Characterization of mouse Bmp5 regulatory injury element in zebrafish wound models.

Many key signaling molecules used to build tissues during embryonic development are re-activated at injury sites to stimulate tissue regeneration and repair. Bone morphogenetic proteins provide a classic example, but the mechanisms that lead to reactivation of BMPs following injury are still unknown. Previous studies have mapped a large "injury response element" (IRE) in the mouse Bmp5 gene that drives gene expression following bone fractures and other types of injury. Here we show that the large mouse IRE region is also activated in both zebrafish tail resection and mechanosensory hair cell injury models. Using the ability to test multiple constructs and image temporal and spatial dynamics following injury responses, we have narrowed the original size of the mouse IRE region by over 100 fold and identified a small 142 bp minimal enhancer that is rapidly induced in both mesenchymal and epithelial tissues after injury. These studies identify a small sequence that responds to evolutionarily conserved local signals in wounded tissues and suggest candidate pathways that contribute to BMP reactivation after injury.

Improving mAb capture productivity on batch and continuous downstream processing using nanofiber PrismA adsorbents.

Improving productivity and decreasing costs for biotherapeutic agents has been a focal driving force in the manufacturing of biologics. Advances in upstream processes have been continuously outpacing the ability for downstream operations to purify biologics, especially monoclonal antibodies. Continuous chromatography has several benefits for biologic purification including automated control, decreased labor, improved productivity, and more consistent product attributes. The goal of this study was to improve productivity and decrease costs associated with batch-mode and continuous purification processes. Productivities using cellulose nanofibers with a protein A ligand offer greater than 30-fold higher productivities than their resin-based equivalents using periodic countercurrent technology with multiple column chromatography. The smaller columns needed for convective mass transfer, faster processing times, and decreased costs allow for a more efficient mAb capture step. Additionally, high throughput purification (grams of mAbs/day) can be achieved from the scale-down model developed using periodic countercurrent technology. These advancements will help drive the evolution of downstream operations to manage the higher workloads due to increased upstream titers in a cost-effective manner.