Recombinant adeno-associated viral vectors are an important technology for novel in vivo gene therapeutics. This presentation will highlight a high throughput, QC-ready, platform approach to AAV characterization.

• 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

Detailed and comprehensive analysis of virus vectors are required to ensure their efficacy and safety. Recently, novel biophysical characterization methods of AAV have been emerged. Here we introduce primary structure analysis by the combination of information from CE-SDS and LC-MS, which also provides accurate ratio of VPs. Then, multiwavelength SV-AUC will be introduced that enable to identify comprehensive size distribution profile of particles with information on constituents in AAV samples.

Current manufacturing and purification processes of adeno-associated virus gene therapy products result in relatively low product yields. The battery of analytics required for complete in-process testing, product characterization, lot release, and stability testing can consume significant amounts of clinical lots. Empty capsids are a key product-related impurity for which the FDA requires batch release testing. An assay suitable for routine GMP QC testing that can accurately measure %Empty capsids in both in-process samples and DS/DP of Gene Therapy products is needed. This presentation describes a new method of quantifying empty capsids, enabling an end-to-end analytical control strategy.

Regulatory guidelines require the determination of product and process-related impurities for gene therapy products. Here, we present methods for the characterization of AAV capsids on various levels using liquid chromatography-mass spectrometry. Application of native mass spectrometry for rapid assessment of empty and full capsids will also be presented. We will also outline methods for the analysis of residual host cell proteins at various stages of AAV downstream purification.

In this presentation, we will discuss the development of a high-throughput, QC-friendly multi-attribute assessment of AAV products and process-intermediates by size exclusion chromatography.

Gathering the info you need about your AAVs chews up too much sample and time. From just microliters of AAV, Stunner gets you answers on titer, empty/full ratio and aggregation. Uncle gives a unique look at the stability of your capsids to solve capsid engineering and formulation problems. We'll look at how to gather so much info from so little sample, and how it can make your analytical strategy more robust.

Mass photometry is a novel, easy-to-use bioanalytical technology that measures the empty-full AAV capsid ratio in minutes using minimal sample amounts and without the need of sample preparation. Circumventing the requirement of large capital expense and skilled operators, it can be employed throughout the manufacturing process. We present a novel mass photometry instrument dedicated to the challenges of AAV characterization. 

Innovative analytical tools from Bio-Techne can support gene therapy workflows from discovery to quality control and address certain critical quality attributes of your therapeutic. Today’s presentation will explore the many ways viral vector analysis is streamlined with ProteinSimple instruments and how our products improve line-of-sight across the development process. 

There has been a rapid increase in the number of clinical studies of gene therapy using adeno-associated virus (AAV) vectors. There are several different AAV serotypes which have differences in their chemical and physical stability. In this presentation, we share data on the stability of AAV8 and AAV9 in different formulations under conditions relevant to manufacturing, labeling, long-term storage, and clinical use.

Human CD34+ cells are the core components of ex vivo lentiviral gene therapy (GT) for the systemic treatment of monogenic diseases. Understanding the changes occurring in hematopoietic stem and progenitor cells (HSPC) upon genetic engineering is critical in determining the potency of GT drug products. We combined state-of-the-art multiparameter flow cytometry and next-generation single-cell technologies for the comprehensive characterization of the main sources of human HSPC and of different engineering conditions.

Our cGMP production suites and single-use systems will allow the production of multiple products simultaneously at up to 3,000-liter bioreactor scale. We are developing reliable and robust production systems that can deliver higher amounts of AAV product per batch.

Implementation of a manufacturing process that assures a predefined quality of the product is a critical requirement for the licensing and marketing of every CGT product. This presentation will discuss the current challenges in gene therapy technical development to ensure safe, well-characterized products.