Specialized Workshop T2 - Discussion Topic List

"Innovation in Flow Cytometry, Mass Cytometry, and Imaging Cytometry to Solve Complex Method Development Issues in Drug Discovery/Development and Clinical Applications: What’s New for Bioanalytical Labs using Cytometry for Biomarkers, Immunogenicity, Cell Therapy and Cellular and Immuno-Oncology Assays?"

Learn from advanced case studies on Fluorescence-Based Flow Cytometry reduction in the complexity, increase in automation & sensitivity and capacity to detect rare populations. Learn the major advantages & challenges in using Mass Cytometry (CyTOF) & Imaging Cytometry vs traditional Flow Cytometry. How can your assay be improved by extending Flow Cytometry capabilities to high-throughput / high-content, single-cell assays, unmet antibody specificities in a single sample and without spillover between channels?

  • Dr. Alessandra Vitaliti,Executive DirectorMolecular and Cellular Biomarker, Novartis
  • Dr. Lindsay King, Assoc. Research Fellow BioMedicines Design, Pfizer

New Insights in Automated Gating

Innovation in High Dimensional / High Parameter Flow Cytometry

Imaging Cytometry and Implementation of Complex Assays in Clinical Trials

Advantages & Challenges in using Mass Cytometry (CyTOF) vs traditional Flow Cytometry

White Paper on Cytometry

  • White Paper

    Consensus, Recommendations, and Panel Discussions on Cytometry
    - Dr. Lindsay King, Assoc. Research Fellow BioMedicines Design, Pfizer
    - and Cytometry White Paper Working Experts Group


Topic DETAILS of T2 – Flow Cytometry, Mass Cytometry, and Imaging Cytometry

  • Dr. Alessandra Vitaliti, Executive DirectorMolecular and Cellular Biomarker, Novartis
  • Dr. Lindsay King, Assoc. Research Fellow BioMedicines Design, Pfizer
New Insights in Automated Gating
  • Topic 1
    High Complex Flow Cytometry in Clinical Trials: Recent Developments and Innovations
    - Dr. Michael Hedrick, Director Clinical Flow Cytometry, Bristol-Myers Squibb
    • Ongoing need for implementation of high complex Flow Cytometric assays in Clinical Trials for
      • Pharmacodynamics
      • Mechanism of Action (MoA)
    • Flow Cytometry applications in longitudinal patient monitoring setting
      • Improvement of efficiency for
        • Bioanalytical assay validation
        • Clinical sample analysis
    • Novel use of automated approaches to analyze Flow Cytometry data from clinical trials
      • Generation of high quality and reproducible data
      • Reduction of manual analysis in complex Flow Cytometry studies
      • Automated analysis methods
        • High throughput
        • Efficiency
        • Consistency
        • Reproducibility
    • Case studies on analysis of large sets of Flow Cytometry data from Clinical Trials

  • Topic 2
    Advancements in Automated Gating Strategies Implementation for Quality Control and Sample Analysis: Case studies, lesson learnt and future trends
    - Mr. Alberto Hidalgo Robert, Scientific Researcher OMNI Biomarker Development, Genentech
    • Traditional manual data analysis in Flow Cytometry
      • Manual gating strategies
        • Current Best practice
        • Risk associated with subjective human-driven decisions
        • Possibility of introducing significant analytical variability
        • Inconsistency and lack of data integrity
    • Advanced automated data analysis in Flow Cytometry
      • Automated gating strategies
      • Feasibility of automated clustering algorithms to
        • Identity low quality samples
        • Perform sample clean up
        • Measure dimly expressed checkpoint marker
    • Evaluation of automated gating for
      • Standardized and reproducible tool for analyzing flow cytometry data
      • Comparison manual vs automated data analysis
      • Pros and cons of manual vs. automated analysis
      • Validation of automated gating
        • Evaluation of QC data generated from automated gating
    • Case studies on automated gating vs manual gating for high throughput and reproducible flow cytometry data analysis

  • Topic 3
    Clinical Applications Utilizing High Dimensional Flow Cytometry and Fit-for-purpose (FFP) Algorithms: Automated biased and unbiased analysis tools for Flow Cytometry
    - Dr. Shabnam Tangri, Chief Scientific Officer, Navigate/Novartis
    • On-going technological progress in Flow Cytometry
      • Entering in the era of high dimensional datasets at a single cell levels
    • Issues in analyzing high dimensional datasets by classical manual gating techniques
      • Impact of high subjective component
      • Massive amount of data remains unused
    • Need for novel computational algorithms for high dimensional Flow Cytometry
      • Unbiased analysis.
        • Automated biased and unbiased analysis tools for flow cytometry
      • Availability of new commercial platforms
      • New workflow to perform analysis of high-parameter flow datasets
      • Strategies to interrogate high-parameter data in a flexible manner
    • Identification of predictive markers of CAR-T therapies by High Dimensional Flow Cytometry
    • Case studies on clinical applications utilizing high dimensional flow cytometry and fit-for-purpose algorithms


