Skip to main content

Cookies

This site uses cookies to provide you with a more responsive and personalized service. By using this site you agree to the Privacy Policy and Terms of Use. Please read our cookie policy for more information on the use of cookies on this website.

Top
  • COVID-19
    • COVID-19 Resources
  • Solutions
    • By Market
      • Academic Research
      • Agriculture
      • Biobanking
      • Government
      • Pharma and Biotech
      • Research Foundations
    • By Application
      • Clinical Trial Sample Management
      • Discovery, Compound Management and Biologics
      • Molecular Diagnostic Solutions
      • Regenerative Medicine and Cell Therapy
      • Sample Preparation and Lab Services
      • Sample Transport and Cold-Chain Logistics
      • Vaccine Development
    • Geography
      • Asia-Pacific
      • Europe
      • North America
  • Products
    • Azenta Life Sciences Virtual Booth
    • Automated Sample Storage Systems
      • Cold Storage (Ambient to -20°C)
      • Ultra Cold Storage (ULT -80°C)
      • Cryogenic Storage (-190°C)
    • Consumables & Instruments
      • Sample Tube Solutions
      • PCR & Microplate Solutions
      • Sample Cooling & Heating Labware
    • Informatics Platforms & LIMS/LIS
    • Integrations
    • Sterilization Services
    • Microplate Coding Services
  • Services
    • Genomics Services by GENEWIZ
    • Sample and Material Management Solutions
    • Sample and Material Storage
    • Clinical Trial Management
    • Drug Products Storage and Distribution
    • Sample Sourcing
    • Sample Prep and Lab Services
    • Lab Moving and Biological Transport
    • Business Continuity and Risk Mitigation
  • Support
    • Technical Support
    • Product Support Plans
    • Product Updates
    • Sample Tube Pack Request
    • Plate & Seal Sample Request
  • Resources
  • Blog
  • About Us
    • Events
    • About Azenta Life Sciences
    • Management Team
    • Board of Directors
    • Global Locations
    • Investor Relations
    • News
    • Careers
    • Quality
    • Certifications
  • Contact Us

Utility Navigation

  • Life Sciences Customer Login
  • GENEWIZ
  • BioInventory Login
  • Portals Login
  • Portals
  • Careers
Home

Utility Navigation

  • Life Sciences Customer Login
  • GENEWIZ
  • BioInventory Login
  • Portals Login
  • Portals
  • Careers

Main Navigation

  • COVID-19
    Vaccine Development
    Vaccine Development

    Accelerating global operations to manage and process samples critical to vaccine development.

    Azenta Life Sciences Virtual Booth
    Azenta Life Sciences Virtual Booth

    The exhibition experience to your desktop or mobile device

    • COVID-19 Resources
  • Solutions
    • By Market
      • Academic Research
      • Agriculture
      • Biobanking
      • Government
      • Pharma and Biotech
      • Research Foundations
    • By Application
      • Clinical Trial Sample Management
      • Discovery, Compound Management and Biologics
      • Molecular Diagnostic Solutions
      • Regenerative Medicine and Cell Therapy
      • Sample Preparation and Lab Services
      • Sample Transport and Cold-Chain Logistics
      • Vaccine Development
    • Geography
      • Asia-Pacific
      • Europe
      • North America
    Resources Center

    Learn about sample management best practices, gain advice from industry leaders, gain access to global regulatory and logistics guidelines and find simple support tools that can help you make better sample management decisions.

    Resources Center
  • Products
    Azenta Sample Pack and Catalog
    Sample Tube Pack Request

    Improve sample tracking, ensure sample integrity and drive process efficiency with Azenta Sample Storage Tubes. Explore the range, and select the best tube for your workflow.

    Low DNA Binding Microplates
    Low Binding Microplates

    Ultra low DNA binding microplates made from PP, fit for Next Generation Sequencing (NGS) and other sensitive applications with ultra-low DNA input. Off the shelf options. Custom solutions on request.

    • Azenta Life Sciences Virtual Booth
    • Automated Sample Storage Systems
      • Cold Storage (Ambient to -20°C)
      • Ultra Cold Storage (ULT -80°C)
      • Cryogenic Storage (-190°C)
    • Consumables & Instruments
      • Sample Tube Solutions
      • PCR & Microplate Solutions
      • Sample Cooling & Heating Labware
    • Informatics Platforms & LIMS/LIS
    • Integrations
    • Sterilization Services
    • Microplate Coding Services
  • Services
    Clinical Trial Management
    Clinical Trial Management

    Life science organizations must collect and manage large numbers of research samples to bring a new therapeutic or diagnostic to market. This is a complex process.

    Sample Preparation & Lab Analytical Services
    Genomics Services by GENEWIZ

    Comprehensive genomics services provide end to end solutions and best-in-class data quality from the trusted experts in genomics.

    • Genomics Services by GENEWIZ
    • Sample and Material Management Solutions
    • Sample and Material Storage
    • Clinical Trial Management
    • Drug Products Storage and Distribution
    • Sample Sourcing
    • Sample Prep and Lab Services
    • Lab Moving and Biological Transport
    • Business Continuity and Risk Mitigation
  • Support
    disaster recovery
    How Safe Are Your Samples?

    Plastic storage tubes are used in laboratories worldwide to store biological and chemical samples.

