+ and K+, which can change after injury. As DESI signal is highly dependent on salt, we modified our sample preparation by uniformly depositing sodium salt directly on top the tissue prior to imaging to shift adduct formation and enable the ionization of a greater diversity of lipid species. The addition of salt shows a higher signal for basic lipids by DESI but has also allowed the ionization of neutral lipids. It is of particular interest in the case of CCI, where we observe significant changes in these species after the injury. We also show the advantage of using ion mobility (IMS), in particular the use of a Cyclic IMS, which increases greatly the separation available for the identification of isobaric lipids.
Bio: Maureen A. Kane, Ph.D., is an Associate Professor in the Department of Pharmaceutical Sciences at the University of Maryland, School of Pharmacy and Executive Director of the University of Maryland, School of Pharmacy Mass Spectrometry Center. She received a B.S. in Chemistry from Canisius College (Buffalo, NY), a Ph.D. in Analytical Chemistry from the University at Buffalo, State University of New York, and completed postdoctoral training at the University of California, Berkeley. The Kane Lab focuses on elucidating disease mechanisms towards identifying new therapeutic targets and informing on mechanism(s) of action of current drug candidates using a combination of mass spectrometry-based techniques including quantitative LC-MS/MS, targeted metabolomics, lipidomics, proteomics, and mass spectrometry imaging. Efforts in the Kane lab also include the discovery and characterization of biomarkers for various applications. Dr. Kane’s work has resulted in over 150 peer-reviewed publications.
Sponsor: Waters
Dr. Gerald Larrouy-Maumus
Senior Lecturer, Imperial College
Deciphering Mechanisms of Immunometabolism in Eukaryotes and Drug Resistance in Bacteria using Extracellular Flux Analysis and 13C Stable-Isotope Tracing | View talk recording
Bioenergetics and metabolism are intimately connected. However, how those two can be executed and integrated can represent a challenge. Here, we will discuss the experimental design, analytical methods and interpretation of the date generated for two examples taken from our laboratory. The first one will uncover the changes in metabolism upon of LPS stimulation of macrophages. The second will focus on how Seahorse bioenergetics combined to flux analysis helped to decipher the mechanism by which Mycobacterium abscessus, a pathogenic mycobacteria, is so efficient in tolerating antimicrobials upon stressed by trace elements, such as copper, which is found in its environment.
Bio: Knowing that the mycobacterial cell envelope is one of the first links in the host-pathogen cross-talk, Dr Gerald Larrouy-Maumus did a Ph.D, in Toulouse (France), in order to study the biogenesis of the mycobacterial cell wall, especially the identification of the glycosyltransferases potentially involved in the biosynthesis of (lipo) polysaccharides which constitute the Achilles’ hill of the mycobacterial cell-envelope. As a lecturer in Molecular Microbiology at the MRC-CMBI, Imperial College, his laboratory explores deciphering the environmental adaptation of Mtb within the host. Mainly Metabolomics, and Transcriptomics, Proteomics, and Lipidomics are used as tools for the read-out of the first steps in this adaptation. Effectively, the success of Mtb as a pathogen partially results from its capacity to invade, survive and persist within intracellular phagosomes and extracellular sites in many host tissues. Throughout the cycle of infection, Mtb encounters and survives in a variety of harsh environments in the human body including nutrient-poor, acidic, oxidative, nitrosative and hypoxic niches. Very little is known about the molecular mechanism and kinetics of adaptation of Mtb during the first stages of infection within the host. Deciphering these mechanisms in such defined environments is crucial to understanding the physiology of Mtb within the host and can also inform on us why Mtb is such an efficient intracellular pathogen. The findings will potentially lead to the discovery of new drug targets and have a better understanding on resistant bacteria in context of the host.
Sponsor: Agilent
John Valliere-Douglass
Scientific Leader & Mass Spectrometry Core Group Leader, SeaGen
Merging Mass Spectrometry and Analytical Assay Data Sets to Streamline Biotherapeutic Protein Characterization and Enhance Product Understanding
Bio: John Valliere-Douglass is biotechnology professional with 25 years of industry experience in mass spectrometry and biotherapeutic protein characterization. He has authored over 30 publications on various aspects of mAb and antibody-drug conjugate (ADC) characterization and is a former co-chair for the CASSS mass spectrometry symposium. John is currently employed as a Scientific Leader at Seagen Inc. where he leads a core mass spectrometry group in the Process Development organization and serves as a cross-discipline key opinion leader that champions and establishes goals that position Seagen as a continued global leader in the development of oncology therapeutics.
Sponsor: Protein Metrics
Dr. Sebastian Virreira Winter
Chief Technology Officer, OmicEra Diagnostics
Biomarker Discovery in Body Fluids using High-throughput MS-based Proteomics | View talk recording
Bio: Dr. Sebastian Virreira Winter studied Molecular Biomedicine at the University of Bonn and holds a Ph.D. in immunology from the Humboldt University in Berlin. During his PhD, he focused on studying the innate immune response to infections in the lab of Prof. Arturo Zychlinsky at the Max Planck Institute for Infection Biology, Berlin. Afterwards, he joined the laboratory for Prof. Matthias Mann at the Max Planck Institute of Biochemistry for his postdoc. During this time, he developed a new isobaric labeling reagent – the EASI-tag – and co-developed MaxQuant.Live, a software enabling customized acquisition schemes and real-time monitoring of MS runs on Thermo Fisher Orbitrap mass spectrometers. In addition, he established new workflows for high-throughput urinary proteome profiling and discovered new biomarkers for Parkinson’s disease in CSF and urine in collaboration with the Michael J. Fox Foundation for Parkinson’s disease. Sebastian is a co-founder of OmicEra Diagnostics, where he currently serves as the Chief Technology Officer.
Before starting his PhD, Sebastian’s research activities took him to the University Clinic of Bonn, the Bayer HealthCare AG and the Whitehead Institute of Biomedical Research. He co-authored more than 10 peer-reviewed scientific publications and is an alumnus of the elite German Academic Scholarship Foundation and the Boehringer Ingelheim Fonds.
Sponsor: Evosep