Specialized Workshop Th1Thursday September 29, 2022: 7am - 5:30pm - Topic List

"LBA Novel Technologies, Novel Modalities, and Novel Challenges"

Innovative Tools/Instrumentation/Approaches; Advanced Strategies in Automation; Advancements in Critical Reagents Deep Functional Characterization; Problem Solving for Novel Modalities, Rare Matrices and Complex NAb Assays

(You can scroll down to see the details of each topic)

Part 1: Novel Technologies & Automation in LBA

  • Topic 1:

    Impact of Novel Technologies in Bioanalysis for advanced PK Assays and Biomarker Assays Applications
    Dr. Sally Fischer, Senior Director Bioanalytical Sciences, Genentech

  • Topic 2:

    Advanced Strategies in Automation of Large Molecule Bioanalysis
    Dr. Mark Ware, Scientific Director, Janssen

  • Topic 3:

    Automated Bioanalytical Workflow for PK & ADA LBA Development
    Dr. Linlin Luo, Director Immunogenicity & PK Assay Group, Merck

  • Topic 4:

    Automation or "industrialization" for sample pre-treatment/ADA enrichment for Immunogenicity assays
    Ms. Alison Joyce, Director, Discovery Bioanalytical & Critical Reagents, Pfizer

Part 2: Novel Modalities & Novel Method Development/Validation Challenges

  • Topic 5:

    Challenges in Enzyme Assays Development/Validation for Gene Therapy: How to overcome lacks enzyme reference material, to establish QC ranges and manage long term assay performance
    Dr. Yanmei Lu, Vice President Biomarker and BioAnalytical Sciences, Sangamo

  • Topic 6:

    Complex LBA Method Development/Fit for Purpose Validation for Cas9: From Critical Reagents selection to different molecule recognition, Step by step method development challenges & solutions
    Dr. Neil Henderson, Associate Director Nucleotide Bioanalysis Group, AstraZeneca

  • Topic 7:

    Challenges with Development/Validation of a PK assay to Measure a Variably Glycosylated Fusion Proteins
    Dr. Gizette Sperinde, Principal Scientist Bioanalytical Sciences, Genentech

Part 3: Novel Modalities & Rare Matrices

  • Topic 8:

    PK & ADA Assays for Drugs Administered through Nonstandard Routes (Intravitreal & inhaled): Is there benefit to analyze in these matrices?
    Ms. Kelly Coble, Director of Bioanalytical Sciences, Boehringer Ingelheim

  • Topic 9:

    Novel Strategies in Method Development/Validation and Sample Collection/Analysis for LBA Ophthalmology Bioanalysis: Special focus on Novel Modalities and Limitations in Sample Availability
    Dr. Kay-Gunnar Stubenrauch, Strategy Area Lead Genomic Medicine Bioanalytics & Biomarkers, Roche

Part 4: Problem Solving for Complex NAb Assays

  • Topic 10:

    A Brand-new Idea to Overcome Drug Interference in NAb Assay: “The PEG precipitation of the drug/NAb complex
    Dr. Weifeng Xu, Director, Merck

  • Topic 11:

    Novel Duplex Assay to Assess NAb Response to Bispecific Biotherapeutic with Weak Binding to T Cell Receptor
    Dr. Bonnie Wu, Associate Scientific Director Biologics Development Sciences, Janssen

Part 5: Critical Reagents Deep Characterization

  • Topic 12:

    Importance of Assay Reagent Binding Kinetics for Bioanalysis: What Factors Drives the Bioanalytical Assay Strategy and will we be able to Predict the "Best Assay Protocol"?
    Dr. Gregor Jordan, Lab Head Bioanalytics & Biomarkers, Roche

  • Topic 13:

    Importance to Develop of an Appropriate Target-Antigen Binding Assay to evaluate ADA Impact on Drug-to-Target Interaction
    Dr. Gregor Lotz, Group Leader Bioanalytics & Biomarkers, Roche

  • Topic 14:

