The best way to "recover" the hybridoma is through limiting dilution
cloning. 90% of the daugher clones should be positive for a stable cell
line.
You can do this in 96 well plates, soft agar or methylcellulose - whichever
you are most confortable with. Select the top two/three producing cell
lines from the cloning, expand and freeze.
As for the reason, there could be several. The clone was not monoclonal to
begin with. Some cell lines are less stable than others and will require
repeated cloning. The clone was cultured under stressful conditions -
allowing to get overly dense, grown past stationary phase to low viability,
kept in culture many, many months. These are just some examples. If this
is a clone that you need lots of antibody from, your best option is to
freeze large numbers of vials (after stabilizing), thaw when you need it,
then discard
-ned
"renjiaqiang" <renjiaqiang at sina.com> wrote in message
news:20011124175114.5148.qmail at ww02.jatek.com...
> Hi:
> I am generating a mAb using hybridoma technology,however ,the cell
> secreting antibody becomes unstable and the titeration was lowered,can
> anyone tell me the reason and provide a solution to me. I am very grateful
> to him/her.
>>> <http://www.biowww.net/forum/read.php?f=3&i=702&t=702>
>
