Aurintricarboxylic acid increases yield of HSV-1 vectors

Pechan, Peter; Ardinger, Jeffery; Ketavarapu, Jyothi; Rubin, Hillard; Wadsworth, Samuel C; Scaria, Abraham

Effect of aurintricarboxylic acid (ATA) residues in herpes simplex virus-1 (HSV-1) stocks on the production or recombinant adenoassociated virus (rAAV) virions. The rAAV-GFP vector was produced by coinfection of rHSV-rep2/cap2 and rHSV-EGFP vectors in human embryonic kidney (HEK)-293 cells 60-mm plates. ATA was shown to slightly increase rAAV yields (DNase resistant particles (DRP) per cell) when 10 µmol/l ATA was spiked directly into HEK-293 cell media during 2 hours of HSV coinfection step. This effect was not observed when the ATA concentrations in HEK-293 cells during 2 hours of HSV coinfection were higher than 15 µmol/l. Results are representative of two independent experiments (n = 2) and are expressed as mean + SD of rAAV (DRP/cell. eGFP, enhanced GFP, GFP, green fluorescent protein.