Specialized Workshop Th1 - Thursday Sep. 30, 2021: 7am - 5:30pm - Lecture List

"Conventional & Novel Critical Reagents: Recent Issues in Development and Management for Established/Emerging Technologies/Modalities"

Part 1: Novel Critical Reagent Modalities: "Thinking out of the Box"

Part 2: Advanced Approaches in Critical Reagents Selection

Part 3: Challenges in Critical Reagent & Positive Control Generation for ADA Assays

Part 4: Critical Reagent Management for Lot-to-Lot Bridging and Assay Comparability

 

Lecture DETAILS of Th1 – Conventional & Novel Critical Reagents

Part 1: Novel Critical Reagent Modalities: "Thinking out of the Box"
  • Lecture 1
    Impact of Restrictions on Animal-Derived Antibodies on Critical Reagents: Current Regulatory Status and Industry "Thinking out of the Box"
    - Dr. Boris Gorovits, Vice President Biomarker Discovery and Bioanalysis, Sana Biotechnologies
    • Non-animal-derived antibodies
      • Mature technology for critical reagents generation
      • Well-defined reagents equivalent to animal-derived antibodies
      • Better reagents with performance superior to animal-derived antibodies
      • Polypeptide sequences established as part of the generation process
      • Improved Reproducibility
      • Better supply sustainability of critical reagents
    • Thinking out of the box and alternative proposed solutions
      • Phage-display produced monoclonal antibodies
        • Current data on phage-display technique and its development as alternative antibody production
        • Potential issues with production of polyclonal antibody
        • Selection of antibodies using a universal recombinant library
      • DARPins (Designed Ankyrin Repeat Proteins)
        • Genetically engineered antibody mimeticnon-immuno globulinproteins
        • Derived from natural ankyrin repeat proteins
        • Highly specific and high-affinity target protein binding
        • Already successfully used and binders against several hundred targets have been created
        • Low-cost production in bacteria and the rapid generation of many target-specific reagents
      • Affimers
        • Small proteins that bind to target molecules with similar specificity and affinity to that of antibodies
        • Applications of non-antibody binding proteins to mimic the molecular recognition characteristics of monoclonal antibodies
      • Affibody
        • Small proteins engineered to bind to a large number of target proteins or peptides
        • Applications of these high affinity proteins and their ability to imitate monoclonal antibodies
    • Multiclonal non-animal-derived antibodies
      • Recent results showing exceeding performance vs monoclonal antibodies
      • Lower unspecific reactions and higher reproducibility vs polyclonal antibodies
    • Industry concern on the proposal by the European Commission to eliminate the use of antibodies derived from animals
      • Level of impact on LBA practice
      • Strong progress in reduction of animal use in the drug discovery and development process
      • Expected significant reduction of animal use in production of antibody reagents
      • Need to generate more data on non-animal derived antibodies
    • Case Studies: New ideas and methods, approaches currently tested. Use offully synthetic libraries and potential risk to produce lower affinity reagents
  •  

  • Lecture 2
    Novel Critical Reagent Modalities: Pfizer positive/negative experience with Aptamers, Peptides, Virus-Like Particles (VLPs) and other non-antibody reagents
    - Ms. Beth Leary, Principal Scientist Bio Medicine Design Group, Pfizer
    • Building on recent Industry/Regulators’ recommendations on Use of Aptamers versus Traditional Antibodies
    • Investigations on novel critical reagents modalities to demonstrate that non-antibody reagents can perform as substitutes for antibody reagents
      • Why consider alternatives to antibodies for ligand-binding assay (LBA) reagents?
    • Aptamers
      • Synthetic oligonucleotides that adopt a 3D structure and bind to target molecules
      • Major benefit of aptamers as synthetic reagents
      • Less lot-to-lot variability
      • No issues with loss of antibody producing cell lines
    • Pros and cons for using Aptamers routinely in LBA
      • Advantages of Aptamers, reagents versus traditional antibody approaches
      • Considerations to be taken into account for substitution of the IgG protein framework
      • Non-antibody reagents costs vs traditional antibodies
      • Availability and companies able to produce them
    • Use of Critical Reagents generated from a fully synthetic gene library
      • Protein scaffolds able to provide certain advantages in particular applications
      • New alternative binders introduced in the market
    • Case Studies: Direct comparison of antibody and non-antibody reagents to determine if performance is comparable and can be used in LBA
  •  