Innovation in High Dimensional / High Parameter Flow Cytometry
  • Topic 4
    High Parameter Flow Cytometry: Instrument Harmonization and Panel Design
    - Dr. Sandra Nuti, Head Flow Cytometry Core Facility, Vaccines GlaxoSmithKline
    • Current Flow Cytometry applications & accessibility of High parameter Flow Cytometry
      • Continual growth of the biopharmaceutical industry
      • Fundamental role in understanding
        • Impact of biological drugs on the immune system
        • Examining immunological changes to determine drug-related effects at an individual cell level
        • Enabling the rapid collection of data related to multiple parameters from a heterogeneous cell mixture
        • Key technique employed to rapidly profile large populations of cells
    • Current shift to ultra-high parameter flow cytometry (20+ colors)
      • Impact of minor spectral discrepancies to produce false positives and false negatives
      • Need for additional fluorochromes
    • Instrument Harmonization and Panel Design
      • Development of high-parameter flow cytometry
      • Use of commercial software packages to facilitate
        • Panel design
        • Data acquisition
        • Data analysis
    • Case studies on high parameter flow cytometry instrument harmonization and panel design

  • Topic 5
    Current Challenges to Perform a >30-color Flow Cytometry Experiment: Limitation of available fluorochromes vs state-of-the-art flow cytometers able for multiple the simultaneous measurement
    - Ms. Megan McCausland, Translation R&D Scientist, Q2 Solutions
    • Applications of high dimensional multicolor Flow Cytometry
      • Comprehensive immunophenotyping
      • Quick and effective monitoring of immune status
      • State-of-the-art Flow Cytometers
        • Simultaneous measurement of up to 50 different parameters
    • Challenges to perform a >30-color Flow Cytometry experiment
      • Limitation of available fluorochromes
    • Protocols strategies
      • Traditional Separation of one sample into several panels of 8 to 10 colors
        • Issues with limited samples volumes
      • Novel parallel design
        • Advantages to conduct comprehensive analysis also with limited amount samples
    • Case studies on validated protocol to perform high dimensional multicolor Flow Cytometry

  • Topic 6
    Recent Developments in Multi-parameter Flow Cytometry and Assay Development Strategies
    - Dr. Vilma Decman, Director and Head Cellular Biomarkers, GlaxoSmithKline
    • Novel case studies on Multi-parameter Flow Cytometry
      • Applications in drug research, development and clinic
      • Examination of multiple targets on various cell subsets
        • Possibility to analyze limited sample size
      • Identification and enumeration of multiple cellular markers
      • Tracking of cellular markers across time and treatment
    • Multi-parameter Flow Cytometry in Immunotherapy
      • Development and validation of multi flow cytometry panels
      • Advanced assays in support of clinical trials
      • Real-time basis samples analysis
    • Efficient and reliable identification of therapeutic drugs PD

  • Topic 7
    Advanced Cytometric Biomarker Analysis in Clinical Development: Applications of 20+ color immune monitoring panel in Flow Cytometry
    - Dr. Richard Wnek, Scientific Director Translational Molecular Biomarkers, Merck
    • Overall growing need for advanced Flow Cytometric biomarker analysis
      • Clinical development
      • Immune oncology
    • Advanced strategies for comprehensive immune monitoring
      • Collection of peripheral blood
        • Clinical biomarker applications
        • Longitudinal patient monitoring
    • Limitations of traditional/conventional 10-18 fluorescent parameter Flow Cytometry
      • Use of multiple panels in the same patient sample tube
    • Strategies for developing a 20+ Panel Design
      • Assay optimization
        • Reagent
        • Sample resource
        • Processing efficiency
      • Innovative panel design
      • Instrument setting optimization
    • Possibility to enable application of 35+ color panels for immunophenotyping
    • Case studies on highly granular immunophenotyping of functional and interrelated cell subset interactions


Imaging Cytometry and Implementation of Complex Assays in Clinical Trials
  • Topic 8
    Recent Development in Imaging Cytometry
    - Dr. Alessandra Vitaliti, Executive DirectorMolecular and Cellular Biomarker, Novartis
    • Limitations of traditional Flow Cytometry
      • Low sample stability
      • Need of mid-high sample volumes
      • Not real images of
        • Analyzed cells
        • Spacial distribution of a marker within a cell or cell surface
        • Cellular interactions
    • Recent evolution in Imaging Cytometry
      • Combining single cell imaging with multi-parameter high-throughput single cell analysis
      • Imagining technologies
      • Electronics
      • Digital computing
    • Image cytometry
      • High-throughput microscopy and laser scanning cytometry
      • Image Flow Cytometry
      • Image Mass Cytometry
      • Discussion on advantages and disadvantages of different technologies
    • Case studies on advances in image cytometry providing complementary information to conventional flow cytometry

