Main Workshop DAY 3 - Thursday April 12, 2018

"LBA/Cell-based Assays, LM Biomarkers Validation, and Immunogenicity
Challenges and Solutions"

Day 3 Discussion Topic List

(You can click on each blue topic title below to see details, or simply scroll down to see details)

Chair: Dr. Lauren Stevenson, Head Development Biomarkers & Bioanalytical Sciences, Biogen

PK Assays
Chair: Dr. Lauren Stevenson, Head Development Biomarkers & Bioanalytical Sciences, Biogen

Biomarker Assays
Chair: Dr. Renuka Pillutla, Executive Director, Bristol-Myers Squibb

Immunogenicity Assays
Chair: Dr. Sue Richards, Pres Scientific Fellow, Sanofi

LBA/Cell-based Assays, LM Biomarkers Validation, and Immunogenicity Panel Discussion & Consensus for 2018 White Paper in Bioanalysis
Chair: Dr. Lauren Stevenson, Dr. Renuka Pillutla and Sue Richards

  • Panelists Industry: Dr. Patty Siguenza, Dr. Steve Keller, Dr. Yongchang Qiu, Dr. Heather Myler, Dr. Shobha Purushothama, Dr. Sally Fischer, Dr. Rafiqul Islam, Dr. Paul Rhyne, Dr. Virginia Litwin, Dr. Catherine Aversa Fleener, Dr. Yongliang Sun, Dr. Ian Catlett, Dr. John Kamerud, Dr. Chris Beaver, Dr. Roland Staack, Dr. Cherie Green, Dr. Meina Liang, Dr. Vellalore Kakkanaiah, Dr. Eginhard Schick, Dr. Priya Sriraman, Dr. Martin Ullmann, Dr. Judy Shih, Dr. Tong-yuan Yang, Dr. Gizette Sperinde, Dr. Joe Marini, Dr. John Smeraglia, Dr. Robert Kubiak, Dr. Rong Zeng, Dr. Giane Sumner, Dr. George Gunn, Dr. Shan Chung, Dr. Patrick Liu, Dr. Szilard Kamondi, Dr. Vibha Jawa, Dr. Daniel Baltrukonis, Dr. Seema Kumar, Dr. Kay-Gunnar Stubenrauch, Dr. Jonathan Haulenbeek, Dr. Nisha Palackal, Dr. Boris Gorovits, Dr. Rachel Palmer, Dr. Steven Piccoli, Dr. Lakshmi Amaravadi, Dr. Devangi Mehta, Dr. Becky Schweighardt, Dr. Albert Torri, Dr. An Song

  • Panelists Regulators: Dr. Daniela Verthelyi, Dr. Susan Kirshner, Dr. Yow-Ming Wang, Dr. Steve Bowen, Dr. Haoheng Yan, Dr. João Pedras-Vasconcelos, Dr. Christopher Leptak, Dr. Shashi Amur, Dr. Sean Kassim, Dr. Sam Haidar, Dr. Seongeun (Julia) Cho, Dr. Kimberly Maxfield - TBC, Dr. Meenu Wadhwa, Dr. Isabelle Cludts, Dr. Pekka Kurki, Dr. Shirley Hopper, Dr. Jan Welink, Dr. Elana Cherry, Dr. Akiko Ishii-Watabe, Dr. Yoshiro Saito



Discussion Topic DETAILS of Day 3

Chair: Dr. Lauren Stevenson, Head Development Biomarkers & Bioanalytical Sciences, Biogen

  • Topic 1
    PK, PD, ADA Data Correlation and NAb Assays Merit
    - Dr. Lauren Stevenson, Head Development Biomarkers & Bioanalytical Sciences, Biogen
    • Focus on impact of NAb assays results vs. PK, PD, ADA data correlation
      • Is it possible to make the same conclusions from this correlation without NAb data?
      • How much immunogenicity assessment is needed?
        • How much is too much?
        • When no additional value is gained from immunogenicity assessment?
    • Experience in interacting with US FDA to demonstrate the non-need for NAb assays
      • What are the key information needed in this discussion?
      • What do you need to demonstrate? Designing the framework
    • Strategies to identify the neutralizing effect without NAb assays
      • Lessons learnt from the development of multiple biotheraputics on the incidence and impact of NAb
      • Clinical value of NAb assays for low risk molecules
      • Further developments on the concept of “Biomarkers of NAbs” as an alternative approach based on the 2017 White Paper Part 3 Recommendations
      • When should NAb assays really be carried out and why?
    • Case Studies: Evaluation of PK, PD, ADA data correlation for multiple biotherapeutics and actual cases where NAb assays are not needed

