Location: Steinbeck 1

Zoltan Takats, PhD Imperial College Professor Takats has obtained his PhD from Eötvös Loránd University, Budapest, Hungary. He has worked as a post-doctoral research associate at Purdue University, Indiana, USA. After returning to Hungary, he served as Director of Cell Screen Research Centre and also as Head of Newborn Screening and Metabolic Diagnostic Laboratory at Semmelweis University, Budapest. Professor Takats was awarded the Starting Grant by the European Research Council in 2008 and he subsequently, became a Junior Research Group Leader at Justus Liebig University, Gießen, Germany. He moved to the United Kingdom in 2012 and currently works as a Professor of Analytical Chemistry at Imperial College London. Professor Takats has pursued pioneering research in mass spectrometry and he is one of the founders of the field of ‘Ambient Mass Spectrometry’. He is the primary inventor of six mass spectrometric ionization techniques and author of 78 peer reviewed publications. He was the recipient of the prestigious Mattauch-Herzog Award of the German Mass Spectrometry Society and the Hungarian Star Award for Outstanding Innovators. He is the founder of Prosolia Inc, Medimass Ltd and Massprom Ltd, all companies pursuing analytical and medical device development.

Objectives

The objective of the workshop is to identify and overcome potential roadblocks preventing mass spectrometry-guided surgery to become routinely used in the clinical interventional world. The workshop will focus on the side-by-side comparison of existing technology and clinical needs, as also on the embedding of the technology into existing healthcare settings and regulatory aspects. The workshop is envisioned to outline the strategy for the clinical translation of these technologies.

Summary

The need for in-situ, real-time tissue identification has been dramatically increasing with the development and deployment of robotic and other high-precision surgical approaches. While surgical mass spectrometry techniques have been continuously developed, published and demonstrated in human surgical environment, none of these approaches have reached regulatory approval and routine application in surgery. We are planning to use the meeting to understand where the roadblocks are and how we can get around them to deliver the technology for patients and healthcare professionals equally needing it. The workshop is envisioned to map out the current technology landscape and identify the strengths and weaknesses of each solution together with their respective low-hanging fruit applications as the first main topic. This mapping exercise will be followed by discussing the embedding of the approach both into existing oncology and clinical diagnostic systems. The approach is expected to change how interventional cancer care (and potentially a few other treatments) is delivered, hence it is of utmost importance to understand this aspect from the point of view of pathologists, oncologists and medical imaging professionals. Furthermore, the technology will also be the forerunner of a universal, mass spectrometry-based diagnostic system, bringing LC-MS and MS imaging experts to the table. Beyond the embedding of the technology, we are also keen to discuss the regulatory and health economic aspects of the approach in the third part of the workshop.

Syllabus/Topics

• Surgical mass spectrometry methods – strengths, weaknesses, applications and future perspectives
• Embedding of technology into healthcare systems. Interactions with oncology, pathology, imaging and clinical chemistry. Synergism with other MS-based diagnostic technologies.
• Regulatory and health economics aspects

Location: Steinbeck 2

Margret Thorsteinsdottir, PhD University of Iceland Professor in Pharmaceutical Analytical Chemistry at the Faculty of Pharmaceutical Sciences, University of Iceland and R&D Director of ArcticMass LTd, Reykjavik, Iceland. Dr. Thorsteinsdóttir received her PhD from Uppsala University, Sweden in 1998. From 2000 to 2009 she was the managing director of Bioanalytical Laboratories at deCODE Genetics, Reykjavik, Iceland. She has extensive experience in development of analytical methods for metabolite profiling and quantification of clinical biomarkers in various biofluids utilizing chemometrics with the goal of improved clinical management of patients towards personalized patient care. Her current research interest includes studies of lipid metabolism in cancer cells and profiling plasma derived biomarkers for early detection of BRCA-related breast cancer. She is responsible for implementation of clinical mass spectrometry for support of diagnostics and therapeutic drug monitoring in collaboration with ArcticMass and the Landspitali University Hospital, Reykjavik, Iceland with major focus on quantitative targeted proteomics for clinical diagnosis. She is a principal investigator of the Icelandic Research Rannis projects, profiling metabolites for breast cancer diagnosis and search for novel biomarkers for early breast cancer diagnosis by metabolomics. Dr. Thorsteinsdóttir is a principal investigator for the Marine Biotechnology ERA-net project CYNOBESITY and the Horizon 2020 project MossTech, with the main task to isolate, identify and structurally characterize bioactive compounds from cyanobacteria, Icelandic mosses and liverworts. She is one of the founders of Females in Mass Spectrometry (FeMS), she is a vice-leader of the working group clinical significance and applications of (epi)lipidomics in the pan-European network, EpiLipidNET and vice-chair of the Nordic Metabolomics Society.

