Hi everybody,
I have transfected HEK293T cells with lentiviral producing plasmids. If I wash the cells 12 hours after transfection and replace the supernatant with fresh medium how many hours would be the best time after washing to harvest the supernatant with high virus concentration? It should be mentioned that the transduction of target cells performs 3 rounds.
I would start collecting about 24 hours after the media change (36 hours after transfection)
After transfection with Lenti plasmids we harvest the supernatant after 4 days. We measure titre of the virus using p24 ELISA and immediately transduce the target cells.
I should clarify...we start collecting at 36 hours post-transfection and do subsequent collections at 48h, 60h, and 72h.
The titre of virus was maximum 48h post-transfection. I determined the titre via transduction of HeLa cells, rather than p24 ELISA. The latter does not tell whether the viral particles are actually capable of transducing target cells.
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