An autonucleolytic suspension HEK293F host cell line for high-titer serum-free AAV5 and AAV9 production with reduced levels of DNA impurity.

ABSTRACT: We sought to engineer mammalian cells to secrete nuclease activity as a step toward removing the need to purchase commercial nucleases as process additions in bioprocessing of AAV5 and AAV9 as gene therapy vectors. Engineering HeLa cells with a serratial nuclease transgene did not bring about nuclease activity in surrounding media whereas engineering serum-free, suspension-adapted HEK293F cells with a staphylococcal nuclease transgene did result in detectable nuclease activity in surrounding media of the resultant stable transfectant cell line, "NuPro-1S." When cultivated in serum-free media, NuPro-1S cells yielded 3.06 × 10¹⁰ AAV5 viral genomes (vg)/mL via transient transfection, compared with 3.85 × 10⁹ vg/mL from the parental HEK293F cell line. AAV9 production, fol

SUBMITTER: Howe G 

PROVIDER: S-EPMC11385518 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature