Advanced Quantitative Mass Spectrometry Principles for Clinical Diagnostics Session 1 : Part 2 How to Achieve Lower Quantification Limits
Russell Grant, PhD
Labcorp
Dr. Grant earned a first-class honors degree in Industrial Chemistry from Cardiff University and a PhD in Chromatographic and Mass Spectrometric technologies from the University of Swansea, Wales, United Kingdom. He continued his scientific training in various industrial settings, which have included senior scientist at GSK, Principal scientist at Cohesive Technologies, Technical director at Eli Lilly, and Director of Mass Spectrometry at Esoterix Endocrinology. Dr Grant is currently the Vice President of Research and Development and co-discipline director for Mass spectrometry at Labcorp. Dr Grant has pioneered the use of direct injection technologies, chromatographic systems multiplexing, microsampling, utility of automation, and other new analytical platforms in direct patient care. His research goals are focused upon improvements in speed, sensitivity, and quality of liquid chromatography with tandem mass spectrometric (LC-MS/MS) analytical systems and assays. Dr Grant has been awarded 100 patents and received both the MSACL Distinguished contribution award and ASMS AL Yergey “Unsung Hero” Award in 2024 for his contributions to Clinical Diagnostics using Mass Spectrometry.
Note: This is Part 2 of a two part session.
The first session will be given as two 90-minute linked vignettes and will explore established and novel approaches to definitively improve analytical measurement performance - with the end goal of improving an assay LLOQ. The answer isn't necessarily buy a newer mass spectrometer, it is attention to details and maximizing each step of the process. This one-hour session will be presented in three modules. The first module, Foundations, will focus on assessment and improvements to pipetting techniques, control of adsorptive losses and use of surrogate IS studies to assess imprecision reduction during assay development. The second module, Formulation, will focus on system cleanliness, the fallacy of S:N, LC system dead-volume and flow rate implications, ionization efficiency enhancements, source and optics fouling, the value of signal summing + scheduled MRM and quadrupole resolution settings.
The third module, Finesse, will tie these concepts and practical solutions together to demonstrate their use for assays that have been developed, validated and deployed. Particular focus will be made to detailed establishment of “on-column” amount, controlling adsorptive losses throughout the assay, 2D-LC to decouple selectivity and sensitivity variables, selectivity and stoichiometry of reagents in worst case specimens, the consideration that while counter counter-intuitive, more degrees of selectivity (analyte losses) may be optimal for an assay
Take Home Pearls:
- How to practically improve an assay performance as defined by lower levels of quantification
- How to enhance the throughput of an LC-MS/MS assay by paying attention to the little details
- How to enhance signal responses in a mass spectrometer without sacrificing quality"
Grace van der Gugten, B.Sc. Chemistry Alberta Precision Laboratories
Advanced Quantitative MS Principles for Clinical Diagnostics Session 1 : Part 1 How to Achieve Lower Quantification Limits
Russell Grant, PhD
Labcorp
Dr. Grant earned a first-class honors degree in Industrial Chemistry from Cardiff University and a PhD in Chromatographic and Mass Spectrometric technologies from the University of Swansea, Wales, United Kingdom. He continued his scientific training in various industrial settings, which have included senior scientist at GSK, Principal scientist at Cohesive Technologies, Technical director at Eli Lilly, and Director of Mass Spectrometry at Esoterix Endocrinology. Dr Grant is currently the Vice President of Research and Development and co-discipline director for Mass spectrometry at Labcorp. Dr Grant has pioneered the use of direct injection technologies, chromatographic systems multiplexing, microsampling, utility of automation, and other new analytical platforms in direct patient care. His research goals are focused upon improvements in speed, sensitivity, and quality of liquid chromatography with tandem mass spectrometric (LC-MS/MS) analytical systems and assays. Dr Grant has been awarded 100 patents and received both the MSACL Distinguished contribution award and ASMS AL Yergey “Unsung Hero” Award in 2024 for his contributions to Clinical Diagnostics using Mass Spectrometry.
The first session will be given as two 90-minute linked vignettes and will explore established and novel approaches to definitively improve analytical measurement performance - with the end goal of improving an assay LLOQ. The answer isn't necessarily buy a newer mass spectrometer, it is attention to details and maximizing each step of the process. This one-hour session will be presented in three modules. The first module, Foundations, will focus on assessment and improvements to pipetting techniques, control of adsorptive losses and use of surrogate IS studies to assess imprecision reduction during assay development. The second module, Formulation, will focus on system cleanliness, the fallacy of S:N, LC system dead-volume and flow rate implications, ionization efficiency enhancements, source and optics fouling, the value of signal summing + scheduled MRM and quadrupole resolution settings.
