Training Session T1 - Training Topic List
June 17, 2020, 11am to 4:30pm ET: Part 1
June 18, 2020, 11am to 4:30pm ET: Part 2

"Large Molecules Method Development by LBA: Solving Challenges with Innovative Approaches on Critical Reagents, Learning from Vaccine Assays, Multi-Domain Biotherapeutics, Emerging Technologies"

What’s new in Critical Reagents and impact on method development? What can we learn from Vaccine Assays experience? How Multi-Domain Biotherapeutics (MDB) are changing traditional method development strategies? How can Emerging Technologies (ET) help in method development?

View T1 Agenda

  • Dr. Christine Grimaldi,Head Biotherapeutic Bioanalysis Group,Boehringer Ingelheim
  • Dr. Rocio Murphy,Senior Principal Scientist Bioanalytical, Merck

Large Molecules by LBA Method Development Challenges & Emerging Technologies (ET)

What can we Learn from Vaccine Assays Experience?

What’s new in Critical Reagents and Impact on Method Development?


Topic DETAILS of T1 – LM Method Development by LBA

  • Dr. Christine Grimaldi,Head Biotherapeutic Bioanalysis Group,Boehringer Ingelheim
  • Dr. Rocio Murphy, Senior Principal Scientist Bioanalytical, Merck
Large Molecules by LBA Method Development Challenges & Emerging Technologies (ET)
  • Lesson 1
    Single Well Analysis ("Singlicate") in LBA: Learning from advanced & well-established pharma experience
    - Dr. Andrew Mayer, Group Leader Pharmacokinetic Assay Development, GlaxoSmithKline
    • Building on the 2019 White Paper in Bioanalysis Recommendations on ICH M10 for Single Well Analysis in LBA with novel practical case studies
      • 2019 Link:
      • 2012 - Consensus was reached that it is acceptable to run singlicate analysis if method robustness is proven
      • 2014 - It was clarified that Singlicate analysis should be based on the performance of the assay used
      • 2016 - Once the precision of the method has been demonstrated through validation, it is reasonable to move forward with a Singlicate approach for PK sample analysis
    • Single Well Analysis is now well-established in the industry.
      • Advanced training to ensure the correct application of single well analysis to Regulated Studies in LBA
    • Case studies
      • Acceptance Criteria used for submitted studies
      • Challenges in the approach and how to overcome them
      • Samples in singlicate and standards/QCs in singlicate
      • Masking of outliers and rules around such masking

  • Lesson 2
    Free & Total PK Assays Method Development and Validation: New Case study Learning from advanced & well-established pharma experience
    - Dr. Matthew Andisik, Assoc. Director Clinical Bioanalysis Group, Regeneron
    • Free PK Assays are always extremely challenging
      • Evolving science for Free & Total PK Assays
      • Free biotherapeutic
      • Free Target ligand
      • Bound: Mono- and/or bivalent complexes biotherapeutic/ligand
      • Total
    • Complexity of the dynamic binding equilibrium in vivo
      • Controlling multiple sources of perturbation
      • Strategies for reliable ex vivo quantification
      • Importance of developing the right LBA critical reagents
      • Best assay format design to measure free/total
    • Case studies on biotherapeutics practical method development for total/free analyte assessment

  • Lesson 3
    How are Multi-Domain Biotherapeutics (MDB) changing traditional LBA method development strategies? – Building on WRIB recommendations, new case studies and evolving trends
    - Dr. Seema Kumar, Assoc. Director DMPK, EMD Serono
    • Building on the 2014-2016 White Paper in Bioanalysis Recommendations on MDB PK Assays Strategies with novel practical case studies
    • 2014 Link:
      • "For the bioanalysis of MDB, multiple PK assays using several critical reagents may be required at an early stage in order to build an understanding of the moieties that impart clinical impact"
      • "Once an understanding has been established, measurement of only those relevant species can be performed moving forward."
      • "The stage of development dictates which analytes are important to monitor and the primary goal is to understand the moieties that are active and relevant to safety and efficacy"
    • 2016 Link:
      • "…it was concluded that when developing an assay for MDB, a complexity-based approach according to the therapeutic MoA should be used."
      • "It should be demonstrated that target binding is intact for the individual domains."
      • "Emerging multiplex platforms were encouraged even though they are not specifically designed for measuring MDB as part of a single assay with a limited sample volume if deemed appropriate."
    • Focused training on LBA method development for MDB
      • MDB currently in development
      • Complex mechanisms of action (MoA)
      • Evaluation during method development of domains each with a specific role or function
      • Multi domains epitopes with competitive binding
      • Development of specific Critical Reagents for MDB
      • Best assay format for MDB
    • Case studies: PK assay strategies MDB
      • What's best to measure?

