Next Generation Cell-line Development and Recombinant Adenoassociated Virus Production
From an informative selection of insights across upstream and downstream bioprocessing and vector manufacturing, we review the most watched presentation in the In Vivo Gene Therapy track.
Sam Wadsworth (Ultragenx Gene Therapy) began by highlighting the features of his company’s gene therapy manufacturing processes based on human embryonic kidney (HEK)293 suspension/plasmid transfection and HeLa producer cell line (PCL) suspension/adenovirus helper platforms.
In a collaboration with Bayer, the company has conducted engineering improvements to its HeLa PCL platform.
Specifically, the company made molecular changes to the plasmid components and multiple process and screening changes to the original “HeLa 1.0” platform.
The improved “HeLa 2.0” stage has a 10× increase in yield and a streamlined clonal selection.
The technology now is used for the company’s work in finding a gene therapy for Wilson’s disease.
Wadsworth showed how further improvements to the platform (“HeLa 3.0”) could be used to increase production of adenoassociated virus (AAV) vector yield and how the use of a siRNA knockdown of specific genes could improve AAV production titer.
He highlighted work on Wilson’s disease potency assays and the use of clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated protein 9 (Cas 9) to delete ATP7B (the loss of this function causes copper toxicity in Wilson’s disease) in the HepG2 cell line.
