ANHYDROBIOSIS - FROM PROTEINACEOUS PHASE TRANSITIONS TO FREEZE-DRYING OF RED BLOOD CELLS
Dr. Michael A. Menze - Professor and Associate Dean Research and Innovation, University of Louisville
The emerging role of proteinaceous liquid-liquid phase separation (LLPS) in the water-stress tolerance of animal cells will be discussed based on recent findings on two late embryogenesis abundant (LEA) proteins that are abundantly expressed in the anhydrobiotic stage of the brine shrimp Artemia franciscana. Both these proteins form proteinaceous protein droplets in vitro and biomolecular condensates (membraneless organelles) if ectopically expressed in cells. The propensity to selectively incorporate target proteins expands the range of mechanisms by which LEA proteins are thought to confer water-stress tolerance. Furthermore, recent progress in the dry preservation of red blood cells will be presented.
STABILIZATION OF ADENO-ASSOCIATED VIRUS (AAV) VECTOR FORMATIONS BY LYOPHILIZATION
Dr. Tim Menzen - Chief Technology Officer, Coriolis Pharma Research GmbH
Adeno-associated virus (AAV) is a promising vector for gene therapy. Liquid AAV formulations typically require storage below -60 °C to provide long-term stability. In contrast, freeze-dried formulations could enable storage at 2−8 °C. Different conditions for freeze-drying of AAV8 were evaluated and undesirable instability can be significantly reduced if secondary drying is performed at lower temperatures, kept as short as possible, and the residual moisture is kept between 1.5-2%. Moreover, a promising combination of phosphate buffer, trehalose, hydroxyectoine and poloxamer was found. These results pave the way for future optimizations of freeze-drying processes for AAV vector-based gene therapy products.