Cell and Gene therapies offer tremendous promise to address human disease but their complexity and diversity present unique challenges. USP is engaging with stakeholders to identify and develop documentary and physical standards to support cell and gene therapy developers by streamlining development and increasing confidence in analytical data.  This presentation will provide an update on documentary and physical reference standards under development in areas such as plasmid DNA, AAV Empty/Full Capsid Analysis, and AAV Vector Genome Titer.

Getting almost any information on AAVs takes too much sample and time. See how the new tools from Unchained Labs help with buffer exchange and get you answers on empty/full ratio, quantification, aggregation and the stability of your capsids with teeny-tiny sample volumes.

Making changes to the manufacturing process is an inevitable part of making a better product. For gene therapy products, often the major manufacturing change involves changes of manufacturing platform and/or manufacturing site. I will discuss major challenges and technical considerations for establishing product comparability, and provide real-life examples in this talk.

• Rational approach to designing a ddPCR assay
• Improved method performance throughput method life cycle
• Analytical study to compare qPCR to ddPCR assay

Mass photometry is a rapid, labelfree, ultra-sensitive microscopy technique, which allows mass determination at the single-molecule level based on detecting scattering signal (contrast) generated by an object. We present the technology and some examples analysing biologics, including for gene therapy, measurement of AAV particles to determine full / empty ratios.

Analytical techniques based on light scattering are essential for quantifying quality attributes and other characteristics of gene therapy nanoparticles. Dynamic light scattering (DLS) and multi-angle light scattering (MALS) coupled to separation technologies (SEC-MALS, FFF-MALS) reveal molar mass and size, aggregation, physical titer, empty:full ratio and stability.  This presentation will outline light scattering solutions to industry and regulatory needs for AAVs and other DNA/RNA delivery modalities. Light scattering techniques are simple to operate and enhance productivity while directly accessing the core biophysical properties that define these CQAs.

We developed a method based on flow imaging microscopy, combined with an image classification approach, based on a convolutional neural network for the analysis of subvisible particulate impurities in cell-based medicinal products. Jurkat cells and Dynabeads were used as representation of cellular material and non-cellular particulate impurities, respectively. Our method successfully detected and quantified Dynabeads and cells with other process-related impurities, such as cell agglomerates, cell-bead adducts, and debris.

Adeno-associated viruses are the most common vectors used in gene therapy. The complexity of the AAV particles presents several challenges for the development of analytical methods aimed at the characterization of these products. Here, we explore several case studies which highlight the challenges associated with characterization of critical quality attributes, and we propose solutions to overcome them.

Cell therapy products tend to be novel and can be more complex compared to other biologics, presenting a number of analytical challenges. For example, product understanding is essential as potential therapies progress through clinical trials, however, the development of relevant bioassays to identify critical quality attributes (CQAs) and evaluate drug product potency and comparability can be limiting. Moreover, some cell-based drug products have a short shelf-life and require rapid release testing, or have small-batch sizes that necessitate alternative testing approaches. These considerations for analytical development and for the characterization of cellular therapies will be discussed.

Cell and gene therapies (CGTs) offer tremendous excitement and potential. CGTs are inherently complex and often highly variable, which represents a critical industry challenge. Developing robust, scalable processes and products are necessary to enabling clinical translation and, ultimately, commercialization. Establishing highly characterized and reliable critical raw materials will aid in reducing risk and improve overall consistency contributing to CGT commercial success and global patient access.