The third module, Finesse, will tie these concepts and practical solutions together to demonstrate their use for assays that have been developed, validated and deployed. Particular focus will be made to detailed establishment of “on-column” amount, controlling adsorptive losses throughout the assay, 2D-LC to decouple selectivity and sensitivity variables, selectivity and stoichiometry of reagents in worst case specimens, the consideration that while counter counter-intuitive, more degrees of selectivity (analyte losses) may be optimal for an assay.
Take Home Pearls:
- How to practically improve an assay performance as defined by lower levels of quantification
- How to enhance the throughput of an LC-MS/MS assay by paying attention to the little details
- How to enhance signal responses in a mass spectrometer without sacrificing quality"
EDUCATION:
Graduate School: University of Virginia (Chemistry), Charlottesville, VA
Clinical Chemistry and Laboratory Medicine Fellowship: University of Virginia, Charlottesville, VA
CLINICAL:
Laboratory testing in Clinical Chemistry, Toxicology, Hemostasis, and Endocrinology.
RESEARCH:
Liquid chromatography mass spectrometry assay methods.
Dustin Bunch, PhD, DABCC
Nationwide Children's Hospital
Dustin R. Bunch, is an Asst. Director of Clinical Chemistry & Co-Director Laboratory Informatics at Nationwide Children's Hospital. His research focuses small molecule analysis by mass spectrometry in a clinical setting and clinical informatics.
Grace van der Gugten, B.Sc. Chemistry Alberta Precision Laboratories
MS Quantitation and/or Screening of Monoclonal Antibody Therapeutics: the Good, the Bad, the Ugly
Paula Ladwig, MS, MT (ASCP)
Mayo Clinic
Paula M. Ladwig, M.S., MT (ASCP), is a Principal Developer with the Clinical Mass Spectrometry Development Laboratory, Department of Laboratory Medicine and Pathology at Mayo Clinic in Rochester, MN. She has over 15 years of experience in the development and validation of new mass spectrometry tests. Her interests include the implementation of therapeutic drug monitoring of monoclonal antibody therapeutics by mass spectrometry.
Session Overview
The clinical laboratory will have many roles as monoclonal antibody therapeutics (t-mAbs) expand: identifying potential interferences in routine immunoassays; developing new assays to differentiate a t-mAb from an endogenous, disease-causing plasma cell clone and monitoring therapeutic drugs for better patient outcomes and assessing loss of response to therapy. This session will provide an overview of mass spec techniques available to the clinical laboratory for t-mAb detection and quantitation along with their advantages and disadvantages. Finally this session will offer a few examples of hurdles in the implementation into the clinical laboratory setting.
Target Audience
This session is intended for clinical laboratory directors and pathologists, clinical technologists, IVD manufacturers, pharmaceutical scientists, and anyone interested in the mass spectrometry applications for therapeutic monoclonal antibodies especially those involved in development of new methods for t-mAb monitoring.
Needs Assessment
There are over 60 different therapeutic monoclonal antibodies (t-mAbs) approved by the FDA; used to treat a variety of diseases. The market for monoclonal antibodies is rapidly growing, with over 500 new t-mAbs in several stages of development. Laboratorians are quite familiar with the detection, screening, and quantitative therapeutic drug monitoring for small molecules as the methodologies have been well established, therapeutic ranges for many drugs have been defined, and the metabolic pathways for many small molecules have been elucidated in detail. However, this is not the case for the t-mAbs. The detection, screening, and quantitative measurement of these monoclonal antibodies require different technologies.
Mass spectrometry is an important tool in the field of t-mAbs; mostly because it is relatively simple and so versatile once you understand how to apply the basic principles, challenges and limitations of each different approach and instrumentation. This session will provide examples of approaches for mass spectrometry assay development for chimeric, humanized and fully human t-mAbs quantitation; from peptide by quadrupole MS to intact or subunit detection by time-of-flight or orbitrap MS.
Following the completion of this session, the participant will be able to:
1. Describe mass spectrometry techniques available for the assessment of monoclonal antibody therapeutics along with their advantages and disadvantages.
Financial Considerations for Purchasing a Mass Spectrometer
Joe M. El-Khoury, PhD, DABCC, FAACC
Yale School of Medicine
Dr. Joe El-Khoury is Associate Professor of Laboratory Medicine at Yale School of Medicine, Director of the Clinical Chemistry Lab and Fellowship program at Yale-New Haven Health. He is board certified by the American Board of Clinical Chemistry (ABCC) and a fellow of the AACC Academy. He currently serves on the Board of Directors of AACC, as well as Chair of the Committee on Kidney Diseases for IFCC. His research interests are pre-analytical errors, biomarkers of kidney disease, and liquid chromatography-mass spectrometry in the clinical laboratory.
This informal Practical Training session will include a presentation on Financial Considerations for Purchasing a Mass Spectrometer, as well as offer small group discussion and networking opportunities.
