8:00 am Check-In & Coffee
8:20 am Chair’s Opening Remarks
WORKING WITH INHERENT RESTRICTIONS TO ENABLE EFFECTIVE RESOURCE MANAGEMENT & MAXIMIZE DATA OUTPUT
8:30 am Innovative Solutions for Maximizing Data Output Despite Limited Gene Therapy Product Sample Sizes
Synopsis
• Exploring strategies to maximize inflammation output from limited sample sizes
• Leveraging orthogonal methods for physicochemical characterization to overcome limitations posed by small sample volumes
• Highlighting advances in small-volume technologies to enhance assay sensitivity and reduce material requirements
9:00 am Accelerating Analytical Capabilities to Increase Throughput & Deliver Quicker Results on Critical Quality Attributes
Synopsis
• Exploring how advanced analytical technologies facilitate rapid identification and evaluation of crucial CQAs
• Introducing techniques for automation of sample prep procedures
• Presenting data on novel automated immunoassays used to increase throughput and widen the dynamic range
9:30 am Speaking Slot Reserved for Bio-Techne
10:00 am Morning Refreshment Break
BIOASSAYS
Optimizing Cell-Based Bioassays for Enhanced Sensitivity Towards AAV
11:00 am Establishing Potency Strategy for AAV Gene Therapy Products
Synopsis
• Due to the complexity of Gene Therapy products, it is challenging to establish a one-size-fits-all strategy for potency method development
• Close communication with the research team is critical for defining potency method platforms in CMC
• We have developed strategic guidelines to define efficient approaches to potency matrix development for different AAV products
11:30 am Identifying & Selecting Specific Cell Lines to Optimize AAV Transduction Efficiency & Increase AAV Potency Assay Sensitivity
Synopsis
• Mechanisms for Screening Cell Lines: Exploring strategies to screen cell lines based on therapeutic area relevance to improve assay sensitivity
• Cell Line Optimization Towards AAV: Discussing methodologies for optimizing cell lines to enhance AAV transduction efficiency
• Comparing the use of stable versus transfected cell lines to minimizing batch-to-batch variability in potency assessment for rare disease
• Reducing the multiplicity of infections in cell-based bioassays to increase assay resolution and minimize cellular toxicity
PHYSICOCHEMICAL PROPERTIES
Enabling Extended Vector Characterization Across Delivery Platforms
11:00 am Developing a Novel CE-Based Method for the Assessment of Genome Integrity
Synopsis
• Optimizing an analytical strategy using the Sciex pa800 method to effectively characterize AAV genome integrity
• Comparing the relative amount of full and partial genome in stressed and non-stressed samples
• Demonstrating sensitivity of the method to detect loss of genome in different stressed conditions
11:30 am Innovative Mass Spectrometry Techniques for AAV Characterization in Gene Therapy
Synopsis
• Introduction to a novel mass spectrometry-based method for detailed analysis of Adeno-Associated Virus (AAV) structure and conformation
• Discussion on how this technique addresses previously unmet needs in the in-depth characterization of AAVs used as delivery vectors in gene therapies
• Insights from recent research findings and their implications for advancing gene therapy applications
PROCESS DEVELOPMENT & CMC
Enhancing Assay Automation & Method Development for CMC
11:00 am Leveraging Automation for High-Throughput Assay Development & Enhanced Method Comparability
Synopsis
• Discussing the benefits of automation in increasing throughput and efficiency in assay development and execution
• Presenting case studies of automated methods, including qPCR, HCP, and ELISA-based assays
• Exploring the complexities of automating assays, including difficulties in scripting, and establishing communication between wet lab scientists and automation engineers.