  • Lesson 4
    Importance of QC Trending in Method Development, Validation and Sample Analysis for PK Assays by LBA: Building on WRIB recommendations, new Case Studies, lesson learnt and evolving trends
    - Dr. Mitra Azadeh, Assoc. Director Bioanalytics, Takeda
    • Building on the 2014 & 2015 White Paper in Bioanalysis Recommendations on QC in LBA with novel practical case studies
    • LBA performance & QC performance in
      • Method Validation
      • Pre-study method validation
      • In-study sample analysis.
    • Practical and focused training QC in LBA
      • Importance for Trending Analysis
      • Additional QC levels during LBA
      • Lot to Lot Consistency
      • Assay Drift Reasons & Troubleshooting
    • QC life cycle management
      • Methodologies and statistical tools to trend QCs
    • Regulatory concerns with QC qualification, Additional QC levels, QC trending, and maintenance of lot to lot consistency
    • Case studies on how to maximize the use of QC for developing robust LBA assay with consistent performance over time

  • Lesson 5
    Common Issue with LIMS based software for LBA support: Sharing pharma experience
    - r. Marcela Araya, Group Leader BioMedicine Design, Pfizer
    • Current industry concerns with Watson LIMS for LBA
      • Developed at first for PK support using Mass Spectrometry
      • Not completely adapted to LBA assay
      • Major problems during the method development stage
    • Common Watson LIMS questions from the LBA bioanalytical community
      • More active discussion needed within the community to share on LIMS issues and take actions
        • Watson LIMS response to industry concerns is somehow to slow
        • How to speed up Watson LIMS upgrades to better meet current industry standards in LBA
    • What kind of LIMS are Pharma/CRO using for LBA support beyond Watson LIMS?
      • In Regulated (Validation & Sample analysis)?
      • Method development?
      • ADA Assays?
    • Industry experience in using Softmax software vs Watson LIMS in LBA Method Development
      • Advantages with multiple assay
      • Extra flexibility for data analysis
      • Easy transitioning from Softmax in method development to Watson LIMS in Validation and Sample Analysis
    • Case studies: Difficulties to use LIMS tool efficiently for LBA support and possible solutions

  • Lesson 6
    Can Ella Platform be used for PK Assays in Regulated Bioanalysis? – Pros/Cons and Method development / validation challenges
    - Dr. Jasna Canadi Semadeni, Head Regulated Large Molecules Bioanalytics, Roche
    • Ella is a microfluidic cartridge-based LBA
      • Ella main applications have been mainly in the Biomarker field so far
      • What are the potentials of Ella for PK Assays?
      • Can Ella unique platform help facilitate drug quantification?
    • Importance of Emerging Technology that are starting to enter the bioanalytical lab
      • How is the industry handling these new technologies from a regulatory perspective?
      • How is Ella addressing different study needs for PK assays in a regulatory environment?
        • Efficiency
        • Sample handling/consumption
        • Automation
    • Challenges with Method Transfer from traditional to Emerging LBA platforms
      • Is Cross Validation needed?
      • See 2014 White Paper in Bioanalysis recommendations on LBA platforms cross-validation
      • "…cross-validation with an existing technology is necessary only if the change of platform occurs within a program or a study…"
      • 2014 Link:
    • Case studies on Ella application as Emerging Technology for PK Assays


What can we Learn from Vaccine Assays Experience?
  • Lesson 7
    Strategies in Vaccine Clinical LBA Development: What can the Bioanalytical Community learn from Vaccines LBA method development and validation?
    - Dr. Rocio Murphy, Senior Principal Scientist Bioanalytical, Merck
    • Development of LBA to define clinical endpoint in the assessment of new vaccines
      • Immunogenicity as surrogate of vaccine efficacy
    • Main characteristics of Vaccine Clinical LBA
      • Highly characterized
      • Well-controlled
      • Statistically supported
    • LBA Development & Optimization
      • Assay Robustness and Ruggedness
      • Stringent qualification and validation requirements
      • Ensuring long-term assay performance
      • Traditional ELISA and multiplex (Luminex, MSD)
      • Determination of assay LOD and LLOQ
      • Establishing QC ranges
      • Establishing proficiency panels for long term monitoring
    • Major challenges for the development of Vaccine Clinical LBA
      • Lack of appropriate reference standards
      • Lack of appropriate positive control samples
      • Need to mimic the antibody profile
    • Lessons learned from regulatory feedback on assay validation
    • Case studies on vaccine assay development including LBA selection and the use of Design of experiments (DOE) to determine optimal conditions for the assay

  • Lesson 8
    Scientific & Regulatory Differences/Similarities between Vaccine and Biotherapeutics LBA: Bioanalytical case studies on clinical testing of wanted and unwanted immunogenicity
    - Dr. Ivo Sonderegger, Assoc. Director Clinical Serology Strategy, Takeda
    • Similarities and differences between Vaccine & Biotherapeutics LBA
      • Same assay technologies
      • Major differences in
        • Goal of the clinical testing
        • Strategies applied
    • Biotherapeutics - Unwanted immunogenicity
      • LBA focuses on the detection of very small amounts of ADA
      • Impact on the safety and efficacy
      • Current practice: tiered approach
        • Screening assay
        • Confirmatory assay
        • Characterization: Titer and neutralizing assay
      • Established Regulatory Guidelines
    • Vaccine
      • LBA assess the desired immunogenicity of a vaccine.
      • No need for high assay-sensitivity
      • No need for the tiered approached
      • LBA need to describe
        • Desired immunogenicity of the vaccine
        • Optimally correlate with the efficacy of the vaccine
      • Stable long-term use of the assays
      • Limited published Regulatory Guidelines
    • Case studies for immunogenicity evaluation of a vaccine candidate will be used to clarify the difference between the two worlds

