Main Workshop DAY 3 - Thursday April 4, 2019, 7am to 6:45pm
"LBA, Cell-based Assays/Flow Cytometry & Emerging Technologies DAY"
(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
Novel Bioanalytical Developments in Cancer Immunotherapy & Gene Therapy
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Topic 1:
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Topic 2:
On-going developments in CAR-T Bioanalytical Strategies & Challenges by Flow Cytometry
Immunogenicity - Interpretation & Implementation of the 2019 FDA Final Immunogenicity Testing Guidance: Focus on Key Components of "Immunogenicity Testing of Therapeutic Protein Products - Developing and Validating Assays for Anti-Drug Antibody Detection Guidance"
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Topic 3:
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Topic 4:
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Topic 5a:
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Topic 5b:
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Topic 6:
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Note:
Due to the time limite in the Main Workshop Day, the following part of the 2019 FDA Final Guidance will be discussed during the F1 Full Day Training Session on Friday: Pre-Existing Antibodies, Cut-Point of Screening Assay Cut-Point of Confirmatory Assay, Cut-Point of Neutralization Assay, Cut-Point in the Target Population
Vaccine Assays & Biomarker Assays
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Topic 7:
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Topic 8a:
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Topic 8b:
Vaccine Clinical Assays - From Clinical assay to Immunogenicity: GSK experience
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Topic 9a:
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Topic 9b:
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Topic 10:
2019 White Paper on LBA, Cell-based Assays/Flow Cytometry & Emerging Technologies
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Consensus and Conclusions on LBA, Cell-based Assays/Flow Cytometry & Emerging Technologies
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Interpretation & Implementation of the 2019 FDA Final Immunogenicity Testing Guidance
Day 3B: ICH M10 BMV Draft Guideline - Ligand Binding Assays
Harmonized Topics among US FDA, EU EMA, Health Canada, Japan MHLW regulations
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Topics will be announced once ICH Draft Guideline is Published
Unresolved Issues and On-going Industry/Regulators’ Discussions
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Topics will be announced once ICH Draft Guideline is Published
Panel Discussion on ICH M10 BMV Draft Guideline - Ligand Binding Assays
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Industry Recommendations to ICH M10 Major Participating Regulatory Agencies
- Dr. Brian Booth (US FDA / EWG Regulatory Chair)
- Dr. Jan Welink (EU EMA / EWG Member)
- Dr. Akiko Ishii (Japan MHLW / EWG Rapporteur)
- Dr. Yoshiro Saito (Japan MHLW / EWG Member)
- Dr. Anna Edmison (Health Canada / EWG Member)
- Ms Thais Correa Rocha (Brazil ANVISA / EWG Member)
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
- Recent applications of Flow Cytometry in Cancer Immunotherapy
- 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
- On-going research and achievements in Chimeric Antigen Receptor T-cells (CAR-T)
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
- Acid dissociation: Does it do what we think it does?
- 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
- Improving Drug Tolerance in ADA/NAb assays continues to be a major concern for the Regulatory Agencies
- 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
- Recent Industry/Regulators’ Recommendations on Positive Controls
- 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
- Recent case studies on Clinically Meaningful Immunogenicity
- 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
- Expected specific challenges
- 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
- ADA monitoring in serum/plasma and in ocular matrices
- Original data from an ophthalmology clinical program
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
- What is your PK assay telling?
- 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
- Consideration on the high number of Biomarkers developed for projects in discovery stage – "the exploratory one"
- 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
- Exploratory biomarkers discovery/determination and how to progress them into a prognostic/diagnostic biomarker
- 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?
- Addressing biomarker LBA & Flow Cytometry Accuracy issues
- Current discussions on Absolute Accuracy within the Flow Cytometry Community
- Struggles to establish a meaningful approach for validating method accuracy
- Regulatory Statement: