Hi,
In one of bizarre finding, I got no binding in ELISA for one set of mAb expressed transiently, however positive and negative including Test2 sample gave expected results.
Further when purified all test samples gave sharp band in SDS, it shows Biacore binding as well as ADCC activity so at chip chemistry and cell based assay if its binding then why not in ELISA.
With someones working experience and if observed same phenomenon, please help me to identify reasons?
In general unfortunately not every antibody works similarly well for every application, that is why there are often multiple ones available sometimes labeled specifically as suitable for ELISA, CHIP etc.
Most of the time we scientists just have to live with this fact and get a different antibody altogether. It is not a simple question to answer why. It can be a result of various conditions which are different. For example could the epitope simply not be exposed due to a different ion concentration, binding to the well plate etc.
Dear Thomas,
Definitely, I agree to " could the epitope simply not be exposed due to a different ion concentration, binding to the well plate etc".
Thank you for the valuable input!!!
Regards,
Santosh Kumar
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