I am trying to select hybridomas (monoclonal) specific for some antigens and I have already cloned some giving a positive response by ELISA. The problem is that, further propagation Ab production and testing of the clones is not showing a constant positive response. Do you have any suggestion? Is there more than one clone and thus I need to subclone another time?
hi
1- you make freeze from mother clones.
2- Sometimes you need subcloning at least 4 times.
Also, what happened to you is normal and would halt production or class switching there. Then you need to be careful and not to lose even a single cell clones should work to get sustainable proceed. You can also increase the efficiency of the technique by electrofusion .
good luck
How many days that hybridoma cells incubation before you take sample for ELISA test? it possible If not equal day in each test. Or you should done limiting dilution in order to select a single clone.
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