  • Lecture 3
    Recent Issues in Conventional & Novel Critical Reagents
    - Dr. Andrew Mayer, Group Leader Bioanalysis, GlaxoSmithKline
    • Building on recent Industry/Regulators’ recommendations on Critical Reagents
    • Issues in Conventional Critical Reagents
      • Variability between different batches produced in different animals at different times
      • Difficulty in reproducing a well-performing batch
      • Current approaches to minimized batch-to-batch variability
    • Potentially intrinsic lack/low of specificity
      • Considerations on the impact of additional light chains from various origins on specificity
    • Novel Reagents with improved affinity and specificity
      • Optimization of the binding kinetics
      • Directed evolution the association and dissociation rates
      • Ability to successfully fulfil the requirements of demanding applications as the natural immune response
      • Reduced cross-reactivity
      • Improved stability, affinity, avidity specificity and functionality
      • Multi-epitope recognition
    • Comparing the use of animal and non-animal derived antibodies as critical reagents
      • Impact on in bioanalytical assays
      • Non-animal derived critical reagents can be fit for purpose
      • Flexibility to use animal derived reagents when necessary.
    • Case Studies:Challenges & solutions in the application of conventional &novel critical reagents

 

Part 2: Advanced Approaches in Critical Reagents Selection
  • Lecture 4
    Advanced Approaches in Selection of Critical Reagents for Sensitive Clinical PK Assays
    - Ms. Kelly Coble, Director Bioanalytical Sciences, Boehringer Ingelheim
    • Building on recent Industry/Regulators’ recommendations on Using Multiple Platforms for Critical Reagents Selection
    • High Throughput Screening
      • Use of a fully automated system to perform high throughput screening of potential critical reagent pairs for use in early clinical PK method development
      • Evolution in process for assessment of reagent binding specificity and binding potency
        • Establishing blocking, Partial blocking andNnon-blocking reagents
    • Increasing ruggedness using Multiple Platforms
      • Use of high sensitivity platforms including SMCxPro, GyroLab and MSD during critical reagent evaluation and selection
      • Use of Octet and LBA assessment of reagent binding specificity
      • Use of Biacorefor binding potency assessment
      • Generation, screening and pairing of multiple anti-Ids
      • Risk of missing possible pair candidates
    • Evaluation of optimal performance
      • Sensitivity, Range, Robustness and Reproducibility
      • Critical Reagents and major impact on assays performance
      • Strategic reagent screening and characterization to minimize assays failure
      • Insights on strategies for selection of candidate pairs
    • Case Studies: Highlight approaches as well as some key lessons learned during the evaluation of Critical Reagents
  •  

  • Lecture 5
    Development of Novel Technologies to Enable Efficient Generations of High-Quality Critical Reagents: AstraZeneca’s experience
    - Dr. Ellen O’Connor, Associate Director Biopharmaceutical Development, AstraZeneca
    • Novel Technologies and applications in bioanalytical reagents characterization
      • Improvement of critical reagent controls with new orthogonal techniques
      • High-throughput kinetic information
      • Assay failures based on lack of in-depth biophysical characterization
      • Using automated platforms to make reagent generation more efficient and improve turn-around times
    • Application of Immobilized Metal Affinity Chromatography (IMAC) in critical reagent selection
      • Applications of IMAC to show how the isoelectric point of antibodies influence the conjugation reaction and formulation conditions
      • Affinity chromatography as preferred method to capture proteins from cell culture harvest
      • Highly selective ligand to bind a Fc-containing in monoclonal antibodies
    • State-of-the art strategies in Critical Reagent purification
      • Development of new technologies to enable efficient reagent generations
      • Strategic reagent screening and characterization to minimize assays failure
    • Case Studies: Lesson learned in critical reagent sciences from recent biotherapeutic programs and solution to generate high quality reagent to drive the R&D pipeline
  •  

