7:50 am Chair’s Opening Remarks

EXPLORING ADVANCED ANALYTICS IN THE PURSUIT FOR QUANTITATIVE ANALYTICAL RESULTS WITH LOW VARIABILITY

8:00 am In-Process Analytics of AAV vector capsid production

  • Ivana Petrović Koshmak, PhD Head of Upstream Process Development, Department for Process Development, BIA Separations, a Sartorius companyI)

Synopsis

Abstract of our presentation: In-process analytics of vector capsid production is a critical optimization target in development of AAV-based gene therapy products. In this presentation we introduce a fast at-line HPLC based system that enables differentiation of vector capsid and empty capsid production in transfection mixtures. Case study results will be shown demonstrating the effects of different process variables on capsid production and assembly. Insights about purification will also be discussed.

8:30 am Exploring the Evolution of Analytical Techniques for Gene Therapy Drug Development

Synopsis

  • Welcome to the digital age!?
    • Switching from PCR to ddPCR
    • Changing from gel to capillary
  • Old but still Gold!? – Analytical Ultracentrifugation

9:00 am Introducing Lonza Library of pre-developed, fast-qualify assays to expedite and harmonize development of cell & gene therapies

  • Behnam Ahmadian Global Head of Process Development, Cell & Gene Technologies , Lonza

9:30 am Joint Q&A Session

10:00 am Morning Refreshments & Speed Networking

Synopsis

This session is the ideal opportunity to get face-to-face time with many of the
brightest minds working to advance gene therapies. Benchmark against the
industry leaders and establish meaningful business relationships to pursue for the
rest of the conference and beyond.

BIOASSAYS & MOLECULAR BIOLOGY

OPTIMISING ASSAY DEVELOPMENT,
QUALIFICATION & VALIDATION

11:00 am Immunoassays for AAV Capsid Titre Quantification

Synopsis

  • Comparing ELISA with alternative immunoassay methods
  • Building analytical target profile for capsid titre quantification
  • Testing in-process samples and ensuring serotype-specificity

11:30 am AV Antibodies for Capsid Variants

Synopsis

  • Bottleneck: AAV antibody affinity
  • AAV capsid variants – considerations for ELISA development
  • Reliable standards for AAV variants

12:00 pm Critical Aspects of Gene Therapy Method Validation

Synopsis

  • Developing a well-controlled method that works at every site
  • Developing the reagents that are needed for success
  • Deciding when to validate and then when to revalidate
  • Managing transfers, validations and life cycle at remote sites

12:30 pm Joint Q&A Session

ANALYTICAL CHEMISTRY

DETECTION & SEPARATION OF PARTIALLY FILLED CAPSIDS

11:00 am What is Hiding in Partially Filled AAV Particles?

  • David Dobnik Senior Research Associate, National Institute of Biology

Synopsis

  • Different fractions of AAV vectors after ultracentrifugation were characterised
  •  Particle number, vector genomes and presence of impurities was evaluated and compared between fractions
  • Long-read sequencing approach can provide additional information on particle content

11:30 am Delving Deeper Into AAV Attributes: Enhanced Characterization Using multiple detection SEC and DLS

  • John Stenson Product Manager Nanomaterials, Malvern Panalytical

Synopsis

  • Enhanced characterization of rAAV vectors for viral titre and stability critical for quality of final product
  • Tracking particle size, aggregation state and capsid or particle count in downstream processing AAV samples with multi-angle DLS (MADLS) and multiple-detection SEC
  • Informing AAV potency by measurement of % full rAAV particles with multiple-detection SEC
  • Generation of multi-parametral stability profile of rAAV with DLS and multiple-detection SEC.

12:00 pm How High-throughput Sequencing can be used as tool for AAV vectors characterization and optimization

12:30 pm Joint Q&A Session

1:00 pm Lunch & Networking

OVERCOMING POTENCY ASSAY CHALLENGES

2:00 pm Potency Assays: Few Lessons Learned

Synopsis

  • Unexpected Potency Trends
  • System Suitability Criteria, how not to design them
  • Key reagents management

2:30 pm Advances in viral vector titer and impurity analysis with microfluidic, nanoliter-scale immunoassays

  • Linda Klauss Field Application Specialist, Gyros Protein Technologies

Synopsis

  • Immunoassays are commonly used for viral vector titer and impurity analysis during bioprocess development, but drawbacks in plate-based methods prevent their full implementation.
  • Immunoassay data for vector titer (AAV multi-serotype and lentiviral) and impurity (HEK 293 HCP and benzonase) analysis, using Gyrolab® microfluidic, CD-based immunoassay platform demonstrate significant improvements in analysis speed, dynamic range, and sample volume consumption.
  • These data support the implementation of Gyrolab technology in quality control and during bioprocess development of cell and gene therapies.

3:00 pm Joint Q&A Session

THE AGE-OLD CHALLENGE: EMPTY-FULL CHARACTERISATION

2:00 pm Assessing Best Practice Analytical Tools for Characterisation of Empty/Full Capsids

Synopsis

  • Methods for Empty/Full AAV capsid characterisation
  • Pros and Cons of each method
  • Orthogonal approaches

2:30 pm Treat yourself to AAV empty/full and titer data without the hassle

Synopsis

• Get answers on empty/full ratio, aggregation and titer
quantification using just 2 μL of sample, in less than a minute
– way faster than AUC, ELISA and ddPCR.
• Learn how Stunner’s unique combination of UV/Vis, DLS and
SLS can characterize your AAVs

3:00 pm Joint Q&A Session

3:30 pm Afternoon Refreshments & Networking

BEST PRACTICES FOR DEMONSTRATING COMPARABILITY

4:00 pm Solutions for Gene Vector Characterization

4:30 pm Comparability Assessment At Various Stages Of Product Development

  • Jaap Twisk Senior Director Analytical Development , UniQure

5:00 pm CMC and Analytical Development To Meet Regulatory Expectations On Gene Therapy Comparability Studies

Synopsis

  • Navigating the regulatory pathway on gene therapy comparability
  • Challenges of comparability studies on gene therapy products
  • Evaluation and analysis of Critical Quality Attributes for Gene Therapy products

5:30 pm Characterization of RNA-loaded Lipid Nanoparticles by AUC

  • Amy Henrickson Doctoral Candidate, Department of Chemistry and Biochemistry, University of Lethbridge, on behalf of Beckman Coulter

Synopsis

Lipid nanoparticles (LNPs) are increasingly used to deliver drugs, however, current characterization methods such as cryo-TEM and dynamic light scattering lack the necessary resolution to differentiate between loaded and empty formulations. Here we demonstrate an analytical ultracentrifugation method, using multi-wavelength detection and heavy water density matching to characterize LNP formulations. With this method we not only determine the sedimentation and diffusion coefficients, but also optically separate and identify the signal contributions from the RNA and lipid components. The density matching approach allows us to determine partial specific volume distributions, which provides information about the density of the LNPs. The density profile of LNP formulations is directly proportional to the drug loading of LNPs, and provides a high-resolution description of LNP drug loading. The same approaches are suitable for measuring the DNA loading of viral vectors and promise to provide much greater precision and accuracy to quantify drug loading of gene therapy drugs.

6:00 pm Joint Q&A Session

  • Francesco Lanucara Analytical Development & CMC, Allergan
  • Jaap Twisk Senior Director Analytical Development , UniQure
  • Kevin Jackson Technical Director, Wyatt Technology
  • Amy Henrickson Doctoral Candidate, Department of Chemistry and Biochemistry, University of Lethbridge, on behalf of Beckman Coulter

6:20 pm Chairs’ Closing Remarks