    Evolved Applications of Biolayer Interferometry for Bioanalytical Assay Development
    Dr. Florian Neff, Lab Head Bioanalytics & Biomarkers, Roche

Part 5: White Paper in Bioanalysis

  • 2022 White Paper on LBA Topics:

    Consensus & Conclusions on LBA Topics for 2022 White Paper

 

Topic DETAILS of Th1

Part 1: Novel Technologies & Automation in LBA
  • Topic 1

    Impact of Novel Technologies in Bioanalysis for advanced PK Assays and Biomarker Assays Applications
    Dr. Sally Fischer, Senior Director Bioanalytical Sciences, Genentech

    • Novel Technologies and their huge potential in bioanalysis
      • Enabling more informed drug development decisions
      • Interrogation of drug and biomarkers in serum/plasma as surrogate for site of action
      • Evaluations at the site of action when needed
    • Multiplexing technologies
      • Quantification of multiple biomarkers for evaluation of complex diseases
    • Evaluation of PK and Biomarkers at lower volumes and concentrations
      • Novel technologies for nasal samples for asthma indications
      • PK assay for an oral antigen binding fragment of heavy chain only antibodies (VHH drug) at very low concentration in serum
      • Quantification of neuro biomarkers in serum and/or plasma instead of CSF to eliminate invasive spinal tap collections
      • Specific technologies for biomarkers quantification for ophthalmology indications in vitreous fluid
    • Case Studies: Illustration of the capabilities of promising Novel Technologies in bioanalytical field
  •  

  • Topic 2

    Advanced Strategies in Automation of Large Molecule Bioanalysis
    Dr. Mark Ware, Scientific Director, Janssen

    • Current developments in the fully automated processes in assay development and bioanalysis of biologics
      • When and how to apply automation
      • Application of automation highly suitable to large molecule bioanalysis
      • Degree of automation and standardized processes
    • Expectations of outcomes of automation
      • Benefit such investments and disruptions as to how operate
      • Infrastructure changes and workspace defined layouts
    • Impact upon workforce
      • Adaptation of laboratory scientists to this new environment
      • Right skill sets
      • Needs for IT infrastructure
    • Implementation, execution, and monitoring
      • Hurdles of the on-site testing before implementation
      • Computer systems validation (CSV)
      • Monitoring /maintenance to ensure long term benefits
      • Regulatory considerations
    • Case Studies: Integrating automated approaches in bioanalysis
  •  

  • Topic 3:

    Automated Bioanalytical Workflow for PK & ADA LBA Development
    Dr. Linlin Luo, Director Immunogenicity & PK Assay Group, Merck

    • Need for implementing Automation into LBA
      • Meeting the increasing demand for project support
    • Innovative workflows using automation
      • Implementation for both PK & ADA assay development
      • Comprehensive design of experiment (DOE)
      • Choosing the right Assay Format for robust PK assays
      • Using DOE and automation t to optimize the ADA assay conditions
    • Automation LBA improvements
      • Robustness
      • Reproducibility & Reliability
      • Throughput
    • Screening & Selecting Optimal Reagents
      • Quick ranking of candidate anti-IDs
      • Identification the best anti-ID pairing
    • Case Studies: Automated bioanalytical workflow consisting of automated assay implementation and data analysis

  • Topic 4:

    Automation or “industrialization” for sample pre-treatment/ADA enrichment for Immunogenicity assays
    Ms. Alison Joyce, Director, Discovery Bioanalytical & Critical Reagents, Pfizer

    • Need for efficient sample pre-treatment steps for ADA assays & NAb assays to address issues such as
      • Drug Tolerance
      • Matrix Interference
    • Multiple acid steps and drug-coated magnetic beads used on pre-treatment steps
    • Current problems in sample pre-treatment methods
      • Method Complexity
      • Time-consuming
      • Difficulty to transfer to CRO
    • Innovative use of automation as alternative methods to magnetic bead approach
      • Using tips pre-loaded with streptavidin resin for automated ADA enrichment
    • Case Studies: Automated pre-treatment/ADA enrichment for Immunogenicity assays