  • Lecture 6
    Importance of Affinity Determination in Critical Reagent Development: Pfizer experience based on multiple case studies
    - Ms. Teresa Caiazzo, Senior Principal Scientist BioMedicine Design Group, Pfizer
    • Building on recent Industry/Regulators’ recommendations on Critical Reagents Affinity Determination
    • Current approaches for selection of high-affinity, stable and well-characterized Critical Reagents
      • Importance of well-characterized critical reagents as essential requirements for high performing ligand binding assays (LBA)
      • Identification and understanding of key characteristics of mAb reagents related to LBA performance to provide a strategic advantage to improved LBA performance
      • Issues with specific antidrug and/or anti-target Critical Reagents
      • Impact of Critical Reagents on the assay design and format
      • Critical Reagents cross reactivity
      • Affinity Determination by Biacore, Octet or similar applications
    • Assessment of binding properties
      • Critical Reagent Affinity and relation to bioanalytical assay function
      • Cross-reactivity to matrix components
      • Binding kinetics
      • Altered binding issues due to labeling or storage
    • Case Studies: Lesson learned on Importance of Affinity Determination from numerous biotherapeutic programs. Exploitation of the kinetic attributes of monoclonal antibody (mAb) reagents to facilitate fit-for-purpose applications in LBA and minimize development times
  •  

  • Lecture 7
    The Effect of Degree of Labeling (DoL) & Aggregation on Pharmacokinetic Ligand Binding Assays (PK LBA)
    - Dr. Shannon Chilewski, Principal Scientist, Bristol-Myers Squibb
    • Building on recent Industry/Regulators’ recommendations on Critical Reagents for PK LBA
    • Consideration on Labeling of Capture & Detection Reagents for PK LBA
      • Antibodies conjugation with a chemical moiety (Tag) for
        • Being coated onto the plate (capture)
        • Signal generation in the assay (detection)
    • Impact of Degree of Labeling (DOL) of Tags on the antibodies
      • Influence on stability and efficacy
      • Binding alteration of the physical and chemical properties of both antibody and conjugate
    • Issues with the Aggregation propensity of unconjugated antibody
      • Compounding or reduction during labeling
      • Affect reagent stability over time
    • Vital importance of effects DOL & Aggregation on different PK LBA to ensure high assay performance
      • Understanding the effects of DOL & Aggregation
      • Application of a systematic approach to evaluate different levels of DOL & Aggregation
      • Testing on three types of commonly used PK LBA
        • Chemiluminescent ELISA
        • MSD-ECL assay
        • Gyrolab assay
    • Case Studies: LDOL & Aggregation impact on PK LBA and determination of assay characteristics including an effect on binding, assay sensitivity, and assay range

 

Part 3: Challenges in Critical Reagent & Positive Control Generation for ADA Assays
  • Lecture 8
    Selection and Application of "Clinically Relevant" Positive Control (PC) anti-Ids for ADA Assays
    - Dr. Christine Grimaldi, Distinguished Research Fellow, Boehringer Ingelheim
    • Building on recent Industry/Regulators’ recommendation on Expectations for Approaches Regarding Positive Controls
    • Current challenges in the development of drug-related surrogate Positive Control
      • Anti-ID PC as a surrogate for ADA
      • Clinical significance/relevance of binding epitopes, binding potency and isotypes for patient ADA vs lab generated anti-ids
    • Screening paradigms to PC selection and impact on ADA assay method
      • Assessment of affinity/avidity and relevance to assay sensitivity
      • Neutralizing vs binding anti-id PCs
      • Selection of PCs for multidomain molecules
      • Impact of acid treatments on PCs
    • Impact of PC on ADA Assays performance
      • Assay performance using polyclonal vs monoclonal vs pooled monoclonal anti-ids
      • Impact of PC binding potency on assay sensitivity
    • Are improved Positive Controls representative of patients’ immune response?
      • Strategies to overcome the risk of developing ADA assays with high affinity PC that are not reflective of the patients’ immune response
    • Case Studies: Importance of selecting clinically relevant Positive Controls for improving the performance ofADA assays and of testing multiple PCs and assay conditions
  •  

