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

Day 3A: LBA, Cell-based Assays/Flow Cytometry & Emerging Technologies

Day 3B: ICH M10 BMV Draft Guideline - Ligand Binding Assays

Day 3A Details:
LBA, Cell-based Assays/Flow Cytometry & Emerging Technologies

Novel Bioanalytical Developments in Cancer Immunotherapy & Gene Therapy

• Topic 1
  Using Flow Cytometry for Predicting Response to Immunotherapy: Case Studies of Strengths and Gaps in Clinical Validation
  • Recent applications of Flow Cytometry in Cancer Immunotherapy
    • Baseline and/or on-treatment frequencies of immune cell types measured by Flow Cytometry
    • Evaluation of significant correlation with response to cancer immunotherapy
  • Case studies & keys findings
    • Assessment of strengths and gaps in the Flow Cytometry assays developed and validated for this purpose
    • Challenges, strategies and considerations in Clinical Validation

 

• Topic 2
  On-going developments in CAR-T Bioanalytical Strategies & Challenges by Flow Cytometry
  • On-going research and achievements in Chimeric Antigen Receptor T-cells (CAR-T)
    • Flow Cytometry as a key technology in developing CAR-T personalized therapy
    • Patient’s T cells engineered to express artificial T-cell receptors
    • Possibility of surface expression of several antigens with the generation of CAR T-cells with dual specificity
    • Automated analyses of Flow Cytometry data for prediction of clinical response to CAR-T Cell Therapies
  • Scientific and regulatory challenges in development of CAR-T Therapy
  • Lesson learnt from the characterization of CAR-T Therapy
    • Evaluation of Flow Cytometry based approaches
    • Monitoring of cell products for use in Clinical Studies
  • CAR-T Therapy What’s Next?
    • How Bioanalysis can help CAR-T development?
    • Bioanalytical strategy, assay development and validation
    • Advanced use of Flow Cytometry bioanalytical methods in cell-based pharmacokinetics or cellular kinetics in adoptive cell therapy

Immunogenicity

• Topic 3
  What’s New for Improving ADA Assay Drug Tolerance & Sensitivity? Do Acid Dissociation, SPEAD, ACE, PandA, BEAD & Co do what we think they do? Is there any other alternative/innovative/less complex and time-consuming approach?
  • Improving Drug Tolerance in ADA/NAb assays continues to be a major concern for the Regulatory Agencies
    • What’s new from the 2018 White Paper Recommendations Part 3
    • What are the most recent progresses?
  • Pros and Cons of current approaches for improving Drug Tolerance
    • Acid dissociation: Does it do what we think it does?
      • What are the alternatives to the current Acid-Dissociation techniques?
    • Is there room for improvement to avoid sample pre-treatments issues?
      • Impact on sensitivity, precision, binding affinity, dimerization, denaturation, analyte loss
    • Are we losing ADA in the sample pre-treatments so that samples are determined as negative or low positive when actually ADA are lost in the process?
      • Can recovery test help avoid potential bias?
    • Is the magnetic beads technology able to overcome the issues of the other approaches?
      • Evaluation of the high specificity to remove therapeutic drug from study samples
  • Considerations on how Drug tolerance test is currently performed
    • Risk in over or under-estimating drug tolerance due to the use of surrogate Positive Control

 

• Topic 4
  Surrogate Positive Controls (PC) Challenges, Concerns, Appropriateness for ADA Assays: How to deal with recovery, purity, stability, multiple epitope recognition, HPC & LPC criteria and FDA draft Guidance
  • Recent Industry/Regulators’ Recommendations on Positive Controls
    • Practical application of the 2017-2018 White Paper in Bioanalysis Part 3
  • Strategies for the generation of feasible Positive Control for ADA Assays
  • Current challenges in the development of drug-related surrogate Positive Control
    • Novel antibody engineering approaches
  • Impact on ADA Assays performance
    • Multiple epitope recognition
    • Purity and stability
    • Positive Control influence on Cut Point Determination
    • Are improved Positive Controls representative of patients’ immune response?
    • Impact of PC on determining sensitivity
    • Strategies to overcome the risk of developing ADA assays with high affinity PC that are not reflective of the patients’ immune response
    • Experiment needed when the Positive Control is changed
  • Value/relevance of performing
    • “Recovery” of surrogate Positive Control in individual samples for ADA assay robustness
    • Stabilities in surrogate Positive Control for actual ADA sample stability
      • Best practices in the stability testing of ADA vs Positive Control
  • Implication of Recommended LPC failure rate in the new FDA Guidance
    • LPC levels for screening and confirmatory assays
  • Critical evaluation of actual Clinical ADA data in case of LPC failure rate higher than recommended from the new FDA Guidance