Part 2: Novel Modalities & Novel Method Development/Validation Challenges
  • Topic 5:

    Challenges in Enzyme Assays Development/Validation for Gene Therapy: How to overcome lacks enzyme reference material, to establish QC ranges and manage long term assay performance
    Dr. Yanmei Lu, Vice President Biomarker and BioAnalytical Sciences, Sangamo

    • Considerations on enzymes and inherited metabolic disorders
      • Therapeutic approaches for treating hereditary monogenic disorders with enzyme deficiency
    • Enzyme Assays vs LBA
      • Utility of enzyme activity readout during preclinical and clinical drug development
      • Advantage of using enzyme activity assay over protein level quantification technologies such ligand binding assays (LBA)
    • Enzyme Assay Technologies
      • Different enzyme assay technologies
      • Assay designs
      • Recording modes
    • Unique challenges of enzyme assay for Gene Therapy
      • Enzyme assay for Gene Therapy vs Enzyme Replacement Therapy (ERT)
    • Fluorometric enzyme assay Development/Validation
      • Reading 4-Methylumbelliferone (4-MU) product conversion for Gene Therapy Biomarker Analysis
      • Selection of instrument
      • Quantitation strategy of using 4-MU product curve instead of recombinant enzyme calibration curve
      • Assay development considerations and steps
      • Assay validation parameters
    • Case Studies: Recommended practice in enzyme assay development and validation to quantify the activity of transgene expressed enzyme in gene and cell therapy as a biomarker readout during preclinical and clinical drug development.

  • Topic 6:

    Complex LBA Method Development/Fit for Purpose Validation for Cas9: From Critical Reagents selection to different molecule recognition, Step by step method development challenges & solutions
    Dr. Neil Henderson, Associate Director Nucleotide Bioanalysis Group, AstraZeneca

    • Bioanalytical challenges in the quantification of Cas9 endonuclease bacterial protein
      • Cas9 technology characterization & bioanalytical assays
      • Efficacy assessment & Safety considerations
      • Specificity evaluation
      • Delivery to the target cells
    • LBA approaches to measuring CRISPR Cas9 RNP complex components
      • Method Development strategies
      • Historical and future landscape of Critical Reagent sourcing
      • Overcoming cell extract DNA interference
    • Need for different selective methods for
      • spCas9
      • SaCas9
    • Helpful information coming from bDNA for guide RNA
      • Guide RNA probe design
      • Dual guide delivery
      • Extraction efficiency
      • Cross reactivity with transfected DNA when trying to measure expressed RNA transcript
    • Case Studies: LBA Method Development/Fit for Purpose Validation for Cas9

  • Topic 7:

    Challenges with Development/Validation of a PK assay to Measure a Variably Glycosylated Fusion Proteins
    Dr. Gizette Sperinde, Principal Scientist Bioanalytical Sciences, Genentech

    • Protein Glycosylation for Large Molecule therapeutic drug candidates
      • Significant impact on PK
      • Difficulty in controlling Protein glycosylation during the manufacturing process of large molecules
    • Highlighting the challenges in quantification of a of highly glycosylated product
      • Sialic acid variability
      • Different assay development strategies
      • Ensuring robust and unbiased detection of the therapeutic candidate
    • Detection of variably glycosylated proteins
      • Impact from both critical reagents and platforms used in bioanalysis
      • Evaluation of assay platform and formats to ensure detectability of a wide set of sialic acid variants
    • Identification of the best assay platform able to meet requirements
      • Simoa assay
      • Hybrid assay
    • Case Studies: Development/Validation of a PK assay to Measure Variably Glycosylated Fusion Proteins

 

Part 3 Novel Modalities & Rare Matrices
  • Topic 8:

    PK & ADA Assays for Drugs Administered through Nonstandard Routes (Intravitreal & inhaled): Is there a benefit to analyze in these matrices?
    Ms. Kelly Coble, Director of Bioanalytical Sciences, Boehringer Ingelheim