  • Topic 2
    PK, PD, ADA Data Correlation and Current Experience with New Modality Biotherapeutics for Ophthalmology
    - Dr. Kay-Gunnar Stubenrauch, Senior Scientist Bioanalytics, Roche
    • Original data from an ophthalmology clinical program
      • Expected specific challenges
        • Small volumes
        • Complex sample collection
        • Rare ocular matrices
    • New complications
      • New Modality Biotherapeutics (NMB) validation for PK, ADA and soluble targets/biomarkers assays
      • Novel bioanalytical issues and possible solution
      • Unique challenges of immunogenicity testing of ophthalmology biotherapeutics
        • Highly concentrated drug into the eye
        • Direct drug administration into the eye
        • Long duration of biologics
        • Major drug interference
    • PK, PD, ADA Data Correlation
      • ADA monitoring in serum/plasma and in ocular matrices
        • Interpretation of drug exposure
        • Correlation with histopathology and Tox findings
      • PD Assays
        • Protein biomarkers quantification in ophthalmology at very low concentration and in small volume of aqueous humor
        • Use of Ultra-sensitive platforms
      • PK Assays
        • Challenges with ocular drug administration far from the site of action within the eye
        • Bioanalysis in the various ocular tissues & fluids for PK, formulation or device suitability
    • Case Studies: Share the most recent experience in PK, PD, ADA Data Correlation for New Modality Biotherapeutics for Ophthalmology

  • Topic 3
    Integrated Summaries of Immunogenicity
    - Dr. João Pedras-Vasconcelos, Immunogenicity Reviewer, US FDA
    • Regulatory recommendations from revised US FDA Guidance for presenting immunogenicity data including PK, PD, ADA Data Correlation
      • Experience gained during US FDA review of the Integrated Summary of Immunogenicity
      • Addressing potential issues when generating the integrated summary
        • Findings in dossiers
        • Most common missing information
        • Most common confusing/unclear information
      • How to present relevant information in the submitted dossier
      • How to minimize regulatory questions & potential concerns
    • How the Integrated Summaries of Immunogenicity can help to locate all the key information in a single place
      • PK, PD, ADA Data Correlation usefulness for assessing immunogenicity risk
      • Intrinsic immunogenic potential of the molecule
      • Product quality
      • Product administration
      • Dose regimen
      • Correlation of bioanalytical and clinical data
    • Case Studies: Impact on PK&PD efficacy in reporting immunogenicity results. Importance of multi-disciplinary information for Immunogenicity risk assessment


PK Assays
Chair: Dr. Lauren Stevenson, Head Development Biomarkers & Bioanalytical Sciences, Biogen

  • Topic 4
    Critical Reagent Characterization: What’s meaningful?
    - Dr. Jonathan Haulenbeek, Senior Research Investigator, Bristol-Myers Squibb
    • Most current recommendations on reagent characterization & stability studies
      • Experiments needed to ensure that critical reagents are stable over the course of use
        • How extensive should be the characterization of the stability of critical reagents, especially labeled reagents?
      • How to reassure the Regulators that the Industry has now better critical reagent control
    • Major impact of formulation/storage/purity of critical reagents on assays performance
      • Current industry standards on essential tests to ensure critical reagents characterization
      • Novel applications of biophysical characterization of critical reagents biotherapeutics
      • Pros & Cons in qualifying critical reagents by assay performance
        • Assay failures due to critical reagent
        • Ensuring biophysical similarity
        • Use of innovative orthogonal techniques
        • Importance of in-depth characterization of critical reagents at the early stages of method development
          • Issues in characterizing critical reagent in later stages
          • What is the best approach for exploratory studies
    • Case Studies: Ensuring critical reagents quality, stability, reliability and continuity throughout drug discovery & development to avoid Regulatory concerns

  • Topic 5
    Processes for Efficiently Handling Critical Reagents: Impact of critical reagents formulation changes on false positivity of ADA assays
    - Dr. Nisha Palackal, Associate Director Protein Biochemistry, Regeneron
    • Recommendations for establishing a comprehensive & robust program to handle critical reagents
      • Advanced strategies for generating
      • Characterizing critical reagents
      • Maintaining critical reagents over the life time of assays
    • Close collaboration between Bioanalytical & Critical Reagents Groups in establishing an effective program
    • Issues associated to critical reagents integrity for ADA assays
      • Understanding how changing the formulation of a critical reagent in an ADA assay can have a major impact on false positivity during clinical studies
      • Impact of integrity of critical reagents in overall ADA assay performance and generation of reliable immunogenicity data
        • Sever effect of aggregates present critical reagents
        • Strategies to ensure consistent long-term performance of ADA assays
        • What are the specific issues associated with labeled assay reagents?
    • Case Studies: Handling critical reagents and their impact on ADA assays

