Applying High Throughput Processes To Optimize AAV Titers

By Eva Fong, Principal Scientist, MilliporeSigma

Of the two techniques used for cell culture, some therapeutic innovators may opt for an adherent platform, reasoning that this proven method of virus production, while less efficient to scale, is easier to move through development as compared to suspension. That said, a growing number of biopharmas have recognized the value in moving to suspension as early in development as possible in order to optimize production and establish a process that is more scalable, consistent, and cost effective in the long run.

Recombinant adeno-associated virus (AAV) has emerged as a leading modality for in vivo gene therapy. Presently, there are several approved AAV-based therapeutics on the market and an increasing number of AAV clinical trials underway. Notably, our development teams at MilliporeSigma have established an innovative approach to upstream optimization for these therapeutics, demonstrated via high-throughput methods for transfection optimization and scale-up of an optimized process to bench-scale bioreactors. Our identified process can be applied to multiple AAV serotypes and exhibits genome titer increases of between 50 and 60 percent. We recently applied this optimized approach to a customer project using their plasmids for production of AAV9, and were able to scale up the process to 3L bioreactors, and obtain a genome titer exceeding 8 x 1010 gc/mL.