We have transfected MDCK cells with plasmid DNA constructs expressing recombinant protein. We have analyzed the populations by immunofluorescence microscopy and have some variation in expression. For final purification of the recombinant protein, we would like the cells to be growing in serum free conditions. Would you recommend to isolate a high-expressing clone before OR after adapting the cells to serum-free conditions.
I would suggest to adapt the cells fist and select cells that express the recombinant protein at the highest second. The cells may lose or change their ability to produce the recombinant protein after the adaptation. Good luck
Does the plasmid contain any selection in mammalian cells? Would u mention your method for making a stable cell. I prefer to first select the clone if ur plasmid contains selectable marker. then go for adapting. For adapting the cell u may go down gradually with the serum concentration and completely off the supply.
In principle both ways should work. But you have to make sure whether you want single cell clones (clean but artificial), or a population (heterogenous but more "natural").
If you go for populations it's ok to select first and then adapt. If you need singe cell clones I would rather adapt first and select afterwards.
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