  • Topic 6
    Viral Vector-based GeneTherapy: What are the regulatory expectation for PK, PD and ADA Assays vs. Industry current approaches?
    - Dr. Boris Gorovits, Senior Director, Pfizer
    • The latest insights & regulatory developments on this highly attractive and fast evolving technique
      • Focus on what bioanalysis should measure for assessing drug exposure, responses and immunogenicity
    • All the measurements to ensure that
      • The gene of interest reached the target cells or tissues
      • Efficiency of transfection in the targeted cells or tissues
      • Pre-treatment measurement of antibodies as immunogenicity assessment
      • Pre-existing antibodies to Viral vector and possible impact on safety and efficacy
      • Relevance for post treatment Immunogenicity measurements of over-expressed proteins
      • Is immunogenicity on transfected cells really needed?
        • Continuing the discussion and reaching consensus from the 2017 White paper Part 3 Recommendation
        • Are vector ADA, anti-transgene and collect a baseline PBMC sample for ELISPOT enough?
        • Should cellular responses to the virus by clearing of transfected cells considered?
        • What if the transgene is expressed on a cell surface and in a non-immune privileged area?
    • Challenges addressed & unanswered concerns
      • Specificity & tissue targeting
      • Specific tissue/cell tropism
      • Selection of viral vectors
      • Immunogenicity issues
      • Understanding pharmacology & safety
      • Regulatory expectations
    • Case Studies: Status, ongoing challenges and regulatory expectation of viral vector-based gene therapy

  • Topic 7
    Current Bioanalytical Challenges in Gene Therapy and Possible Practical Solutions
    - Dr. Rachel Palmer, Lab Operational Director, Sanofi
    • How bioanalysts are dealing with the increasing number of Gene Therapies contains both vector and transgene that are in clinical development
    • Actual bioanalytical data to support
      • PK behavior
      • Systemic & tissue exposure?
        • Vector
        • Transgene
      • Vector Shedding
        • Excretion/ secretion of the viral vector outside the body
      • Immunogenicity
    • Bioanalytical challenges when administration is at the disease site
      • Importance of bridging systemic and tissue exposure
    • BMV and fit-for-purpose (FFT) validation for methods used for Gene Therapies in support of Clinical Studies
      • PK assay for Viral Vector concentrations (RT-qPCR)
      • PD assay for transgene expression (LBA)
      • ADA assay for overall antibody testing (LBA)
    • RT-qPCR application in Regulated Bioanalysis
      • Biodistribution
      • Vector Shedding
      • Should RT-qPCR become a standard instrument in bioanalytical laboratory?
      • Recommendations for RT-qPCR validation criteria
    • Case Studies: Bioanalytical methods to support Gene Therapy Clinical Program: method development, validation and sample analysis challenges


Biomarker Assays
Chair: Dr. Renuka Pillutla, Executive Director, Bristol-Myers Squibb

  • Topic 8
    US FDA Perspective on Biomarker Qualification and the Importance of Assay Performance
    - Dr. Christopher Leptak, Director OND Regulatory Science Program/Director Biomarker Qualification Program, US FDA
    • Regulatory Input on Day 2B & Panel Discussion on Day 3

  • Topic 9
    EU EMA Perspective on Biomarkers Assay Development & Validation in Europe: “Concept Paper on Predictive Biomarker-Based Assay Development in the context of drug development and lifecycle”
    - Dr. Shirley Hopper, Medical Assessor, UK MHRA
    • Regulatory Input on Day 2B & Panel Discussion on Day 3

  • Topic 10
    Reaching a Biomarker Community Consensus on Disagreement Points of the Duke-Margolis/C-Path/US-FDA White Paper: Evaluation of Supporting Recent Protein Biomarkers Actual Case Studies & Regulatory Perspective
    - Dr. Shashi Amur, Scientific Advisor, US FDA
    - Dr. Steven Piccoli, CEO, Neoteric Consulting
    • Recent advancements in the discussion around biomarkers qualification & validation
      • FFP (Fit for Purpose) & COU (Context of Use)
    • Focus on the analytical factors where consensus was not reached
      • Evaluation of new case studies
      • Impact of these key factors on Biomarkers Assay
        • Robustness
        • Reliability
    • Why was not consensus reached?
      • Scientific and regulatory expectations for BAV
      • Critical unresolved issues on biomarker assay performance
    • Understanding of limitations of industry best practices for method development and BAV
      • Relative Quantitative assays
      • Validation of Commercial Kits
        • Reagent and Kit lot variability
      • Very High/Low endogenous levels
      • Best practise for assessing specificity
      • Protein Biomarker Assay Calibrator Material
    • Harmonizing what discussed in the Duke-Margolis/C-Path/US-FDA White Paper with European and Japanese Agencies
    • Case Studies: Supporting data & evolving discussions for the finalization of the “draft framework outlining key criteria and best practices for biomarker assay performance expectations and validation” developed by US FDA, C-Path and Duke-Margolis Center in 2017

