7:50 am Chair’s Opening Remarks

EXPLORING NEW TECHNOLOGIES & ANALYTICAL APPLICATIONS IN GENE THERAPY DRUG DEVELOPMENT

8:00 am Optimization of the AAV expression and purification using dedicated HPLC system

  • Ales Strancar Managing Director, BIA Separations, part of Sartorius group

Synopsis

Adeno associated virus (AAV) is the leading vector in the field of gene therapy because of its low toxicity, good overall safety profile, and ability to maintain stable expression for long periods of time. It is therefore crucial to develop a robust and high efficiency platform for its manufacturing.

One of the key challenges in manufacturing viral vectors is to increase the ratio between empty and full capsids. The most efficient way is to design the USP to result in less than 10% of empty capsids. This can be realised by process optimisation using at-line HPLC to allow for analysis of the full and empty capsids ratio during the virus expression.

Fast and reliable in HPLC methods to allow for process optimisation USP and DSP, and assessing the purity of the final product using PATfix system will be presented.

8:30 am Methods for DNA Characterization in AAV

Synopsis

  • Optimal analytical methods to characterize DNA presence
  • Characterizing residual and encapsidated DNA size in AAV products

9:00 am How TEM based image analysis can reveal process impact on critical quality attributes

Synopsis

  • GMP-certified services using transmission electron microscopy (TEM) and image analysis for characterization of gene therapy products
  • Using TEM to identify structural parameters correlated to important biological functions for quality control, efficacy and safety
  • CryoTEM: robust and validated method for AAV full/empty/intermediate characterization

9:30 am Morning Refreshments & Speed Networking

BIOASSAYS

Track Chair: Reena Patel, Scientific Integrator, Janssen

DEVELOPING AN EFFECTIVE ANALYTICAL POTENCY ASSAY STRATEGY

10:30 am PANEL: Overcoming Hurdles to Potency Assay Changes from Early Through to Late-Stage Clinical Development

  • Lyndi Rice Head of QC Viral Vector, BioMarin
  • Yan Zhi Director Analytical Development, Spirovant Sciences

Synopsis

This panel will set out to discuss:

  • The challenges of developing a potency assay strategy.
  • Developing a potency assay strategy in line with regulatory expectations
  • What communication with regulatory bodies looks like when changing your potency assay
  • Best practice for potency strategy development

11:00 am Long Term Program Management for Potency Assays

Synopsis

  • Importance of long term potency management
  • Challenges of potency assay management
  • Communication with regulatory bodies in regard to potency assay management

11:30 am Quality attributes and extended characterization of gene vectors using light scattering

Synopsis

• Quantifying quality attributes of vectors is of increasing
importance as the gene therapy products enter into the
late development stages and QC environment.

• Batch dynamic light scattering (DLS) is for rapid screening,
size exclusion chromatography coupled to multi-angle
light scattering (SEC-MALS) for multiple AAV CQAs
quantification, and field-flow fractionation (FFF) with
MALS for quantifying AAV aggregates and extended
characterization of large gene vectors.

MOLECULAR BIOLOGY

Track Chair: Mimmi Ballard, Group Lead Cell Based Assays, Sana Biotechnology

UNCOVERING THE LATEST NGS APPLICATIONS

10:30 am Development and qualification of a NGS assay for control of vector genome integrity of AAV9 drug products

  • Huimin Helen Tao Senior Scientist, Analytical Development, Novartis Gene Therapies

Synopsis

  • Next Gen Sequencing Assay Applications
  • Vector Genome Integrity of AAV9, challenges and opportunities

11:00 am Characterization of capsid species and genome in rAAV products

  • Wei Zhang Associate Director, Analytical Development, Ultragenyx

Synopsis

  • AAV empty capsids obtained by physical or chromotographic separation
  • Smaller sized genome packaged inside AAV empty capsids
  • Smaller sized genome is identified by NGS

11:30 am Development Of Gene Of Interest (GOI) Expression Bioassay to Measure Relative Potency of a Gene Therapy Product

  • Savita Nair Senior Scientist Bioassay Development , Sangamo Therapeutics

Synopsis

  • Design and development of a quantitative RT-PCR assay to
    measure the early endogenous expression of target gene
    for product characterization studies

ANALYTICAL CHEMISTRY

Track Chair: TBC

THE AGE-OLD CHALLENGE: OPTIMIZING EMPTYFULL CHARACTERISATION

10:30 am Analytical Tools for Characterization of Empty/ Full/Partial Capsids

  • Santosh Khatwani Associate Director, Analytical Development, Sangamo Therapeutics

Synopsis

  • Methods for Empty/Full AAV capsid characterization
  • Sangamo’s approach
  • Orthogonal approaches
  • Mass spectrometry in E/F measurement

11:00 am Comparison of orthogonal methods of determining the relative abundance of empty and full AAV capsids

Synopsis

  • The relative abundance of empty and full AAV capsids in gene therapy vector preparations is a critical quality attribute
  • A variety of methods have been described to perform these measurements, but there are relatively few reports of comparative results

