I'm cloning codon-optimized scFv/FAbs into E. coli expression vectors (pBAD and pET). A coworker has been warning me of the genetic instability of plasmids containing antibody genes warning "be sure to plate everything at room temp or else disaster may occur!" I found one older reference, a book chapter from the 90s, that warns of instability of Ab genes and advises incubating plates and overnight cultures at 25C. More recent references are using 37C for plates/overnights and expressing proteins at a variety of temperatures (18-37C). Are there issues of plasmid instability associated with antibody genes?
It may have to do with getting correct folding of the antibody. I have also heard if people expressing antibodies in yeast since it is a eukaryotic cell, rather than using bacterial expression. Usually, most people try a variety of methods to get optimal expression and correct folding of their protein.
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