The recent increase in large molecule therapeutics has been accompanied by an increase in the use of
peptide and LC-MS assays to analyze such molecules. Even though LBA’s are still the gold standard for large-molecule quantification, LC-MS has many benefits, such as a broader linear dynamic range allowing for multiplexing. However, there are still a range of challenges associated with the use of LC-MS for these compounds.
During this panel discussion we discussed what clinical stage peoples’ assays are used in, reflecting on what this indicates for the use of the technique in drug development. We also discussed sample preparation techniques used, covering proteolytic digestion and immunocapture techniques, with our panelists providing advice on best practices.
Additionally, our panelists addressed the drawbacks faced when using LC-MS to analyze peptides and large molecules. Aside from the aforementioned topics, much more was covered, ranging from internal standard selection to the appropriate acceptance criteria to apply. With a wealth of experience, our experts provided great insight and advice on key issues.
What will you learn?
- Method development for large molecule LC-MS assays
- Validation criteria to apply
- How to address drawbacks faced when using LC-MS for peptide and large molecule analysis
- Criteria for technology selection for fit-for-purpose large molecule bioanalytical strategy
Who may this interest?
- Bioanalysts working with the quantitation of peptides and large molecules
- Bioanalysts wishing to understand how to validate protein LC-MS assays for use in GxP studies
- Protein drug bioanalysts using ELISA who would like to learn more about LC-MS techniques for protein and peptide bioanalysis
- Protein LC-MS bioanalysts wishing to learn more about immunocapture techniques
- Individuals with an interest in LC and/or MS
Erin Chambers, currently Director of the Technology Advancement Group and a principal scientist at Waters Corporation, has been in the bioanalysis field for over 20 years, and at Waters for almost 15. She started her career at Pfizer supporting DMPK and has also worked as an MS service engineer and in instrument development for Applied Biosystems. In her current role, Erin develops bioanalytical applications for large and small molecule therapeutics and endogenous compounds. In addition, she has responsibility for assessing new sample preparation, LC and MS products as well as providing both in-house and customer training. Her most recent focus has been on the bioanalysis of proteins and peptides. Erin has authored numerous papers and a book chapter on this topic. Her current research interests include Alzheimer’s disease and diabetes.
Since 2005, Jeffrey Duggan has held the position of Director of Bioanalysis and Metabolite Profiling at Boehringer-Ingelheim (CT, USA). He is responsible for of all of that site’s small molecule bioanalytical studies as well as its ADME and metabolite profiling work. Currently, Jeff’s group is responsible for developing, validating and running protein LC-MS/MS bioanalysis at Boehringer-Ingelheim. Jeff received his MS and PhD in biological sciences from the University of Illinois (IL, USA). His current interest and most recent publications are concerned with the validation of LC-MS/MS bioanalytical methods for proteins in regulated studies. Jeff was a key organizer of a recent important industry white paper and has authored two additional articles on this subject.
Dr Fabio Garofolo has been working in the pharmaceutical and analytical fields since 1989. Capitalizing on an extensive occupational background within his specialized field, he has also been involved in helping scientific non-profit organizations (AAPS, ASMS, CFABS, CVG, GBC, GCC and NBC) with the mission to promote discussion on the pharmaceutical practices. Dr Garofolo has progressed throughout his career in both the pharma/biotech R&D and CRO industries and is presently the Head of Research at Angelini Pharma (Italy). He has over 180 publications & presentations at international conferences. Dr Garofolo has also developed over 400 innovative methods, including the design and invention of three innovative bioanalytical approaches and is the recipient of the following Lilly awards: Achievement (2001); Global (2002); Emmerson (2003). His current interests include analysis of large molecules by hybrid LBA/LC–MS, HRMS, DBS, and LBA emerging technologies.–
Roland is an Expert Scientist, heading the department of ‘Large Molecule Bioanalytical Sciences 2’ at Roche Pharma Research and Early Development (pRED), Roche Innovation Center Penzberg (Germany). He is a pharmacist and graduated in 2003 with a PhD in experimental and clinical toxicology (Hans H Maurer) from Saarland University (Germany), followed by a post-doctorate at the University of Geneva (Switzerland) in the life science mass spectrometry group of Gérard Hopfgartner. Roland worked as a forensic toxicologist at the institute of forensic medicine of the Ludwig Maximilians University in Munich (Germany), prior to joining Roche in 2007.
Jianing Zeng is an associate director of the Bioanalytical Sciences and Selective Integration Department at Bristol-Myers Squibb in Princeton (NJ, USA), leading a group of scientists in the development of LC-MS bioanalytical assays to support small and large molecule drug development. She is also heavily involved in biomarker and immunogenicity analysis with LC-MS technology. For the past few years, Jianing has been an active member of the AAPS Bioanalytical Focus Group on Protein Bioanalysis by Mass Spec, contributing significantly to the widely acclaimed White Paper ‘Recommendations for Validation of LC-MS/MS Bioanalytical Methods for Protein Biotherapeutics’.