The Characterization of Adenovirus 5 Reference Material (ATCC^® VR-1516™)

First-generation adenoviral vectors represent some of the most utilized recombinant vectors that are employed in a diverse range of biomedicine fields, such as cancer therapy, gene therapy, and vaccine development. Many of these vectors are based on adenovirus type 5, and are useful for their high transgene packaging capacity, recombination ability and capacity to grow to a high titer. Adenovirus Type 5 Reference Material (ATCC® VR-1516™) is derived from a wild-type adenovirus (ATCC® VR-5™) and was first generated as a product of the Adenovirus Reference Material Working Group (ARMWG). The development of this reference material was recommended in the January 2002 report of the NIH Recombinant DNA Advisory Committee. It is to be used to establish a common reference standard to define the particle counts and infectious units for adenovirus-based gene vectors.

Replenishment batches of VR-1516™ have been developed using state-of-the-art scale-up technologies, concentration, and purification processes and optimized cryopreservation and vialing. This material has then undergone rigorous characterization of the virus particles, infectious titers and kinetics, genome concentration and authentication, residual host cell protein and DNA concentrations and substrate endotoxin and BSA titers. Final release criteria for the reference material also include tests for sterility and mycoplasma. Using novel technologies that were not available with the first production batch of VR-1516™, we have enhanced the characterization of the replenishment material. These technologies include the use of Laser Force Cytology to accurately assess the infectious titer of the adenovirus material, and Real Time Cell Analysis assays to determine viral infection kinetics.

As an important standard for adenovirus vector-based development, a well-characterized reference material is a necessary requirement to ensure accurate and safe adenovirus vectors in cell and gene therapy applications. This poster will outline the new method validation and results for new batches of VR-1516™.