    Product Support Plans
    Product Support Plans

    Azenta  Life Sciences' world class service team is committed to ensuring you meet your business objectives by providing a flexible portfolio of service products designed to optimize up-time and productivity.

    • Technical Support
    • Product Support Plans
    • Product Updates
    • Sample Tube Pack Request
    • Plate & Seal Sample Request
  • Resources
  • Blog
  • About Us
    Putting a Sample Kit in an envelope
    Quality Assurance Certifications

    Azenta Life Sciences has established, documented, implemented and currently maintains a quality management system that fulfills the needs of customers.

    About Azenta Life Sciences
    About Azenta Life Sciences

    We are Azenta Life Sciences. We provide unrivaled sample exploration and management solutions to help our customers accelerate discovery, development and delivery.

    • Events
    • About Azenta Life Sciences
    • Management Team
    • Board of Directors
    • Global Locations
    • Investor Relations
    • News
    • Careers
    • Quality
    • Certifications
  • Contact Us

Blog

You are here

  • Home
  • Blog
  • Clearing Roadblocks in Early-Stage Cell & Gene Therapy Development

Clearing Roadblocks in Early-Stage Cell & Gene Therapy Development

January 26, 2022

AAV Particles

 

Under intense pressure to develop cell and gene therapies on tight timelines, biotech companies are encountering common bottlenecks for discovery and scale-up.

Therapies using adeno-associated virus (AAV) for gene delivery provide an interesting case study. AAV has many desirable properties as a therapeutic vector, but structural features of its genome can thwart traditional methods for molecular cloning and quality control.

We spoke with Dr. Conrad Leung, Fellow and Vice President of Technology Development at Azenta Life Sciences, to discuss these issues and recent innovations to address them.

 

What challenges are CGT companies facing during early development?

Many of our cell and gene therapy (CGT) customers are small biotech companies. They are in a unique spot compared to big pharma. For these small operations with a handful of employees, their biggest challenges are around how to scale things up. “How do I move it to the next level?”

Many of our customers work with us to scale up the things they need. They don’t have the staff to focus on this type of mundane work. Once they have the concept in mind—that is, they’ve designed the DNA —they outsource it to us to create the construct they need for their downstream applications. If you’re a cell or gene therapy company, you don’t want to invest in developing a vector—you want an expert to do that for you.

 

Why are some vectors for CGT particularly hard to work with?

Going back a few years, when we began providing AAV plasmids, I got a phone call from a customer saying they couldn’t sequence part of their construct called the inverted terminal repeat, or ITR. The DNA in an ITR region folds back on itself, forming a very strong hairpin structure, that inhibits a typical Sanger sequencing reaction.

To make matters worse, these regions are prone to acquiring spontaneous mutations—usually deletions—at various steps of the cloning workflow. Fully intact ITRs are important for AAV replication and packaging, so mutations can reduce the effectiveness of the therapy and increase variability in downstream results. With traditional analysis methods, scientists can’t keep close watch on these unstable regions.

Production of AAV particles for therapeutic gene delivery 

Production of AAV particles for therapeutic gene delivery. A gene of interest is cloned into an AAV vector to create a transfer plasmid, which is then introduced into a eukaryotic cell line. Generating functional viral particles requires co-transfection with several viral genes, supplied by packaging plasmids (i.e. Rep/Cap and helper plasmids). Recombinant single-stranded DNA, containing the transgene flanked by inverted terminal repeat (ITR) sequences, is packaged into capsids. The ITRs form very stable hairpins, which are problematic for amplification and sequencing of AAV plasmids during the early steps of the workflow.

 

How did you address the issues with ITR regions?

We developed a methodology to synthesize and prep AAV plasmids with higher fidelity along with a robust QC process to verify the accuracy of the construct. The game changer was a new Sanger sequencing protocol that could read the entire ITR region with a strong signal. It’s a powerful tool. You can now monitor ITR integrity with high confidence. If mutations do occur, we have an ITR correction process to fix them. These new tools alleviate a lot of headaches for researchers working with AAV, speeding up their overall workflow.

 

How do these new technologies for AAV help beyond the R&D stage?

As customers move closer to the pre-clinical and clinical phases, it becomes increasingly important to confirm the quality of their vectors. Contaminant DNA or mutated AAV sequences introduced during viral packaging can affect clinical safety and efficacy. As part of quality control, you need to measure the integrity and heterogeneity of the packaged material. As you move downstream and encounter more scrutiny from industry partners and regulatory bodies, sequencing becomes more important. Recently one of our customers reached out to us to help determine what percentage of viral purity is required for safety and efficacy. We’re currently evaluating commercial batches and working with the FDA to establish these guidelines.


Azenta Life Sciences provides a comprehensive range of solutions across every phase of CGT development. Learn how we can support the rigor of your program all the way from target discovery through scale-out.

Explore Now

 

About Conrad Leung

Conrad Leung

Conrad Leung, Ph.D., is a Fellow and Vice President of Technology Development at Azenta Life Sciences. He oversees a team of automation engineers and R&D scientists. His teams provide automation solutions to the company’s laboratories and develop innovative technology to support customers' scientific research. He has been with the company for more than 15 years.

  • Cell & Gene Therapy
  • Genomics
  • Sequencing
Home
Facebook twitter youtube linkedin
Copyright © 2023 Azenta US, Inc.

Footer menu

  • Privacy Policy
  • Cookie Policy
  • Terms and Conditions
  • Terms of Use
  • Careers
  • Subscribe