Chemical additive screening in microplates
Chemical additives for screening were diluted in either sterile water or
dimethyl sulfoxide (DMSO, 100% v/v) where appropriate. Low, medium and
high concentrations for each chemical were based on available literature
describing their observed effects on recombinant protein expressionin vitro (listed in Table S1). Cells were initially grown to a
density of 4 x 106 viable cells/mL in Erlenmeyer
flasks and mid-exponential phase cells were transfected with
AAV-encoding plasmids prior to transferring to 24-shallow-well
microplates (Corning) at 2.8x106 cells per well, 20
min post-transfection. Chemical additives were added to cells 24 hours
post-transfection (HPT). Cell cultures were harvested at 72 HPT for
viral genome titre analysis by ddPCR.