AAV Aggregation Analysis And Low Volume Characterization

Through innovative technology that enables particle quantitation and identification in as little as 5 µL of material, you can quickly obtain the critical AAV aggregation data you need to evaluate stability and ensure the development and manufacturing of a safe gene therapy.

Overcome a bottleneck in gene therapy development and manufacturing

We tested an AAV2 vector for aggregation under a variety of stress conditions—(a) the concentration of particles with an equivalent circular diameter (ECD) ≥ 2 μm and (b) the percentage of the membrane covered by particles.

Why Use Aura Systems for AAV Analytics?

Because you can achieve so much more insight with much less material:
  • Detect and characterize particles not measured by dynamic light scattering (DLS) or size exclusion chromatography (SEC)
  • Preserve sample, using as little as 5 μL per test
  • Quickly screen a wide range of conditions with our 96-well format
  • Obtain detailed information on particles that other methods can’t deliver, including size, morphology, count, and size distribution
  • Move quickly with an analysis time of about 1 minute per sample
  • Evaluate a wide range of particle sizes, measuring 1 μm to 5 mm with high reproducibility
  • Achieve high sensitivity because particles are imaged without the interference of buffer or matrix
  • See more detail for better particle identification with high-resolution magnification
  • Maintain compliance with the option for 21 CFR Part 11 software

Compare Stability of AAV Serotypes Under Thermal Stress

Using the Aura GT system, we observed low levels of SVPs in stock AAV2 samples. Well-defined, small to large granular particles are formed after thermal stress.

We observed increased levels of SVPs in stock AAV5 samples compared to stock AAV2 samples. Large fibrillar structures are formed after thermal stress.

We observed increased levels of SVPs in stock AAV8 samples compared to stock AAV2 and AAV5 samples. Complete sample degradation was observed as the stressed AAV8 sample denatured into a film-like matrix.

Particle Vue software can provide a quantitative analysis of the different AAV serotypes with and without thermal stress by measuring the concentration of particles > 2 µm.

Definitively Confirm Detected SVPs are Aggregated Capsid

Frequently Asked Questions

There are numerous AAV (Adeno-associated virus) serotypes that have been identified and characterized to date, with new serotypes continuously being discovered. As of the latest research, over 100 different AAV serotypes and variants have been identified, each exhibiting unique properties such as tissue tropism, transduction efficiency, and immunogenicity. These serotypes have diverse applications in gene therapy and research, offering potential vectors for delivering therapeutic genes to target tissues and organs.

AAVs (Adeno-associated viruses) are typically manufactured using recombinant DNA technology and cell culture techniques. The manufacturing process involves several key steps:

  • Vector Design: AAV vectors are engineered to contain the therapeutic gene of interest within the viral genome.
  • Cell Culture: Host cells, such as HEK293 cells or insect cells, are grown and maintained in bioreactors under controlled conditions.
  • Transfection: Plasmids containing the AAV vector genome and helper virus genes required for AAV replication are inserted into host cells.
  • AAV Production: The transfected cells create AAV particles, which are harvested from the cell culture supernatant.
  • Purification: AAV particles are separated from cellular debris and contaminants using chromatography or other purification techniques.
  • Formulation: The purified AAV particles are made into the final vector product for research or therapeutic use.

The manufacturing process is highly regulated and requires stringent quality control measures to ensure the safety, purity, and efficacy of the AAV vectors.