  • Topic 11
    Level of Validation Required for Biomarkers which are NOT part of Duke-Margolis/C-Path/US-FDA White Paper
    - Dr. Lakshmi Amaravadi, Global Head Bioanalytical & Biomarker Development, Shire
    • Current industry standards and updated recommendations on BAV not used for regulatory decisions
      • Discovery biomarker translation from pre-clinical into human clinical studies
      • Biomarkers for exploratory/investigational studies
      • Human biomarkers for PK/PD decisions
    • Fit-for-purpose (FFP) BAV
      • What’s new?
      • Status & application of the recommendations:
        • 2016 White Paper Part 3
        • Evolving thinking/practices
      • Challenges to implementation of FFP BAV
        • Pitfalls and solutions
    • How to navigate the complex environments for biomarker support?
      • Considerations to different biomarker types
      • What are the limits and considerations on using bioanalytical lab to support biomarker studies in all the stages of drug development?
      • Generating reliable biomarker data to support drug programs
        • Exploratory biomarkers & intended use of the biomarker data
    • Case Studies: Application of a Biomarker Assay as suited for context of use (fit-for-purpose)

  • Topic 12
    Current Challenges with Cell-Based Biomarkers Assays: Flow Cytometry validation, sample handling and data analysis
    - Dr. Devangi Mehta, Associate Director Biomarker and Bioanalytical Sciences, Biogen
    • Targeting Cell Surface Receptors as PD Markers
      • Strategies for the development of Cell-based Biomarkers Assays
    • Flow Cytometry Advanced Applications
      • Increased interest in Flow Cytometry within Bioanalytical Labs
      • Method development & FFP validation of robust methods
      • Method qualification
      • Recommendations implementation
        • Next step for the 2016 White Paper Part 3
    • Challenges & Solutions
      • Panel assays set up for more than 14-color
      • Minimizing cell biological variability and samples instability
      • Single-cell multiparameter analysis
      • Staining of peripheral blood samples
    • Flow Cytometry innovations & implementations
    • Case Studies: Fast development of Flow Cytometry as bioanalytical tool for Cell Surface Receptors PD Markers


Immunogenicity Assays
Chair: Dr. Sue Richards, Pres Scientific Fellow, Sanofi

  • Topic 13
    ADA Incidence & Clinical Relevance: Generating Clinically Relevant ADA Data - Industry & Regulatory Perspectives
    - Dr. Sue Richards, Pres Scientific Fellow, Sanofi
    - Dr. Yow-Ming Wang, Associate Director of Therapeutic Biologics Program, US FDA
    • Focus on ADA incidence impacting PK & Efficacy which is
      • Clinically relevant
      • Clinically meaningful over time (persistent responses)
        • MAbs
        • New types of Fc-derived constructs
      • Clinical relevance of ADA and the issue of drug interference in patient management
        • What have been the successful approaches for regulatory submissions when the ADA assay is affected by large concentration of circulating drug
    • Generating Clinically Relevant Immunogenicity Data
      • Giving a correct drug dose and drug exposure to patients
      • Setting Appropriate Assay Cut Points
        • An even more in-depth discussion continues in the Specialized Session F1 with case studies from numerous companies
      • Categorizing ADA Positivity
      • Modifying the outlier removal process
      • Reporting only persistent ADA
    • Better predictors of clinical impact
      • Should there be more focus on clinical relevance of ADA and not just incidence rates?
    • Case Studies: Determination of the relationship between ADA assay data and the Clinical Relevance with impact on PK and efficacy

  • Topic 14
    Understanding of Clinical Relevance of NAb to better understand the correct development of NAb Assays: Focus on the risks associated with ADA-mediated neutralization
    - Dr. Becky Schweighardt, Executive Director, BioMarin
    • What to consider to develop a clinically relevant NAb assay that measures an appropriate target
      • NAb assay data and how these data impacted patients safety and clinical interpretation
      • What are the risks associated with ADA-mediated neutralization?
        • Neutralizing antibodies against therapeutic proteins can potentially impact patient safety and directly mediate loss of drug efficacy
      • What are we measuring in-vitro?
      • Are NA assay results corroborated by results of another parameter?
    • Transient vs Persistent responses
      • Possible differential impact
    • What is the justification for multiple Nab Assays?
    • Case Studies: Usefulness of NAb assays data to provide safety/efficacy insights that impacted patient treatment decisions

