Method Development Challenges, Regulatory Findings, and Innovations
in Bioanalysis
5th Workshop on Recent Issues in Bioanalysis (5th WRIB)
Workshop Lectures
Session 1: Introduction to 2011 Hot topics and Recent Issues in Bioanalysis - Dr. Fabio Garofolo, VP Bioanalytical Services, Algorithme Pharma., Canada
- Follow up on "unresolved" 2010 White Paper discussion topics - Bioanalysis (2010)2(12)
- Need-to-Know from 2010 Major Bioanalytical Conferences: NBC, Land O' Lakes, BSAT-APA, PSWC/AAPS/FIP, EBF
Session 2: Important Method Development Challenges in Bioanalysis
- Tissue Bioanalysis and Special Matrices
- Dr. Stacy Ho, Sr. Scientist, DMPK & Pharmaceutics, Genzyme, USA
- Tissue Homogenization
- Extraction techniques
- Issues with different tissue matrix. Types of tissues that exhibit larger variability than others and severe matrix enhancement/suppression and how to solve them
- Challenges in the development and validation of tissue methods
- Case Studies: Accurate Measurement of Drugs in Various Tissues
- Use of surrogate matrix (buffer or diluents) for calibration and QC sample for special matrix
- Method Development Strategies to Enhance Sensitivity in LC-MS/MS
- Mr. Richard Hucker, Manager, Clinical Assay Group, Pfizer, UK
- Clean up/Extraction strategies
- Chromatographic strategies
- Ionization strategies
- Dried Blood Spots (DBS): "Revisit with a Year of Progress"
- Dr. John Dunn, US Bioanalysis Head, GlaxoSmithKline, USA
- Dr. Steve Michael, VP and CSO of Global Bioanalytical Services, Covance, USA
- Have you implemented DBS after last year session? If not, we will share some further considerations associated with the implementation of DBS
- Validation techniques and process implementation to achieve accurate bioanalytical results
- Major Validation Considerations for DBS Analysis Methods
- The latest developments and Insights on DBS
- DBS: drawbacks!!
- High Throughput Method Development approaches
- Mr. Richard Hucker, Manager, Clinical Assay Group, Pfizer, UK
- Automation and when it helps
- High throughput & high batch failure: Strategies to develop high throughput method that are also rugged and robust
- Recent Method development Issues Related to Biologics and Ligand-Binding Assay (LBA)
- Dr. Binodh DeSilva (Session Chair), Executive Director Bioanalytical Sciences-Biologics, Bristol-Myers Squibb, USA
- Dr. Russell Weiner, Executive Director - Clinical Development Lab, Merck Research Laboratories, USA
- Dr. Jean Lee, Scientific Director, PKDM, Amgen, USA
- Dr. Dominique Gouty, Senior Director Immunochemistry, Intertek, USA
- Dr. Montse Carrasco, Development Scientist, Bioanalytical Sciences, Genentech, USA
- LBA Free/Total: recent data and industry standards
- LBA new technologies available: pros and cons (AlphaLISA , Gyrolab, etc.)
- Antibody Interference in LBA: what to consider
- Best Approach to determine LBA experimental Design
- Multiplexing: pros and cons of multiplexing; advantages in performing multiple measurements; Multiplexed assays development, validation and implementation
- Stability Issues and how to fix them during Method Development
- Dr. Wenkui Li, Sr. Fellow, Laboratory Head Translational Sciences - DMPK/DMBA, Novartis Institutes for Biomedical Research, USA
- Long Term Stability/Instability Issues in plasma and blood
- Impact of metabolites on stability
- Analyte instability issues in blood vs. DBS
- How to design a reliable whole blood stability evaluation
- Stabilities with multianalytes
- Matrix stability assessments: %Initial vs. %Nominal
Session 3: Innovations in Bioanalysis: From DD to DD (Drug Discovery to Drug Development)
- Accelerator Mass Spectrometry (AMS) and Microdosing
- Mr. Graeme Young, Manager PTS-DMPK, GlaxoSmithKline, UK
- Will AMS be a new key technique in bioanalysis?