 

• Topic 5a
  Clinical Relevance of ADA and Impact on Bioanalysis - Better Understanding of Clinical Relevance of ADA: Are Highly Sensitive & Drug Tolerant ADA Assays Helping with Clinical Correlation or just Increasing the Noise? Building on 2018 White Paper Recommendations
  • Recent case studies on Clinically Meaningful Immunogenicity
    • Is the cost in ADA assay improvements in sensitivity providing clinical insights?
    • Decision-making immunogenicity data and analyses
    • Recent considerations in redefining Clinical Immunogenicity Assessment
    • What have we learned from using drug tolerant ADA assays in clinical setting?
    • What’s the clinical relevance of low affinity ADA?
  • Clinically Meaningful Immunogenicity Profile Impact on Safety & Efficacy
    • Essential ADA/Nab assays developed to characterize the immunogenicity profile
    • Interpretation of ADA data and determination of clinical impact
  • Updated data and lesson learnt with highly sensitive and drug tolerant immunogenicity assays in understanding the clinical impact of ADA
    • Focus on clinical relevance of ADA and not just incidence rates
    • Clinically relevant ADA data generated by
      • Modifying the outlier removal process used to generate ADA assay CP
      • Reporting only persistent ADA

 

• Topic 6
  PK, PD, ADA Data Correlation, Clinical Relevance and Current Industry Experience with New Modality Biotherapeutics for Ophthalmology
  • 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

PK & Biomarker Assays

• Topic 7
  Soluble Target and ADA Impact on PK Assays: Analyzing, Interpreting and Reporting Factors that Impact PK data
  • What is your PK assay telling?
    • Interpretation Factors that impact PK measurements of biotherapeutics
    • How presence of high concentrations of circulating target, assay reagents, format and conditions can significantly impact the PK parameters/profiles
  • Case studies on impact based on what drug species is being detected
    • Impact of soluble target on PK profiles
  • Understanding the PK assay as well as the strengths and limitations of the resulting data
    • Crucial data interpretation to draw accurate conclusions
    • Interpreting potential association of PK assays with clinical outcomes
  • Case studies where an outcome was impacted by misleading interpretation of PK Assays due to soluble target and ADA impact
    • How are ADA interference in PK assays being analyzed and reported?
    • Recommendations on How PK assays can advise on ADA
    • Type of outcome and general analysis description
    • Findings and interpretation

 

• Topic 8a
  Vaccine Clinical Assays - Challenges in Bioanalytical Method Development & Validation, Concomitant Vaccines Testing, and Life-Cycle Management of Assays: Merck experience
  • The assay development and validation process for vaccine at Merck
  • Bioanalytical challenges with vaccines
    • Concomitant vaccines testing
    • Bioanalytical methods
    • Life-cycle management of assays
    • Validation & troubleshooting of vaccine test
  • Case studies on Cell-mediated immunity assays
    • Flow Cytometry Assays
    • ELISPOT Assays
  • Applied vaccine clinical assay practices
    • Clinical efficacy endpoint in vaccine clinical trials
  • Regulatory expectations for vaccines bioanalysis
    • US FDA Center for Biologics Evaluation and Research (CBER) for Vaccine and Gene Therapy
    • EU EMA scientific guidelines on vaccines and Clinical evaluation of new vaccines
      • 2018 revised guideline on the clinical evaluation of vaccines

 