Finnur Eiriksson, PhD ArticMass

Objectives

The objective of the workshop is to provide an introduction into design of experiments (DoE) for clinical application with special focus on optimization of MS-based clinical assays. The workshop is focused on practical implementation of DoE and will demonstrate how method development of sample preparation and UPLC-MS/MS method for quantification of clinical biomarkers can become much more efficient by utilizing DoE.

Summary

Design of experiments (DoE) is an efficient tool for development and optimization of UPLC-MS/MS platform for quantification of biomarkers in complex biological matrices. The UPLC-MS/MS platform is composed of several processes which involve many experimental factors that need to be simultaneously optimized to obtain a true maximum sensitivity with adequate resolution at minimum retention time. DoE offers a practical approach for performing experiments in accordance with a predefined plan, modelling by empirical functions, and graphical visualization. Basic concept of DoE will be presented with emphasis on practical implementation of DoE which includes the three main stages, screening, optimization, and robustness testing. To demonstrate the cost-effective benefit of DoE, which allows the effect of variables to be assessed with only a fraction of the experiments that would be required by changing one-separate-factor-at-time (COST) approach, two case studies will be presented. The first case is optimization of sample preparation in bottom-up targeted protein LC-MS workflow using DoE. The second case is an optimization of a UPLC-MS/MS assay for clinical diagnostic and therapeutic drug monitoring of patients with adenine phosphoribosyltransferase (APRT) deficiency, which is an inborn error of purine metabolism. A polynomial model which corresponds to the objective of the case study is specified and an experimental design that supports the selected model is generated. Significant factors were studied via central composite design and related to responses utilizing partial least square (PLS)-regression. Both cases showed that DoE is an excellent tool for optimization of sample preparation for biological samples and UPLC-MS/MS quantification method for clinical biomarkers. A significant reduction of sample preparation time was achieved with increased yields for selected peptides and a reliable UPLC-MS/MS assay for simultaneous quantification of urinary 2,8-dihydroxyadenine (DHA) and adenine was optimized efficiently with DoE.

Syllabus

• Design of Experiments (DoE) – Get it right from the beginning
• Basic concept and assessment of DoE
• Optimization of sample preparation and UPLC-MS/MS clinical assay by DoE

Location: Steinbeck 3

Christopher Chouinard, PhD Clemson University I received my PhD from University of Florida in 2016, where I developed ion mobility-mass spectrometry (IM-MS) methods for steroids and vitamin D metabolites. I then worked as a post-doctoral research at Pacific Northwest National Laboratory, building Structures for Loss Ion Manipulations (SLIM) ion mobility instrumentation for application in metabolomics and proteomics. In 2018, I began my independent career as an Assistant Professor at Florida Institute of Technology. I have since moved to Clemson University in August 2022. Work in my research group focuses on ion mobility-mass spectrometry (IM-MS)-based methods and technology, including structurally selective reactions for improved characterization of steroids and other controlled substances.

