I have a peculiar problem with regards to a monoclonal Ab. This Ab recognizes
specifically my target protein (and no others) only when that protein is
present in sufficient amounts ie detectable by coomassie blue staining. I have
recently purified my monoclonal Ab from a spinner culture supernatant by
passing the supernatant over an affinity column which has Ab bound to it that
recognizes the kappa light chain of mouse monoclonals.
When I ran the eluted products on a PAG only light chains could be visualised
on a coomassie blue stained gel. I know that the column is not stripping the
light chains from the heavy chains becuase I passed ascites fluid of a
different MAb over the same column and the heavy chains were easily detectable.
This would mean therefore that I have seen specific binding of light chains
only, to a target protein.
I am interested in all feedback that I may get with regards to this problem. I
am not an immunologist, but one very frustrated Ph.D student.
botdjl at lure.latrobe.edu.au