• Topic 8b
  Vaccine Clinical Assays - From Clinical assay to Immunogenicity: GSK experience
  • The assay development and validation process for vaccine at GSK
  • Case studies on humoral immunity assays
    • LBA clinical Immunogenicity assay
    • Main challenges in the vaccine field
    • How to establish assay reference standards
    • Determination control samples and their use and utility in determining assay acceptance
    • Antibody functional assays
      • Viral neutralization assays
  • Updates from the 2018 White Paper in Bioanalysis discussion on Cut Points for Vaccines
  • Statistical methods are used to
    • Determine the limit of blank (true negative samples)
    • Assay LOD and LLOQ
    • Most important features for Vaccine ADA assays

 

• Topic 9a
  Exploratory Biomarkers – What are they? Guidance on how to approach Exploratory Biomarker Assays for Internal Decision-making: What is meant by COU and FFP
  • Consideration on the high number of Biomarkers developed for projects in discovery stage – "the exploratory one"
    • How are "exploratory" biomarkers defined?
    • What are the criteria behind that definition?
  • Lack of industry focus on Exploratory Biomarkers vs Confirmatory/Regulated Biomarkers
    • Lack of guidance on how to approach biomarker assays for internal decision-making
    • Lack of comfort to decide internally what is Fit for Purpose (FFP) or exactly the Context Of Use (COU)
      • What is meant by COU and FFP for exploratory biomarkers?
    • Debunking the notion that FFP means not validated
      • "FFP is not the F-word of the biomarker world"
  • Working together to draft recommendation able to provide clarity within the Industry for exploratory biomarkers
    • Case studies on based on a specific biomarker of great interest across industry
    • Neurofilament light
      • A case study in COU iterative assay development for biomarkers

 

• Topic 9b
  Exploratory Biomarkers – What are they? Current Industry Standards to Handle Assay Development, Validation, Transfer and Refinement of Exploratory Biomarkers from Discovery into Development
  • Exploratory biomarkers discovery/determination and how to progress them into a prognostic/diagnostic biomarker
    • Current industry practice on how companies handle the transfer/refinement/qualification of methods from Discovery into Development for exploratory biomarkers
    • Is it possible to use biomarkers assays developed for Discovery in FIH clinical studies for an “exploratory biomarker”? If not, why?
    • How to transfer biomarker platforms from discovery to clinical studies?
      • Transferring a biomarker exploratory Hybrid LBA/LCMS assay to a "regulators’ preferred" traditional LBA for a confirmatory clinical biomarker
    • When does an "exploratory biomarker" become a "clinical biomarker"?
    • When does a "clinical biomarker" become a "diagnostic"?
  • Exploratory Endpoints in clinical studies
    • On-going debate on how to approach this issue
    • How to deal with possible but unconfirmed biomarkers
    • What are the regulatory expectations for exploratory biomarker assay used in clinical trials?
      • Characterization, data reporting or non-reporting

 

• Topic 10
  Accuracy in Biomarkers Assays? Establishing a Meaningful Approach to Determine Absolute Accuracy in LBA & Flow Cytometry Assays
  • Regulatory Statement:
    • "Accuracy is one of the utmost fundamental requirements for validation of any assay including fit-for-purpose biomarker assays"
    • How the industry can work together to overcome the lack of absolute accuracy for biomarkers assays not having an appropriate or exact reference standard
      • Currently, LBA & Flow Cytometry assays are able to measure only relative accuracy instead of absolute accuracy
    • From 2018 White Paper in Bioanalysis Part 3: "As flow cytometry assays lack the availability of reference standards and the data generally fall into the category of quasi-quantitative, it is thus not possible to validate accuracy in the traditional manner"
  • Establishing analytic acceptance criteria for quasi-quantitative assays for Biomarkers
    • Addressing biomarker LBA & Flow Cytometry Accuracy issues
      • Exploration within the different technologies
    • How to improve the current reference standards used in LBA and Flow Cytometry Biomarkers assays?
      • Need to find more appropriate reference material
    • Importance to carefully and reliable assess the differences between the endogenous protein and the reference material
      • Is it possible to quantify the impact on absolute accuracy of current reference standards vs endogenous biomarkers?
  • Current discussions on Absolute Accuracy within the Flow Cytometry Community
    • Struggles to establish a meaningful approach for validating method accuracy