Robin Kemperman, PhD Children’s Hospital of Philadelphia Robin Kemperman received his Bachelor's in chemistry from the HAN University of Applied Sciences in The Netherlands. Thereafter, he fulfilled his MSc and PhD in analytical chemistry at the University of Florida under the direction of Dr. Richard Yost. Currently, he works at the Children's Hospital of Philadelphia as Sr. Mass Spectrometrist in the Metabolic and Advanced Diagnostics Lab. Dr. Kemperman's work has covered a variety of aspects in mass spectrometry, including targeted analysis of steroids and ketone bodies using LC-MS/MS, bile acid, opioid, and glycan isomer separations using ion mobility spectrometry, and metabolomics High-Resolution MS. Dr. Kemperman is experienced in clinical MS-based validations and has presented his work at a variety of national and international meetings. Focusing on the future, he is interested in working on novel innovations for biomedical and clinical applications.

Objective

Attendees will learn the basic principles of ion mobility, benefits and challenges to routine implementation in the clinical lab, method development, and current applications.

Summary

Ion mobility-mass spectrometry (IM-MS) has become a cornerstone of biomedical analysis, with applications ranging from isomeric small molecule differentiation to the study of protein structure. Despite its advantages, IM-MS has yet to see routine implementation in the clinical lab due to challenges in quantitation, limited universal standards, data processing software, and reproducibility across different IM techniques/vendor platforms. This workshop will introduce common IM techniques and their operating principles, expanding upon the benefits of incorporating IM into conventional LC-MS/MS workflows and discussing its challenges. A discussion of method design and development will focus on how users might go about integrating IM into their existing (or new) assays. Finally, an overview of current applications (including metabolomics, lipidomics, and proteomics examples) will be provided.

Objective 1: Understand the basic operating principles of IMS and the differences between the different techniques (e.g., drift tube, traveling wave, FAIMS/DMS, etc.)
Objective 2: Recognize the benefits and limitations to incorporating IMS into conventional LC-MS/MS workflows in the clinic
Objective 3: Become familiar with method design and development and current/future applications

Syllabus

• Basic Operating Conditions of IMS: Electric field application, experimental conditions (temperature, pressure, gas composition)
• Different IMS techniques: Drift tube/traveling wave, field asymmetric/differential mobility, emerging techniques (i.e., TIMS, SLIM, cIMS, etc.)
• Applications: Current examples from metabolomics, lipidomics, and proteomics

Location: Colton

Melissa Budelier, PhD TriCore Reference Laboratories Dr. Budelier the Medical Director of Clinical Chemistry and Toxicology at TriCore Reference Laboratories and Clinical Assistant Professor of Pathology at the University of New Mexico. Her research interests are broadly focused on developing clinically useful, mass spectrometry-based assays to improve diagnosis and treatment of human disease. Her expertise are in Toxicology/TDM, assay development and validation, and protein quantification.

Jacqueline Hubbard, PhD, DABCC Hubbard Lab Consulting Jacqueline Hubbard received her BS degree in Biochemistry from the University of Vermont. She then earned her MS and PhD in Biochemistry and Molecular Biology from the University of California, Riverside (UCR). Following a one year postdoc at UCR, Dr. Hubbard completed a Fellowship in Clinical Chemistry at the University of California, San Diego Health. She is board certified in Clinical Chemistry by the American Board of Clinical Chemistry. In 2019, she took a position as an Assistant Professor in the Department of Pathology and Laboratory Medicine at the Geisel School of Medicine at Dartmouth and as the Assistant Director of Clinical Chemistry at Dartmouth-Hitchcock Medical Center. There, she focused on developing and validating drugs of abuse assays and SARS-CoV-2 serology testing. In 2022, she became the Laboratory Director at Three Rivers Diagnostics, a reference laboratory in Pittsburgh, PA. Her research focus still includes mass spectrometry method development and toxicology test interpretation.

