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 gene therapy and enables accurate and reliable quantitation of AAV total capsid titer. With the AAVX and AAV9 biosensors and the new high-sensitivity kit, Gator covers AAV serotypes 1 – 10 with a dynamic range from 5E+6 to 1E+13 vp/ml.
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 minimum sample volume of only 40 µl makes this kit is a valuable asset for upstream processes. Gator Bios AAV portfolio offers a single flexible technique with multiple applications in the field of gene therapy that offer high accuracy, short assay times and a wide dynamic range.
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.
Additional features
- Robust: Quantitation and Empty/Full ratio can be determined in crude extracts, cell medium or after purification.
- Low maintenance: Probes and sample plates are disposable. There is no need for extensive cleaning protocols or instrument maintenance.
- Cost effective: AAVX and AAV9 probes can be regenerated automatically to decrease the cost per assay.
- Little material needed: Quantitation can be performed with a minimal sample volume of 40 µl per sample.
- Low sample consumption: The Measurement is non-invasive. Precious sample material can be recovered after quantitation.
- User friendly: The intuitive software enables easy assay setup and analysis.
For more information, see the Application Note: Comparison of Gator® BLI and Progen ELISA for Quantitation of AAV Serotypes
Gator Bio BLI systems for AAV applications
GatorPlus
Next gen label-free BLI instrument with high sensitivity
GatorPlus is a next generation Biolayer Interferometry (BLI) system. Among the applications are label-free quantitation and kinetic analysis of antibodies, AAV, proteins, peptides, nucleic acids, aptamers, liposomes, viruses, small molecules and epitope binning/mapping. The system uses the 384-well plate format and offers best-in-class baseline stability for higher sensitivity.
GatorPrime
Next Gen label-free Biolayer interferometry (BLI) instrument
GatorPrime is a next generation Biolayer Interferometry (BLI) system. Among the applications are label-free quantitation and kinetic analysis of antibodies, AAV, proteins, peptides, nucleic acids, aptamers, liposomes, viruses, small molecules and epitope binning/mapping. This system uses a variety of new biosensors to make assay setup and performance as fast and easy and possible.