  • Lesson 9
    LBA for Vaccine, Development, Validation & Troubleshooting: Challenges with concomitant vaccines testing, bioanalytical issues, and life-cycle management of assays
    - Mr.Francis Dessy, Lead Clinical Assays Validation, GlaxoSmithKline
    • Multiple practical case studies on Vaccine Clinical Assay Development
      • Early method development
      • Approaches to Fit for Purpose (FFP) Validation ("Qualification")
      • Strategies for method troubleshooting
    • How to connect assay variability, critical reagents, bringing criteria and assay intended use?
    • Conversion of validated LBA to multiplex formats
      • What needs to be included in validation
      • Multiplexing strategies for multivalent vaccines
    • General Method Development strategies for LBA
      • Dos & don't for Vaccine method development
      • How to deal with lack of negative samples in assay development and validation?
      • How to demonstrate linearity over a 6-7 log broad range?
    • Novel approaches to bioanalysis of vaccines responses
      • Considerations in vaccine LBA with the use of novel delivery systems
      • Concomitant vaccines testing
    • Strategies for life-cycle management of assays
      • Role of assay standardization to progress in vaccine development and regulatory assay acceptance

  • Lesson 10
    Bioanalytical Performance of LBA for Vaccine: Clinical sample analysis, assay maintenance, bridging, trending, proficiency panel
    - Dr. Holger Koch, Global Head Clinical Serology & Assay Strategy, Takeda
    • Case studies on Clinical Sample Analysis
      • Control strategies for vaccine clinical assays
      • Assay performance
        • Maintenance
        • Bridging
        • Trending
        • Stability Monitoring
    • How to develop an assay stability monitoring program through the whole vaccine development program (Phase I-IV and in 10-20 years)
    • Bridging data from multiple studies
    • Development of proficiency panel for Vaccine Clinical Assays


What's new in Critical Reagents and Impact on Method Development?
  • Topic 11
    Updates & New Case Studies on Critical Reagents Generation, Characterization, Troubleshooting, and Long-Term Management: Building on WRIB recommendations and evolving trends
    - Dr. Nisha Palackal, Director Protein Biochemistry, Regeneron
    • Building on the 2019 White Paper in Bioanalysis Recommendations on Critical Reagents with novel practical case studies
    • Focus on Generation, Characterization and Maintenance
      • Updates for Critical Reagents discussion/training within the Bioanalytical Community and importance of learning from each other
      • Assessment of binding properties (e.g., cross-reactivity to matrix components), binding kinetics and any altered binding issues due to labeling or storage
    • Current Best Practices for critical reagent characterization, Troubleshooting, and Long-Term Management

  • Topic 12
    Focus on Bridging Methods for Critical Reagents: Case Studies
    - Dr. Christine Grimaldi, Head Biotherapeutic Bioanalysis Group,Boehringer Ingelheim
    • Influence of Critical Reagents Life Cycle Management on Assays for successful biotherapeutics discovery & development
      • Changing critical reagents and need to re-develop, re-validate and cross validate assays
    • Challenges and methods to bridge critical reagents
      • Use of lot bridging technique
      • Design of Lot bridging studies
        • Use of stability samples and freshly prepared QCs
        • Evaluation of meaningful changes
      • Ratios of the QCs' mean concentrations between old and new lots
      • Other bridging experiments that measure the effect of using different lots of critical reagents
    • Case studies on proper design of bridging methods for Critical Reagents

  • Topic 13
    State-of-the-art Strategies for Rapid Critical Reagent Screening and Optimization: What's new from last year WRIB recommendations?
    - Ms. Alison Joyce, Group Lead Discovery Bioanalytical and Critical Reagents Group Biomedicine Design, Pfizer
    • Reagent Pair Screening and Optimization for LBA
      • Screening reagent pairs
      • How to increase ruggedness on multiple platforms
    • Evaluation of optimal performance
      • Sensitivity
      • Range
      • Robustness
      • Reproducibility
    • Critical Reagents and major impact on assays performance
      • Strategic reagent screening and characterization to minimize assays failure
    • Case studies and lesson learnt from numerous biotherapeutic programs

  • Lesson 14
    Critical Reagent Strategies for Maintaining LBA Assays at CROs during Clinical Development: Practical case studies from pharma experience
    - Dr. Weili Yan, Scientific ManagerBioanalytical Sciences, Genentech
    • Critical Reagents management & expiration at CRO
      • No expiration until fail a retest
      • Evaluations to confirm key characteristics and assay performance
      • Harmonization on critical reagents expiration between Pharma & CRO
      • Overall process for Stability testing
    • Application of 2019 White Paper in Bioanalysis recommendations on critical reagents re-testing
    • Case studies on critical reagents approaches for maintaining LBA at CROs during clinical development
      • How to maintain an appropriate characterization from early discovery to post-marketing

Final Agenda Agenda at a Glance