Objectives

• Recognize the role of laboratory developed tests (LDTs) in patient care
• Describe the current regulatory landscape of LDTs
• Define the VALID act and discuss its potential implications to the practice of laboratory medicine and patient care

Summary

Laboratory Developed Tests (LDTs) play a critical role in patient care. They are developed by expert laboratorians to bridge gaps between patient needs and available FDA-approved assays. LDTs are used to help diagnose and manage treatment for a variety of health conditions. These tests undergo extensive validation prior to implementation and, within the United States, are federally regulated by CLIA. The proposed Verifying Accurate Leading-edge IVCT Development Act of 2022 (“VALID Act”) includes additional regulation of LDTs by the FDA. If passed, the VALID Act would limit LDT access and the ability to provide timely laboratory testing to providers and patients, resulting in significant care gaps.

This interactive workshop will provide an overview on what LDTs are, how they are validated and the current regulatory framework, highlighting papers published in the recent JMSACL special issue – Laboratory Developed Tests: Regulation, Assay Development, and Impact on Patient Care. We’ll also provide an overview on the status and potential implications of the VALID act. In the second half of the workshop, the audience will have a chance to share their opinions, questions, and concerns in a flipped classroom Q & A.

Location: Stevenson 2

	Robert Gurke, PhD Fraunhofer Institute for Translational Medicine and Pharmacology ITMP Robert Gurke received his diploma in chemistry at the Humboldt-University zu Berlin, Germany in 2012 followed by his doctoral thesis at the Technische Universität Dresden in 2016. After a short period as study director in a GLP-compliant bioanalytical company in Berlin he started working as research associate at the Institute of Clinical Pharmacology as well as the Fraunhofer ITMP in Frankfurt under the guidance of Prof. Geisslinger. Robert Gurke is head of the LC-MS analytics group in both institutions since 2021. Mr Gurke is performing LC-MS/MS analysis since starting his doctoral thesis and gained broad experience in the field of developing and validating methods for the determination of exogenous and endogenous small molecules in different complex matrices.
	Anne K. Bendt, PhD Singapore Lipidomics Incubator (SLING), National University of Singapore Anne K Bendt studied Biology focusing on marine biotechnology (Greifswald University, Germany), followed by a PhD in Biochemistry (Cologne University, Germany) employing proteomics and transcriptomics. Driven by her fascination for infectious diseases, she joined the National University of Singapore (NUS) in 2004 to develop lipidomics tools for tuberculosis studies. She is now a Principal Investigator at the Life Sciences Institute, NUS, focussing on translation of mass spec technologies into clinical applications, and serving as the Deputy Director of the Singapore Lipidomics Incubator (SLING) taking care of operations and commercialization.
	Bo Burla, PhD Sling @ National University of Singapore I studied biology at the University of Zurich, Switzerland, and made my PhD in Molecular Plant Physiology in the lab of Prof. Enrico Martinoia, focusing on ABC transporters and comparative molecular phylogenetics. Subsequently, I became as researcher at the University Hospital of Zurich, where I was involved in establishing an LC/MS-based assay for the peptide hormone hepcidin and in projects studying Fabry Disease mechanisms. Biological processes, bioanalysis, clinical applications and the use of informatics to improve workflows have been my constant research interests. With this background I am now working as a senior researcher at the Singapore Lipidomics Incubator (SLING), heading our new data team that is focusing on developing workflows and software pipelines for the analytical data processing, QA/AC and exploration of the diverse lipidomics datasets generated by our lab.

Objectives

It is the objective of the workshop to provide an introduction into pre-analytical considerations for clinical application of MS-based analytics, with a special focus on lipids. However, these considerations are in many cases independent of specific analytes of interest and can hence be applied for polar metabolites as well as proteins. The workshop is focused on main hurdles to be considered when performing clinical research using LC-MS based determination of lipids namely: (I) pre-analytical sample handling, (II) frequent generation of patient samples as well as (III) overall cohort and study design.