1. Describe the clinical and monetary benefits of purchasing a mass spectrometry system.
2. Identify different models for financing a mass spectrometry system purchase.
3. Participate in effective negotiations with vendors.
Grace van der Gugten, B.Sc. Chemistry Alberta Precision Laboratories
Deborah French, PhD, DABCC (CC, TC) UCSF
Taking Aim at Analytical Interference in LC-MS Without Shooting Yourself in the Foot
Zlata Clark, PhD
B.S. in Analytical Chemistry, Masaryk University, Brno, Czech Republic.
Ph.D. in Bioanalytical Chemistry, Brigham Young University, Provo, Utah.
Nearly three decades of HPLC, CE, CE-MS, and LC-MSMS method development and validation experience in academic, pharmaceutical, and clinical laboratory environments.
Introduction
The popularity of LC-MS/MS-based methods for clinical testing continues to rise. However, despite their superior analytical specificity, these methods may still suffer from interference affecting method accuracy and precision, and hence negatively impacting patient care.
The aim of this Practical Training Track session is to introduce the participant to the following:
Segment #1
After each respective segment, attendees should be able to:
Segment #1
1.List sources of guidelines for interference testing
2.Define analytical interference and identify its various sources
Segment #2
1.Describe the different types of experiments used for interference testing in LC-MS/MS
2.Explain why waiting until validation to test for interference is not be desirable
Segment #3
1.Discuss the role of internal standard in mitigating interference
2.Name parameters used for interference monitoring
Grace van der Gugten, B.Sc. Chemistry Alberta Precision Laboratories
No Middleware? No Problem. Using R and Shiny for Routine Review of QC Data and Other Quality Metrics
Dennis Orton, PhD, FCACB
Alberta Precision Labs University of Calgary
I graduated with a PhD from Dalhousie University in Halifax, NS, Canada in 2014 where I worked on developing and applying quantitative proteomics workflows for biomarker discovery. I then completed a Clinical Biochemistry Fellowship in Calgary, AB, Canada in 2016 before moving on to work as a Clinical Biochemist in the Fraser Health region in British Columbia, Canada. During this time I gained significant experience in using R and RStudio, writing numerous shiny apps to perform QC management and to streamline LC-MS/MS data workflows. In 2019, I moved to my current position in Calgary where I head the Mass Spectrometry testing facility for Alberta Precision Laboratories. Primarily focussed on toxicology testing, I have overseen a transition towards more endocrine testing and eliminated low-throughput GC-MS workflows in favour of targeted, MRM based LC-MS analyses. My research is focused on promoting LC-MS technologies and development of tools and workflows to bring targeted proteomics methodologies to routine clinical application.
Introduction: Review of quality control (QC) data in the clinical lab generally utilizes vendor-specific and costly middleware systems which may not be user friendly or display all desired information. Additionally, many instruments may not come with a middleware option, the middleware may be cost-prohibitive, or it may not allow off-site data review, making routine QC review fairly labour intensive or time-consuming. These issues are especially problematic in regions with de-centralized clinical testing networks or with multiple analyzer vendors, and review of QC data is often limited to rudimentary Laboratory Information System (LIS) functionality, which is generally not user friendly or intuitive to use.
Objectives: Design a user interface to allow streamlined QC data review and allow rapid multi-site and/or multi-analyzer QC comparisons using R.
Methods: This script employs R (version 3.6.1) and RStudio (version 1.2.1335) with packages shinydashboard, ggplot2, and tidyverse to visualize QC data with filters for date range, assay type, QC product, analyzer, and QC lot number. The data is obtained from an automated download containing all QC data in the LIS over a 24 hour period with a sample identifier, test name, verified date, analyzer result, expected mean and standard deviation, QC product as well as the QC lot number. The data is saved to a shared network drive with access restricted to regional supervisory and technical staff.
The R script is set to automatically import the previous 30 days of QC data and displays the running mean and standard deviation for each test using the applied filters. Using shinydashboard format, more or less data can be viewed by importing more data files or by applying date range filters. Optional filters include test name, analyzer name/type, and QC material, which allow users to assess assay performance down to the instrument level. Multiple tabs are provided to display data in tabular or graphical format, with options for data to be summarize by day, week, or month.
Results: This dashboard provides a method for streamlining QC data review from various analyzer types and vendors, across sites and lot numbers, all of which can be viewed remotely. This provides technical staff the opportunity to quickly get through monthly QC review, as well as identify analyzers which may be seeing shifts in QC running means between analyzers or lot numbers. Ready access to this data allows staff to get through routine QC review quickly, while also promoting better region-wide lab quality and inter-site continuity. Adaptation of this dashboard could also allow review of other quality metrics such as patient running means or hemolysis rates, provided this data is captured in the LIS and access to the raw data is available.
Conclusion: This is a simple, customizable tool that is able to compile QC data for review without the need for investment in expensive or complicated middleware products.