antibody affinity chromatography question

Hi there,
anyone here in biology has experience with affinity chromatography?
I'm
having problem with my columns recently. my columns worked all right
before, but suddenly didn't enrich proteins. I confirmed the starting
and eluting conditions, and it should be always the same. I heard
before that these antibody columns have a definite life, such as a
number of cycles it can run, but it's unlikely two of my columns stop
working at the same time---one column was very new, and only ran a few
times.

anybody can give a clue? I'm not a prof bio and this
topic is hard to google out. hope some expert friends can share me your
valuable opinions and suggestions (why this happens... solutions...).
much appreciated!

In article <[Only registered users see links. ].net >, [Only registered users see links. ] wrote:

Not enough information. Type of antibody linkage, how the columns
were used, regenarated and stored before, and exactly what it is
that you are trying now - all this is essential to even start guessing
what might be going on.