Summary

Lipids are involved in a broad spectrum of functionalities in the organism and implicated in a variety of physiological and pathological processes. Changes in lipid levels are promising biomarkers for early diagnosis, prognosis of disease progression, or guidance for selecting promising therapeutic approaches. However, biomarker discovery in the field of lipidomics is a very challenging process since lipids require specific procedures regarding sampling (including selection of sample type and collection procedures, storage conditions and number of freeze-thaw cycles), sample preparation and analysis for reliable determination. As especially pre-analytical sample handling is a critical step for the reliable analysis of the lipidome, a close cooperation with the clinicians is necessary. This ensures following a highly standardized protocol for venous blood sampling to avoid several pre-analytical pitfalls potentially changing the lipid profile ex vivo. As collecting venous blood samples is very laborious for patients as well as clinicians, other ways for a more frequent sample collection are necessary. Therefore, capillary blood sampling using microsampling devices is an auspicious way to complement the strategy of analyzing venous blood-based samples taken in the clinics. Besides considering the right sampling strategy, the structured recording of sampling details and subject metadata, the overall planning of the study and also the cohorts to be included is of high relevance. All mentioned points have to be considered before starting a clinical research project applying lipidomics to generate high quality data making biomarker discovery possible.

Location: De Anza 1

Joshua Hayden, PhD, DABCC, FACB Norton Healthcare Joshua is currently the Chief of Chemistry at NortonHealthcare. He earned his PhD in chemistry from Carnegie Mellon University. He conducted postdoctoral research at Massachusetts Institute of Technology before completing a two-year clinical chemistry fellowship at University of Washington and 4 years as Assistant Professor at Weill Medical College. Joshua has special expertise developing and overseeing mass spectrometry assays in the clinical laboratory.

Objective

The objective of this workshop is to walk attendees through the process of acquiring/installing an instrument, developing their first assay, validating the assay as a laboratory developed test, and then ensuring adequate performance once live. This will be done using the presenters experience implementing a liquid chromatography tandem mass spectrometry (LC-MS/MS) assay to quantify buprenorphine in human urine. At the end of the workshop, attendees should be able to:

1. Justify the acquisition of an LC-MS/MS for clinical use
   
2. Explain the major steps involved in developing a small molecule LC-MS/MS assay
   
3. List the required elements for clinical validation of a small molecule LC-MS/MS assay
   
4. Propose the steps necessary to ensure accurate results once live on a validated LC-MS/MS method

Summary The continuing expansion of mass spectrometry into the clinical laboratory involves both expanding what testing can and should be done by mass spectrometry as well as expanding the number of labs implementing and utilizing existing/established assays. Unfortunately, too often the barrier of entry into mass spectrometry can be quite high. This workshop is designed to help walk attendees through the process of acquiring/installing an instrument, developing their first assay, validating the assay as a laboratory developed test, and then ensuring adequate performance once live (post-implementation monitoring). In particular, attendees will be given an overview of how these steps were done when the presenter setup and implemented a liquid chromatography tandem mass spectrometry (LC-MS/MS) assay to quantify buprenorphine in human urine in a College of American Pathologists (CAP) accredited laboratory. Special attention will be given resources that are available to assist labs undertaking the development and validation of clinical LC-MS/MS assays, resources such as published papers and CLSI guidelines. Practical factors such as sourcing consumables and necessary operating procedures will also be included. While the focus will be on implementing this specific assay, the goal is to share general best practices that were used that can be applicable to other assays.

Syllabus / Topics Covered

Presenting your business case and installing your instrument (0.5 hours, with intro)

Developing your assay (0.75 hours)
-Setting up chromatography
-Optimizing signal
-Selecting consumables (internal standards, QC, calibrators, mobile phase, etc)
-Finding your pitfalls

Validating your assay (0.75 hours)
-Reportable range
-Accuracy
-Precision
-Ion suppression
-Stability
-Testing your pitfalls

Post-implementation monitoring (0.5 hours)
-Daily quality assurance (SST, blanks, QC, etc)
-Data review
-Training/competency assessments