I have transfected BSR5-T7 cell using TransIT LT1 from Mirus Bio, and 16hrs after transfection changed the Opti-Mem used in the transfection with full growth medium containing 5% FBS. Is there a way to remove or inhibit the trypsin deactivating components of FBS from the medium as I need to use trypsin to activate the virus that is to be produced form the transfection for further infections.
Not that I can think of. Trypsin is a pretty generic protease, so to remove its activity, you need to remove proteins, which is a major component to FBS.
Hi,
Could not find an established method for this. Though you can try to find out the lowest trypsin concentration, which is not inhibited by the given media volume (or 5% FBS). Based on that you can spike your culture with desired trypsin amount.
Tryspin and the tryspin inhibitor protein (present in serum) annihilate each other in a stochiometric manner. You can simply add enough trypsin to titrate all the active inhibitor out of the serum. It will take a lot of trypsin, by cell culture standards. Make a highly concentrated stock from trypsin powder, and determine empirically how much you have to add until your virus starts getting hit. Once you have documented what amount you need, it will be easy next time.
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