- Specific emphasis will be put on the first white paper on validation of AMS for IV
- Microdosing and its advantages for studying the behavior of drugs in humans: PK data with almost no risk of side effects (Phase 0)
- Radioisotope carbon-14 labeled drugs and AMS extreme sensitivity for detecting only microdose levels of the drug
- AMS: Now a more accessible tool for bioanalytical quantitation, from large instrumentation available only in few specialized laboratories to the development of smaller affordable spectrometers
- Results of the European Union Microdose AMS Partnership Programme (EUMAPP) & scientific, ethical, and financial advantages
- Recent advancements in Protein drug Quantitative bioanalysis by LC-MS
- Dr. Mohammed Jemal, Senior Research Fellow, Bristol-Myers Squibb, USA
- LBA vs. LC-MS or LBA & LC-MS:
- Use of immobilized ligand and immunoaffinity columns as preparative/enriching technique for LC-MS/MS quantitative analysis of protein and biomarkers
- LC-MS as an important orthogonal method to LBA to provide structural elucidation of antibody drug conjugates and fusion protein
- Use of mass spectrometry to confirm immunoassay results
- Innovative development in LC-MS technique that can approach the performance of LBA
- LC-MS/MS and absolute quantification of protein therapeutics and biomarkers
- LC-MS ability to determine therapeutic proteins structural changes that change their biological activity
- Challenge in validation of LC-MS method for protein quantification: lack of well characterized RSM, analyte-free matrix, use of surrogates matrices, lack of specific guidance
- Mass Spectrometry analysis of diagnostic peptide fragment vs. intact protein
- Quantification of antibody therapeutics by LC-MS/MS
- High resolution (HRes) accurate mass spectrometry for quantitative bioanalysis
- Dr. Mohammed Jemal, Senior Research Fellow, Bristol-Myers Squibb, USA
- Ongoing technological advances in HRes (QTOF) and appealing possibilities for quantification of drugs in bioanalysis
- The most recent applications of exact mass MS in bioanalysis discovery/development quantitation
- Triple quadrupole MRM vs. QTOF Accurate mass Full spectrum MS and MS/MS data: selectivity, sensitivity and linearity, signal-to-noise, acquisition speed & mass accuracy & spectral quality
- High Resolution MS: narrower mass windows & higher selectivity
- Important case studies
Session 4: Regulatory Findings from Audits/Inspections
- Issues with Calibration curve: Small and Large Molecules
- Dr. Surendra Bansal, Research Director Bioanalytical R&D, Non-Clinical Safety, Hoffmann-La Roche, USA
- Dr. Dominique Gouty, Senior Director Immunochemistry, Intertek, USA
- Important considerations on linearity, slope, intercept
- Slope variability from instrument to instrument: Calibration curve slope importance in LC-MS/MS and system cross-validation for same instrument brand and/or different instrument brand
- Evaluation of Parallelism and curve fitting in LBA
- LBA & Validation of large molecules: specificity, selectivity and non linear calibration
- Biomarkers: Findings for small and large molecules
- Dr. Stephen Lowes, Senior Vice President Scientific, Advion BioServices, USA
- Evaluation of "fit-for-purpose" validations
- Method Qualification vs. Validation for Nonclinical Biomarker
- Industry trends
- Case studies and practical approaches
- Focus on Internal Standard (IS): "The year of the Internal Standard"
- Dr. Keith Gallicano, Director, Biopharmaceutics Operations, Watson Laboratories, USA
- Guidance in IS criteria
- Recent 483s on IS variation
- IS effects on accuracy and precision
- Acceptance criteria for the variation of IS for standard/QC vs. samples
- Trend analysis by using IS during investigations
- Incurred Samples Reanalysis (ISR) and IS: ISR Investigations and Closeout
- Do you need a stable labeled IS for each analyte/metabolite?
- Recent Experience with Regulatory Agency inspections: industry perspective
- Mr. Corey Nehls, Executive Director Bioanalytical Labs, PPD, USA
- Working together to improve quality
- Issued 483s: Opportunities to Discuss Science and Address Misunderstandings
- Importance of applying sound scientific principles throughout the bioanalytical activities
- FDA "Current thinking" through the 483 process: novel findings that introduce new expectations
- Issues in assay cross-validation/method transfers
- Mr. Peter van Amsterdam, Head of Global Bioanalytics, Abbott, The Netherlands
- Cross-validation guidance for same method used in several countries
- Partial validation vs. full validation
- Harmonization for Assay Transfer
- Selectivity evaluation for methods used to analyze global multi-center studies: use blank matrices from different countries
- OTC interference evaluation for methods used to analyze global multi-center studies
- New Findings on unstable metabolites: the generic drug issue
- Dr. Olivier Le Blaye, Inspector - Clinical Trials Inspection Unit - French Health Products Safety Agency - AFSSAPS - France
- Impact of metabolites on data accuracy in bioequivalence (BE) studies
- Examples of various problems found during inspections