  • Topic 15
    What is the meaningful Drug Tolerance Limit that should be targeted? What is the Sensitivity that Needs to be reached in “Drug-tolerant” ADA/NAb assays?
    - Dr. Albert Torri, Executive Director Bioanalytical Sciences, Regeneron
    • Current challenges in the correct management of Drug & Target Interference in ADA/NAb assays
      • Drug & Target tolerant NAb assays
        • What is the best approach to develop them?
        • What are the requirements?
        • What’s new (2016 & 2017 White paper Part 3 Recommendations) in the strategies and procedures to reduce interference in ADA assays?
      • Regulatory expectations for Drug Tolerance Limit in ADA/NAb assays
      • What concentration of the positive control should Drug Tolerance Limit be established?
      • How far should we push methodology to increase Drug Tolerance Limit that may also negatively impact detection of ADA?
        • What is the balance and how do we justify it?
        • What should we do if we cannot meet the desired Drug Tolerance Limit?
    • Impact of pushing for more sensitive and drug tolerant ADA assays
      • More sensitive and drug tolerant ADA assays may detect clinically irrelevant signals which may mask more relevant ADA responses
      • Are the more stringent assay requirements resulting in more “unevaluable” results being reported?
      • Is this clinically useful?
    • Case Studies: Considerations on more sensitive and drug tolerant assays able to detect clinically irrelevant signals which may mask more clinically relevant ADA responses

  • Topic 16
    Quality of ADA Assays & Standardization of ADA Assays - Regulatory Perspective
    - Dr. Meenu Wadhwa, Section Leader, UK MHRA-NIBSC
    - Dr. Pekka Kurki, Research professor, Finland Fimea
    • Considerations on the gap in mutual understanding between Regulators & Industry with the quality of data generated by ADA Assays
      • Are current ADA Assays good enough to find adverse effect on PK, Safety and Efficacy?
      • Can we improve the quality of current ADA assays through the Standardization of the Positive Control (PC)?
        • Generation of reference standards for immunogenicity assays
        • Using standardization of PC for setting targets for relative ADA assay performance
      • Improving generation and interpreting ADA data by Standardization of PC
        • Comparability of ADA assays by using of same reference standards PC in determining assay performance
        • Avoiding the risks associated with changing PC during ADA assay evolution
    • Overall goal: Identifying Clinically Meaningful ADA Incidence
      • Is Industry “afraid” to find ADA?
      • Or is it concerned about reporting of ADA incidence that goes on labels if highly sensitive assays result in high incidence but without clinical consequence?
      • Industry’s worry: “Missing clinical correlations with sensitive assays”
      • Regulators’ worry: “Insensitive assays may miss clinical correlations”
    • This fruitful discussion is aimed to bring both Industry/Regulators to work together to align on a better understanding and consensus on what constitutes Clinically Meaningful ADA Incidence and the ADA incidence that should go on the label
      • Building on 2017 White Paper Part 3 Recommendations
        • More focus on an analysis where ADA-positives are stratified according to the titer/concentration of ADA
    • Case Studies: Regulatory direct experience on ADA assay quality & performance and their ability to generate clinically meaningful ADA data

  • Topic 17
    Isotyping & Domain/Epitope mapping of ADA: Utility (or lack of) of ADA isotyping assays?
    - Dr. An Song, Sr Director, Genentech
    • Current risk-based strategy on when this may or may not be needed
      • Pros/Cons based on lessons learnt
      • The last step of the multi-tiered approach: screening -> confirmatory-> titer assessment -> neutralizing drug activity -> isotyping & domain/epitope mapping
    • Specific multiplexing technologies to identify isotypes and for domain mapping
      • Novel Applications
      • New Technologies
    • Importance of isotyping for defining IgE-mediated hypersensitivity
      • Development challenges for an IgE isotype ADA assay
      • Uso of isotyping to discriminate non IgE-mediated anaphylaxis
    • Are they early predictors of impact of NAb development?
    • Regulatory requests for isotyping
      • Why is Isotyping & Domain/Epitope mapping requested?
      • How often?
      • Removal of recommendation to screen for IgA and IgE testing unless there is a specific reason
      • What are the clinical needs?
    • Case Studies: Specific experience with IgE assays, IgG subclassing, domain/epitope mapping and interaction with Regulatory Agencies



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