11:30 am Improving Platform Methods for AAV Characterization Supporting Process and Product Analysis

  • Tingting Li Senior Biopharma Application Development Scientist, Sciex

Synopsis

  • Discover analytical workflows for product and process related impurities including viral capsid purity, genome integrity and empty/full vector ratio assessment.
  • Understand how these methods can be adopted in both high throughput and quality control settings

12:00 pm Lunch & Networking

NAVIGATING COMMON INDUSTRY POTENCY ASSAY PITFALLS


1:00 pm iLite® assays for the quantification of the potency and neutralizing antibody response to recombinant AAV vectors

Synopsis

  • Highly sensitive assays, with an extended dynamic range,
    have been developed based on the iLite® reporter-gene
    technology for the quantification of the potency of AAV
    encoded transgenes regulated by photoreceptor-specific
    promoters without the need for helper virus or proteosome
    inhibitors.
  • A highly sensitive, rapid, and serotype specific assay for the
    quantification of neutralizing antibodies to recombinant
    AAV vectors based on the iLite® technology will also be
    presented

1:30 pm Lessons Learned From Potency Assays For Gene Therapy Drug Products

Synopsis

  • Quantitative potency assays were developed to demonstrate correction of b-thalassemia and sickle cell disease properties in an in-vitro cell culture system.
  • Considerations for assay development, qualification results, and redundancy to transduction efficiency methods will be discussed.

2:00 pm Development of Potency Assays for Non-Enzymatic Targets, Considerations for Method Validation & Deployment in External Testing Laboratories in the US & EU

Synopsis

  • Developing potency assays for targets that have no measurable biological activity
  • Consideration of the cell types for potency methods
  • Data analysis for GMP compliance

UNCOVERING THE PRESENCE OF CONTAMINANTS: DETECTING & CHARACTERIZING IMPURITIES

1:00 pm Characterization of AAV Genome Integrity Using 2D-ddPCR

Synopsis

  • To characterize genome integrity, a duplex ddPCR method is developed to target the 5’-end and 3’-end of the AAV genome simultaneously which measures the percentage of full genome
  • 2D-ddPCR has been demonstrated to correlate with orthogonal methods in characterizing genome integrity

1:30 pm Accelerating Gene and Cell Therapy Pipelines with Single-cell Multi-omics.

  • Yue Wang Technical Application Liaison, Mission Bio

CHARACTERIZING PARTIALLY FILLED CAPSIDS



1:00 pm Approaches to Capsid Characterization for AAV

  • Jonathan Hill Senior Scientist, Analytical Development, Solid Biosciences

Synopsis

• Characterizing your Gene Therapy Product
• Determining the correct approach
• Challenges of characterization

1:30 pm Using Characterization of Partially Filled Capsids to Determine Biological Activity

Synopsis

  • Best Practice for Characterizing Partials
  • How to determine biological activity with partials
  • Challenges of characterizing partially filled capsids

2:00 pm De-risk Development through Advanced Characterization of Capsid Proteins by Mass Spectrometry

  • Chen Li Director, Analytic Sciences , BioAnalytix

Synopsis

De-risk Development through Advanced Characterization of Capsid Proteins by Mass Spectrometry

This session will cover:

  • Advanced LC-MS Approaches for Characterization of Viral Proteins and HCPs in Gene Therapy Products
  • Product Attribute Monitoring for Viral Proteins by Intact Mass Spec and Peptide Mapping
  • Quantification and Characterization of Gene Therapy Expression Products In Vivo

2:30 pm Afternoon Refreshments & Networking

3:00 pm PANEL: Past, Present and Future of AVV Development

NAVIGATING ANALYTICAL COMPARABILITY CHALLENGES

3:30 pm Analyzing AAV Vectors Using Droplet Digital PCR

Synopsis

  • Choosing ddPCR assays for analytical development
  • Optimizing vector titer protocols
  • Characterizing vector genome integrity

4:00 pm In-process rAAV genome titer assays on QIAcuity—a fully integrated and scalable digital PCR platform

  • A’Drian Pineda Sr. Business Development Manager, digital PCR, QIAGEN
  • Hui-wen Liu Sr. Scientist, Analytical Development, Gene Therapy Franchise, Resilience

Synopsis

As gene therapy programs grow in number and scale, manufacturing processes must keep pace with demand by providing the speed, throughput, and automation necessary for advancement. Droplet digital PCR is the current gold standard for quantifying rAAV genomes but suffers from a long turn-around time and low throughput. The QIAcuity dPCR system provides a fully integrated solution that reduces hands-on time while improving throughput and turn-around time.

A comparison of in-process samples to the current gold standard system shows that:

•QIAcuity dPCR is as accurate as the current gold standard but also at a lower cost and much higher throughput

•Sample preparation and upstream optimization are key for accurate in-process rAAV genome titer quantitation

4:30 pm Chairs’ Closing Remarks