AAV MANUFACTURING METHODS

Methods for producing recombinant adeno-associated virus (rAAV) particles include: an expansion phase including increasing the number of cells in at least one culture vessel containing culture medium; introducing into the cells a first polynucleotide sequence including a transgene flanked by AAV inverted terminal repeats, and optionally a second polynucleotide sequence including AAV rep and cap genes, and/or a third polynucleotide sequence including one or more helper genes; a production phase including culturing the cells into which the one or more polynucleotide sequences were introduced and adding calcium (Ca) ions (i.e., salts of Ca2+) to the production phase culture medium; and isolating the rAAV particles.