  • Topic 9
    High Parameter Flow Cytometry in the Clinic: Investigating Tumor Infiltrating Lymphocytes
    - Dr. Priscila Teixeira, Scientific Leader Clinical Protein Biomarker Development, Roche
    • Recent applications of High Parameter Flow Cytometry in the Clinic
      • Advanced platform for single cell analysis
        • Cellular complexity
        • Biological and genetic studies of immune cell development
        • In depth study of immune-mediated disease
    • Importance of Tumor Infiltrating Lymphocytes (TILs)
      • Association of TILs with cancer clinical outcomes
      • Characterization of subtype-specific immune-tumor spatial interactions
    • Predictive biomarker for cancer immunotherapy
      • Development of a high parameter Flow Cytometry panel to matched tumor and adjacent normal tissue samples
      • Flow Cytometric Analysis of intraepithelial lymphocyte content within solid tumors
    • Developed a robust method for application of high parametric Flow Cytometry in clinical settings
      • Instruments performance optimization
      • Panel design complexity
      • Harmonization of multiple instruments sharing the same configuration

  • Topic 10
    Recent Advancements in the Implementation of Highly Sophisticated Flow Cytometry Assays in Clinical Trials
    - Dr. Ulrike Sommer, Lab Head Cellular Biomarker, Novartis
    • Role of complex Flow Cytometry in Clinical Trials
      • Advancing therapies through novel Flow Cytometry Biomarkers
    • Discussion on case studies and lesson learnt for highly sophisticated Flow Cytometry Assays to demonstrated clinical endpoints in Clinical Trials
      • Focus on the importance of standardization and harmonization of Flow Cytometry assays for multi-center global clinical studies
      • How to ensure quality of data by ensuring assay robustness and reproducibility
    • Challenge in Flow Cytometry BAV in a global contest
      • Application of a robust FFP Validation protocol for Flow Cytometry assay to support clinical efficacy
      • Strategies with method transfer to clinical site and importance of on-site trainings


Advantages & Challenges in using Mass Cytometry (CyTOF) vs traditional Flow Cytometry
  • Topic 11
    Advancements in Mass Cytometry: How to Ensure Robustness and Reproducibility When Using Mass Cytometry for Clinical Research
    - Mr. Jose Estevam, Scientific Director and Center of Biomarker Innovation and Development, Takeda
    • Challenges with Fluorescent Flow Cytometry
      • Limitation to a few cell subsets
      • Technical constraints when combining multiple markers in a single tube
    • Mass Cytometry
      • Possibility to examine of up to 50 parameters
      • Unique use of heavy metal isotopes rather than fluorescent agents
      • Avoiding the interference of spectral overlap of fluorescent agents
    • Multiplexed staining
      • In depth characterization of cell populations
      • Identification within a single tube of
        • Increased number of functionally distinct cell subsets
        • Rare immune cell subsets
    • An unbiased evaluation of Mass Cytometry use in Clinical Research
      • How to Ensure Mass Cytometry robustness and reproducibility for applications involving samples from Clinical Trials
      • Limitation of Mass Cytometry
      • Evolution of Mass Cytometry technology
    • Case studies on application of Mass Cytometry in clinical development

  • Topic 12
    Application of Mass Cytometry for Preclinical BioTox programs: Combining high dimensional phenotyping and quantitative biomeasures
    - Dr. Lindsay King, Assoc. Research Fellow BioMedicines Design, Pfizer
    • Current Application of Mass Cytometry as an advanced tool for drug discovery
      • Deep Phenotyping by using high dimensional Mass Cytometry
      • Enrichment of Activation and Proliferation Markers by Mass Cytometry
    • Novel and powerful approach by Mass Cytometry
      • Interrogation of complex immunophenotypes
    • Increasing the number biomeasurements in preclinical settings
      • Enabling translational modeling
      • Possibility to perform quantitiative biomeasures
    • Mass Cytometry application for Metabolic and Immune Profiling
      • Profiling of immune cell phenotypes from circulation to tissue
      • Development and validation of high parameter Mass Cytometry
    • Case studies on application of Mass Cytometry in preclinical setting

  • Topic 13
    A Comprehensive Strategy for Development and Validation of Immunoncology-focused Mass Cytometry Panels
    - Mr. Gregory Hopkins, Sr. Assoc. Scientist Cellular Analytics Team, Bluebird Bio
    • Strategy for development and validation of Immunotherapy-focused Mass Cytometry panels
      • Specific Panel design
      • Extensive panel validation
      • High number of parameters assessed
      • Thorough optimization of the staining
      • Impacts of signal spillover
        • Measure the total spillover for each channel
        • Assessment of the complete spillover profile of each individual antibody
      • Establishment of expression profile of markers that are difficult to stain
    • Case studies on advanced use of Mass Cytometry in CAR-T Immunotherapy
      • Exploring clinically relevant attributes of autologous cells in CAR-T Immunotherapy
      • Correlating relevant attributes to T-cell phenotypes
      • Deep phenotyping of CAR-T and CD34 enriched drug products
    • Discussion on lack a comprehensive guideline on Mass Cytometry on how to design, validate, and test mass cytometry panels