Topic DETAILS of Th1

• 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
  
  

• 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

• 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

• 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

• 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