I am new to working with viruses, and my task is to infect lung cells with pseudo-virus. The procedure is as follows, 1) Culture HEK293T cells in 35mm dish, transfect with lentiviral spike protein, 2) next day gather the medium, filter it to get rid of debris, and then add this medium into 35mm lung cell dish [the medium should contain spike protein then].
I was wondering if freezing the obtained "spike protein" medium would work. Freezing, theoretically, should not destroy the spike protein, yes? If yes, then the "infectious" medium should be ok to freeze and unfreeze.
Do you have any experience with such procedure?
Hi, I have no comment about this subject, many thanks, because you addressed me
Dear Yulia,
I used in my studies recombinant Spike protein, and my experience indicate on sensitivity of this protein on freezing (which seems to cause partially protein degradation), but I should note that it was a purificated protein using affinity chromatography.
In my opinion, it is two possible ways:
1) storage such Spike-containing medium in 4 degree (the fridge), maybe with added proteases inhibitors, but until 1-2 weeks it should be used for further analysis,
2) freezing this medium (without inhibitors, of course), but you should consider that the spike protein level in the medium can be slightly decreased or partially precipitate.
Good luck
Chris
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