    • Influence of Route of Administration on Systemic Exposure and Immunogenicity of biotherapeutics
      • What is known about absorption into systemic circulation after administration?
      • Biotherapeutic agents administered via a nonstandard routes
      • What are the gaps with ocular or inhaled administration of biotherapeutics?
    • Impact of the specific properties of the different tissue matrices considered for nonstandard routes of administration
    • Challenges for sample collection & sample processing of human tissue matrices
      • Bronchoalveolar lavage fluid (BALF)
      • Ocular tissues
      • Tumor tissues
    • Bioanalytical assay considerations for development of assays for non-standard tissue matrices
      • PK assays and ADA assays from matrices collected from nonstandard routes of administration
    • Case Studies: Bioanalysis of biotherapeutics administered via nonstandard routes and specific tissue bioanalysis

  • Topic 9:

    Novel Strategies in Method Development/Validation/Sample Collection/Analysis for LBA Ophthalmology Bioanalysis: Special focus on Novel Modalities and Limitations in Sample Availability
    Dr. Kay-Gunnar Stubenrauch, Strategy Area Lead Genomic Medicine Bioanalytics & Biomarkers, Roche

    • Bioanalytical Lesson Learned from VABYSMO (faricimab-svoa) for the treatment of adults with Neovascular (Wet) Age related Macular Degeneration (AMD) and Diabetic Macular Edema (DME)
    • Understanding how the molecular structure and route of administration (Intravitreal - IVT) of engineered bi-specific antibody drive the selection of appropriate assays for PK, ADA and Soluble Targets analysis in
      • Pre-clinical Development
      • Clinical Studies
    • Challenges, solutions, and learnings over a decade of method development, validation and sample collection/analysis for PK and ADA Assays
      • Overcoming the limitations in sample availability by using ultra-sensitive analysis
      • Drug Interference and Soluble Target Inference (against two multimeric soluble targets, VEGF and Ang2) for ADA testing with special respect to ophthalmology specific issues
    • Case Studies: Novel data on the relationship of the ADA response in different matrices (plasma vs ocular fluids) and use of Ultra-sensitive platforms to overcome sample availability

 

Part 4: Problem Solving for Complex NAb Assays
  • Topic 10:

    A Brand-new Idea to Overcome Drug Interference in NAb Assay: “The PEG precipitation of the drug/NAb complex
    Dr. Weifeng Xu, Director, Merck

    • Drug Interference is one of the biggest challenges for NAb assay
      • Major impact especially for mAb therapeutics in oncologic indication with high dose and long half-life
    • Considerations on the most popular sample pre-treatment used to mitigate drug interference
      • Acid dissociation followed by biotinylated-drug to extract NAb (BEAD)
      • Known impact of harsh acid which often denature acid sensitive NAb PC
      • Need for large amount of biotin-drug
      • Need for expensive SA-magnetic beads
      • Overall tedious and time-consuming process involving beads
    • Applications of a novel use of PEG to selectively precipitate drug/NAb complexes and wash away free drug
      • Very small amount of biotin-drug is added to dissociated drug and NAb
      • Much mild acid to form biotin-drug/NAb complex.
      • Complex binds to MSD plate and is detected with a sulfo-tagged drug target
    • Advantage of this PEG precipitation and acid dissociation with biotin-drug as assay drug (PABAD)
      • One step mild acid treatment instead of two-step harsh acid in BEAD;
      • Less than 1/1000 of biotin-drug needed
      • No magnetic beads needed
      • No additional assay drug needed
    • Case Studies: Novel approaches to overcome Drug Interference in NAb Assay

  • Topic 11:

    Novel Duplex Assay to Assess NAb Response to Bispecific Biotherapeutic with Weak Binding to T Cell Receptor
    Dr. Bonnie Wu, Associate Scientific Director Biologics Development Sciences, Janssen

