Analytics of AAV production using Biolayer Interferometry

Determination of AAV total capsid titer and empty/full ratio using Gator Bio’s biolayer interferometry (BLI) technology

Discover the flexible technique that accurately determines AAV titer and Empty/Full Ratios in purified or crude samples.

Biolayer Interferometry (BLI) is an established method that is traditionally used to study biomolecular interactions. This label-free technique measures the binding of biomolecules to the tip of a biosensor in real-time to determine kinetic rates of interaction pairs or to quantify biomolecular samples. The newest generation of AAV biosensors from Gator Bio opens up this application to the field of viral vectors and enables accurate and reliable quantitation of AAV total capsid titer. With the AAVX and AAV9 biosensors and the new high-sensitivity kits, Gator covers AAV serotypes 1 – 10 with a dynamic range from 5E+6 to 1E+13 vp/ml and a CV of <10%.

The new Empty/Full Ratio Kit is a practical and cost-effective solution for a challenging step in AAV development. With this new tool, the Empty/Full ratio of AAV samples can be determined after purification or in crude extracts with a dynamic range from 5% – 100% Full and 10% Resolution. A throughput of 8 samples in 40 minutes and a minimal sample volume of only 40 µl makes this kit not only a valuable asset in research, but also for upstream and downstream processes. Gator Bios AAV portfolio offers a single flexible technique with multiple applications in research and development of viral vectors that offer high accuracy, short assay times and a wide dynamic range.

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Download Application Note "AAV Analytics Solutions"

AAV Analytics Solutions

Great flexibility

Consistent AAV analytics during the manufacturing process is challenging due to a lack of standardized methods that can cover all process steps. Biolayer Interferometry (BLI) offers a flexible technique with various applications throughout the process including capsid titer, empty/full ratio determination and Tab and Nab assays. Gator Bio’s AAV analytics solutions offer several advantages that make them suitable for integration into manufacturing processes from start to finish: crude sample compatibility, high sensitivity, accuracy, robustness, automation, and reduced hands-on time. With Gator Bio’s AAV analytics solutions, you can expect reliable and accurate data to optimize your processes and produce high-quality products. By leveraging the flexibility and robustness of BLI, you can now streamline your AAV analytics and maintain consistency throughout the manufacturing process.

High accuracy quantitations

Gator Bio directly detects binding of biomolecules to the sensor surface in real-time, without the need for additional labels, antibodies or other indirect detection methods. Together with the new generation of precoated AAV biosensors, the high sensitivity of BLI makes the accuracy of Gator Bio superior to ELISA.

Short assay time

Measure up to 8 samples in parallel within 2 minutes. With minimal preparation time, the Gator Bio system can quantify 96 samples in under 30 minutes. The straight forward data analysis in the integrated software enables you to get results immediately after the assay.

Wide dynamic range

There is no need to dilute your samples, no matter if you are in early growth, purification or QC step. This reduces the hands-on time and eliminates errors from the sample dilution steps.

Empty/Full Ratio workflow

The approach involves three steps:

  1. AAV capture on AAVX or AAV9 probes
  2. Transfer and lysis of AAV particles to release the ssDNA
  3. Quantitation of released ssDNA with ssDNA probes

ssDNA measurement

The figure shows ssDNA binding signal for lysed AAV2 capsids at the same concentration. The ssDNA signals are proportional to the corresponding Full Ratio of the sample.

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Application note - Gator Bio AAV Quantitation