for your AAV Processes
Comparability of AAV gene therapy products is becoming ever more important due to safety concerns. These need to be answered before moving to clinical trials. However, there is a lack of suitable reference material to accurately compare AAV products beetween laboratories and clinical trials.
PROGEN offers comprehensively characterized AAV empty capsids in a liquid form, which together with their high titers of more than 5.0E+12 viral particles/ml ensures the empty capsids can be used as a reference material in a variety of applications, including dot blot, western blot and ELISA.
Available for AAV1, AAV2, AAV5, AAV6, AAV8 and AAV9.
AAV Empty Capsid Benefits
Flexible use for a variety of applications: High virus titers >5.0E+12 viral particles/ml ensure the empty capsids can be used as a reference material in a variety of applications, including dot blot, western blot and ELISA.
Less preparation time: The high titer AAV empty capsids are provided in a liquid form in PBS with 0.014% Tween, 1 mM MgCl2 and 2.5 mM KCl. This means you save time preparing, there are less opportunities for errors to slip in and you can use the empty capsids flexibly across a variety of applications.
Reliably accurate: Through rigourous testing we know that our capsids have 90-99% empty AAV particles. Our empty AAV capsids were produced in the absence of a transgene. Therefore they do not carry a transgene and can be handled the same as proteins. Empty Capsids are also free from additives and stabilizing proteins.
Dependable quality: PROGEN follows a well-established quality control protocol for the characterization of the AAV empty capsid material. This ensures a comprehensive characterization and a reliable titer determination for each production. The QC protocol includes the analysis of sample purity by SDS PAGE followed by silver staining, (figure 1) analysis of the filling grade by AUC (figure 2) or an orthogonal method such as dynamic light scattering, as well as the titer determination using PROGEN´s carefully calibrated ELISA kits and internal reference material/international RSM for AAV2 and AAV8. This extensive quality control and the reliable characterization of the AAV empty capsids makes them highly suitable for you to use as a reference standard to ensure the validity of your assays.
Off-the-shelf: Ready-to-use and off-the shelf product, making sure your waiting time is as minimal as possible.
AAV Standards for ELISA
Quantification of purified AAV vector preparations is a critical step for clinical applications to minimize an immunogenic response and to maximise a gene transfer into target cells.
Here we show an example of the development of an internal AAV5 gold standard by using electron microsopy, qPCR and ddPCR. This AAV5 standard was then utilized to calibrate the AAV5 ELISA kit control for reliable and reproducible quantification of intact AAV5 capsids.
AAV Standards FAQs
The empty capsids are prepared in the absence of an ITR plasmid containing the AAV vector genome, so that no viral vector DNA can be packaged in the particles. In some rare cases the preparations may contain a very small amount of capsids in which fragments of the host cell genome and the packaging plasmids or other DNA fragments have been packaged.
The total capsid titer of our empty capsids is determined by PROGEN ELISA.
Analysis of the filling grade is performed by AUC or an orthogonal method such as UV/Vis.
Empty capsids (AAV1, 2, 5, 6, 8, 9, rh10 and rh74) are provided in liquid format in PBS + 0.014% Tween20 + 1 mM MgCl2 + 2.5 mM KCl without protein additives. The titer is greater than 5.0E+12 capsids/ml. Therefore, they can be used in several applications.
ELISA Kit Controls (AAV1, 2, 3, 5, 6, 8, 9, rh10 and rh74) are lyophilized. After reconstitution in ASSB 1x the final solution contains stabilizing protein, phenol red and ASSB 1x buffer. The titer is lot-specific around 7.9E+09-1.3E+10 capsids/ml. This product is mainly suitable for the ELISA.