    • Considerations on the Bioanalytical strategy for Bispecific Therapeutic Proteins
      • Customization of the Bioanalytical strategy
      • MOA and impact on assay selection
      • Cell-based vs Non-cell based assays
      • Target biology
      • Availability of the critical reagents
      • Positive Control (PC) used
    • NAb assay format for bispecific therapeutic proteins
      • Selection of Non-cell based assay format for assessment of NAb to bispecific therapeutic proteins
      • Challenge to develop NAb assays for bispecific MAb biotherapeutic that is engaged in T Cell redirected cancer cell killing but has weak binding to T cell receptor
    • Method development using an AlphaLisa technology platform
      • Optimization of duplex AlphaLisa assay
      • Strategies for a robust NAb detection
      • Assay sensitivity, robustness, and drug tolerance
      • Key assay parameters for the duplex AlphaLisa-based NAb assay
    • Case Studies: Development of a movel Duplex Assay to assess NAb response to Bispecific Biotherapeutic with Weak Binding to T Cell Receptor

 

Part 5: Critical Reagents Deep Characterization
  • Topic 12:

    Importance of Assay Reagent Binding Kinetics for Bioanalysis: What Factors Drives the Bioanalytical Assay Strategy and will we be able to Predict the "Best Assay Protocol"?
    Dr. Gregor Jordan, Lab Head Bioanalytics & Biomarkers, Roche

    • Modelling & Simulation Strategies:
      • What are the minimum parameters we need to select a platform as suitable?
      • Use of mathematical assay model to support the decision for when the soluble target could be bound by several soluble binding partners
    • Approaches for selecting an Assay Platform/Protocol
      • Relevant parameters for Free or Target Engagement analysis.
      • KD value, Rate Constants or Reagent Concentrations
      • Need to evaluate the KD value
    • Critical Reagents Selection
      • Has the Capture Reagent always to be less affine to the soluble target compared to the drug?
      • Do Critical Reagents (Capture and Detection) have to be in terms of binding properties comparable, and in case not, what can be done to optimize the assay protocol?
    • How to optimize assays protocols
      • Equilibrium Based or Kinetic Based
      • Considering a Total assay protocol:
      • Shall we skip Target Engagement(TE) assays and go for Total assay analysis and then calculate the Free portion?
    • Case Studies: Bioanalytical assay strategies to predict the most suitable assay platforms/protocols

  • Topic 13:

    Importance to Develop an Appropriate Target-antigen Binding Assay to evaluate ADA Impact on Drug-to-target Interaction
    Dr. Gregor Lotz, Group Leader Bioanalytics & Biomarkers, Roche

    • Importance to develop proper target-antigen assay Critical Reagents able to
      • Be a mimicry of the target antigen in patients to detect functional drugs
      • Assess ADA interference on target binding
    • Need for a deeper functional characterization of recombinant target-antigen critical reagents for clinical assays to use them as an adequate reagent tool for PK assay development
      • Biochemical binding
      • Biological activity
    • Patient-derived ADAs interference with the drug binding to recombinant and cell membrane target
      • Evaluation of adequate comparability of loss of drug binding function
    • Use of PK assay with the characterized recombinant target-reagent tool to show
      • Correlation of ADA response
      • Loss of exposure in clinical trial
    • Case Studies: Development challenges and strategies for Target-antigen Binding Assay Critical Reagents

  • Topic 14:

    Evolved Applications of Biolayer Interferometry for Bioanalytical Assay Development
    Dr. Florian Neff, Lab Head Bioanalytics & Biomarkers, Roche

    • Characterization of bioanalytical assay Critical Reagents:
      • Biolayer Interferometry (BLI) vs
      • Surface Plasmon Resonance (SPR)
    • Head-to-head evaluation to measure
      • Binding kinetics
      • Affinity of molecular interactions
    • Method comparison for quality determination of
      • Anti-idiotypic antibodies
      • Antibody conjugates
    • Use of Biolayer Interferometry to accelerate screening for optimized sample pre-treatment conditions
      • Evaluation of Drug/Target complex dissociation
    • High potential of Biolayer Interferometry for fast characterization of ADA responses
    • Case Studies: Recent advanced applications of Biolayer Interferometry (BLI) vs SPR for LBA Critical Reagents development in bioanalysis




Agenda at a Glance Agenda at a Glance