The recombinant adeno-associated virus (rAAV) vectors used in gene therapy are usually produced by transfecting three different plasmids (Adenoviral helper plasmid (pHelper), AAV rep/cap plasmids (pRepCap), and Transgene plasmid (pAAV-GOI)) into human embryonic kidney 293 (HEK293) cells. However, the high proportion of unwanted empty capsids generated during rAAV production is problematic. To simultaneously enhance the genome titer and full capsid ratio, the ratio of the three plasmids transfected into HEK293 cells was optimized using design-of-experiment (DoE).
View Article and Find Full Text PDFWith the advent of novel therapeutic proteins with complex structures, cellular bottlenecks in secretory pathways have hampered the high-yield production of difficult-to-express (DTE) proteins in CHO cells. To mitigate their limited secretory capacity, recombinant CHO (rCHO) cells were engineered to express Blimp1, a master regulator orchestrating B cell differentiation into professional secretory plasma cells, using the streamlined CRISPR/Cas9-based recombinase-mediated cassette exchange landing pad platform. The expression of Blimp1α or Blimp1β in rCHO cells producing DTE recombinant human bone morphogenetic protein-4 (rhBMP-4) increased specific rhBMP-4 productivity (q).
View Article and Find Full Text PDFEur J Cancer Care (Engl)
September 2019
Objective: This study aimed to determine the feasibility and benefits of a combined programme of exercise and play for childhood cancer survivors on health-related quality of life (HRQOL), post-traumatic growth and physical strength levels.
Methods: Six childhood cancer survivors participated in the 8-week intervention consisting of supervised play and exercise sessions two times per week. The participants performed joint exercises, independently, at home, on the 5 days that they were unable to participate in group exercises.