- Clopidogrel, perindopril glucuronides, and lactones
- Long Term (LT) stability with Co-administered Drugs
- Dr. Eric Woolf, Senior Director, Merck Research Laboratories, USA
- Impact of Co-Administered Medications on Bioanalysis
- Three types of studies in which "co-meds" are an issue: DDI, combo product, and diseased population
- Approaches to handle studies with co-administered medications
- Dealing with oncology studies and their large mixture of co-administered drugs
- Stabilities for drugs that are marketed as a co-administered
- Fixing Hemolysis issues during Method Development
- Dr. Daniel Tang, Vice President of Bioanalysis and DMPK, Frontage Laboratories, China
- Potential impact of hemolysis on bioanalysis
- Accuracy and sensitivity
- Repeatability - ISR
- Stability
- Strategy to handle the hemolyzed samples in bioanalysis
- Assessing impact of hemolysis in pre-study method development/validation: what, how and issues
- Assessing data validity of each hemolyzed sample during sample analysis run: proposed strategy
- Data report for hemolyzed samples
- Regulatory/Quality Findings and their Impact in a Regulated Bioanalytical Environment
- Dr. Mario Rocci, Executive Vice President, ICON, USA
- While compliance with appropriate regulations is non-negotiable, the form that such compliance takes should make operational sense to the laboratory
- Quality findings should always be evaluated in terms of the added value they bring to the organization
Session 5: Upates and Findings from Regulatory Agencies and Technical and Scientific updates on Harmonization of Bioanalysis Guidance
- Presentations from Regulatory Agencies & Health Authorities worldwide
- Dr. CT Viswanathan, Associate Director, Division of Scientific Investigations, US FDA
- Dr. Brian Booth , Deputy Director, Office of Clinical Pharmacology, US FDA
- Dr. Olivier Le Blaye, Europe-EMA representative /Inspector France AFSSAPS
- Mr. Andrew Adams, Director, Health Canada Therapeutic Product Directorate
- Dr. Noriko Katori, Senior Researcher, Japan MHLW-NIHS
- Ms. Ariadna Cristina Gomes Barra, Head Bioequivalence Coordination, Brazil ANVISA
- Updates: Regulations, guidance, revisions
- Recent Findings
- Progress in the FDA and EMA collaboration in harmonizing their bioanalytical guidance and recommended practices to prevent conflicting approaches prior to their release
- Exhaustive update on global harmonization of the bioanalytical method validation and sample analysis guidance
- Parallel Activities of the Global Bioanalytical Consortium (GBC)
- Dr. Fabio Garofolo, VP Bioanalytical Services, Algorithme Pharma
- Continuing, through international harmonization teams, its scientific mission to merge existing or emerging bioanalytical guidances to create one, unified consensus document that can be presented to the regulatory bodies/health authorities in various countries
Session 6: PANEL DISCUSSION and 2011 WHITE PAPER preparation
Questions to be addressed to all the REGULATORY AGENCIES and INDUSTRY EXPERTS:
- Alternate detectors (AMS, High Resolution MS, ICPMS)
- Which guidance to follow for method validation and sample analysis?
- It seems that different auditors interpret the guideline in different ways:
- Is it possible to create consistency amongst inspectors?
- Can 483s be posted on a website for discussion?
- Batch failure:
- What is an acceptable level of batch failure 10%, 20%,...50%...more?
- Tissue analysis:
- What are the tips & tricks to succeed in tissue analysis?
- Whole blood stability evaluation:
- What implication does it have when the concentration result of whole blood QCs is significantly different than nominal concentration (e.g.: drug bound to protein or red blood cells)?
- What are the industry standards for this evaluation?
- Effect of counter-ion anticoagulants:
- Different agencies' approach?
- Is it real or just a matrix effect when we analyze multiple plasma lots?
- Differences in slopes of the calibration curves on different LC-MS/MSs:
- Is there any impact on the data?
- Chromatograms integration:
- When is manual integration accepted?
- Systems cross-validation:
- Is it needed and if yes in which cases?
- Variability of the internal standard (IS) in analytical and abnormal IS:
- Do we need to establish acceptance criteria for IS?
- Is IS trend analysis a new way to evaluate method reliability?
- Re-injection vs. re-analysis vs. non-reportable values:
- What is the industry standard?
- Stability issues in bioanalytical methods validation and the definition of "fresh":
- Is it necessary to use fresh QCs for stability assessments (not just calibrators)?
- Matrix stability for co-formulated drugs and co-administered drugs:
- Where do we stop testing?
- What are the Agencies' recommendations?
- Hemolysis
- What if the method is not insensitive to hemolysis?
- Can we still assign samples as "Not Reportable" or do we have to redevelop an "hemolysis-insensitive" method?
- "fit-for-purpose" validations
- Clarification and definition?
- Method transfers and cross validations
- What is the industry standard?
- Method Development data
- Can these data be integral part of an inspection/audit?