  • Lecture 9
    Determination of Binding Affinity for Positive Controls (PC) and Critical Reagents for ADA Assays
    - Dr. Yang Xu, Senior Principal Scientist, Merck
    • Building on recent Industry/Regulators’ recommendations on Critical Reagent and Positive Control Characterization for ADA Assays
    • Improved approach in the development of PC
      • Specificity
      • Acid tolerability
      • mAb PC more consistent from lot-to-lot
      • pAb PC considered more representative
    • Importance of characterization of suitable Positive Controls
      • Best Positive Control for assay performance and determination of Assay Sensitivity
      • Altered Drug Tolerance in ADA assays due to Positive Control and Critical Reagents issues
      • Positive Control influence on Cut Point Determination
    • Novel strategies
      • Current challenges in the development of drug-related surrogate Positive Control
      • Well-characterized Positive Controls and ADA Assays performance
      • Purity and stability
    • Case Studies: Recent improvementsin the Generation & Characterization of Positive Controls (PCs) for ADA Assays
  •  

  • Lecture 10
    Reagent Troubleshooting for ADA Assay - Lesson learned from recent biotherapeutic programs
    - Dr. Hanjo Lim, Scientific Manager BioAnalytical Sciences, Genentech
    • Building on recent Industry/Regulators’ recommendations on Reagent Troubleshooting
    • Current strategies for conjugation and BPC capabilities
      • Streamlined reagent request and report systems/processes
    • ADA Assays - complete investigations to find the rootcause
      • Method Troubleshooting using characterization strategies
      • Follow‐up analytical investigation
      • Strategy to monitor the quality of ADA LBA critical reagents
    • Overall consideration on how to perform Bioanalytical troubleshooting of recent LBA impacted by the characteristic of the Critical Reagents
      • Step-by-step investigation
      • Tools for assuring reagent consistency for long‐term LBA performance and high‐quality data generated
      • Issues with conjugate incorporation ratios that affected assay performance
      • Issues with buffer component that affected conjugation efficiency
      • Troubleshooting binding affinity that affected assay performance
    • Case Studies: Lesson learned from recent biotherapeutic programs involving ADA assays. In depth investigations on the Impact of LBA Critical Reagent Quality on ADA Assay performance

 

Part 4: Critical Reagent Management for Lot-to-Lot Bridging and Assay Comparability
  • Lecture 11
    Vaccine LBA - Strategies for a successful Critical Reagent Management
    - Dr. Francis Dessy, Clinical Assays Science Advisor, GlaxoSmithKline
    • Building on recent Industry/Regulators’ recommendations on Critical Reagent Management
    • Influence of Critical Reagents Life Cycle Management on Vaccine LBA
    • How to minimize the risk of interruptions/delays to/of the supply chain of Critical Reagents for Vaccine Clinical Assays
      • Bad planning, forecasting and management
      • Unavailability of the right reagents at the right time
      • Delays for method development and validation
      • Impact on program decisions, time and resources
    • Common failures inCritical Reagents Life Cycle Management
      • Inefficient and insufficient implementation of adequate characterization technologies
      • Production of low-quality reagents
    • Strategies for new Lots of Critical Reagentsand need to
      • Re-develop
      • Re-validate
      • Cross-validate Vaccine LBA
    • Case Studies: Considerations to ensure a well-designed Critical Reagent Management for long term support of Vaccine LBA
  •  

  • Lecture 12
    Critical Reagent Management for Lot-to-Lot Bridging and Assay Comparability: Janssen’s Experience & Perspective
    - Dr. Mark Ware, Scientific Director, Janssen
    • Building on recent Industry/Regulators’ recommendations on Critical Reagent Lot-to-Lot Bridging
    • Consistency of Performance of Critical Reagents
      • Foundation of maintaining accurate and precise assay results of sample analysis throughout the lifespan of its use
      • Strategies to control production, labeling, storage and use of critical reagents
    • Considerations on lot-to-lot bridging
      • Lot-to-lot bridging cannot always be expected to be a replicate of current performance
      • Variability within acceptance criteria may be observed throughout the lifecycle of an assay
    • Variability of Performance of assays over time
      • Presence of variability also when additional control parameters are in place
      • Variability can exist across and within studies, with no determined singular root cause
      • Considerations impactful differences in sample accuracy or precision
      • Considerations on Trend Analysis over the years
    • Case Studies: Proper design ofbridging methods for Critical Reagents. Influence of Critical Reagents Life Cycle Management on LBA for successful biotherapeutics discovery & development




Agenda at a Glance Agenda at a Glance