• Discussing thresholds for acceptable differences to establish comparability between automated and manual assays and ensure data reliability
11:30 am Developing an AAV9 Gene of Interest-Specific Expression Relative Potency Assay for CNS Indications
Synopsis
• Presenting a case study of assay development, including MOA-based potency assay design, optimization, and data analysis
• Generating pre-qualification data to establish acceptance criteria and analytical target profile
• Demonstrating successful method transferability to QC
12:00 pm Lunch & Networking
MOLECULAR BIOLOGY
Improving Capsid Identity & Extended Characterization
1:00 pm Evaluating Oxford Nanopore Technology for AAV & Plasmid Identity
Synopsis
• Introducing Oxford Nanopore technology for plasmid and AAV identity analysis to fast-track readouts
• Extended characterization of the viral payload
• Custom bioinformatics for data analysis versus epi2me
• Outlining the benefits of in-house NGS identity testing in a PD lab relative to outsourcing
1:30 pm Determining the Relevance of Next Generation Sequencing (NGS) as an Orthogonal Measure of Genome Integrity Analysis
Synopsis
• Using NGS to identify fragmented genomes and rebuild genome sequences
• Method development for understanding the genomic diversity of viral vectors
• Optimizing the sample preparation phase of NGS to reduce the number of false positives/false negatives and increase confidence in genome integrity analysis
2:00 pm Speaking Position Reserved for Qiagen
PHYSICOCHEMICAL PROPERTIES
Enabling Extended Vector Characterization Across Delivery Platforms
1:00 pm Consequences of Physical & Chemical Degradation on Product Quality & Determine Relevant CQAs
Synopsis
• Discussing the planning and design considerations for effective forced degradation studies, including linking exposure to heat, high pH, and other stress conditions to aggregation and potency readouts
• Addressing the stability indicating analytics required for forced degradation studies and exploring the lack of clear guidelines from regulatory agencies
• Examining the impact of physical and chemical degradation on product quality, including effects on capsid integrity and potency, to inform CQA assessment and process development mitigation strategies
1:30 pm Mass Spectrometry & Peptide Mapping Analysis of AAV Capsid Proteins & Characterization of Their PTMs
Synopsis
• Development of an in-house method for PTM analysis
• Leveraging a forced degradation case study to robustly characterize the importance of PTMs
2:00 pm Mastermind: Assessing the Impact of Post Translational Modifications on Viral Vector Activity to Inform Process Development & Formulation & Maximize Capsid Yield
Synopsis
• Determining the impact of deamidation, acetylation, and other relevant changes on viral vector entry
• Identifying biophysical markers to assess whether any PTMs can be considered CQAs for potency
• Linking post translational modifications to encapsulation efficiency to inform upstream manufacturing protocols, increase transduction efficiency and increase capsid yield
PROCESS DEVELOPMENT & CMC
Flexibility in Gene Therapy Manufacturing Processes & Supply Chains
1:00 pm Adapting to Challenges in Gene Therapy Manufacturing: Process Optimization & Ultra Filtration Enhancements
Synopsis
• Addressing low success rates in ultra filtration steps by adapting equipment and refining processes for small-scale gene therapy production
• Implementing strategies to improve yield and efficiency by minimizing sample numbers and refining manufacturing workflows
• Exploring techniques for maintaining regulatory compliance throughout the manufacturing and testing processes
1:30 pm Ensuring Manufacturability in Gene Therapy: Strategies for Commercial-Ready Processes
Synopsis
• Learning how to implement controlled bioreactors and chromatography in small-scale models to accurately reflect commercial manufacturing processes
• Understanding the importance of early-stage evaluations to determine yield, affordability, and quality, ensuring processes are ready for large-scale production
• Discovering strategies for identifying and resolving solubility problems and maintaining product quality throughout the manufacturing process
2:00 pm Optimizing Gene Therapy Supply Chains: Real-Time Batch Analysis
Synopsis
• Learning how connecting patient data with the manufacturing process can streamline operations and reduce inefficiencies, leading to faster and more reliable production cycles
• Discovering techniques for implementing real-time batch analysis to identify and mitigate potential issues during the production process, ensuring higher quality and compliance
• Understanding the challenges of managing large volumes of data across disconnected systems and exploring solutions for creating a centralized data repository to enhance the analysis and decision-making
2:30 pm Afternoon Refreshment Break
UNDERSTANDING THE IMPACT OF DIFFERENT ASSAY TYPES – CLOSING THE CIRCLE TO BUILD A ROBUST CMC PACKAGE
3:30 pm Working with Process Development Teams to Develop an Effective in Process Analytical Strategy & Inform Robust Manufacturing Qualities
Synopsis
• Using PAT tools to provide quick insights beyond release panel assays, enhance product robustness, and ensure reliability by identifying manufacturing issues promptly
• Working with process development teams to inform decision-making and optimize/adjust manufacturing conditions in real-time to ensure stable and robust gene therapy products
4:00 pm Physicochemical Properties & In Vitro Potency Impact of Distinct Empty Capsid Populations in Recombinant AAV Preparations
Synopsis
• Detection of distinct populations of empty capsids during anion exchange chromatography (AEX) purification of recombinant adenoassociated viruses (rAAVs)
• Comprehensive characterization of the structural and compositional diversity of these empty capsids • Assessment of the impact of empty capsid populations on the in vitro potency of purified rAAVs
4:40 pm Utilizing AUC, NGS & Viral Protein Data to Link Biophysical Characteristics to Vector Activity
Synopsis
• Using orthologous biophysical assays to predict potent fractions of a novel Bacovirus
• Discussing practical applications of this integrated approach to optimize vector design and selection and advance the development of more potent candidate molecules