FPLC Columns
gsarath@unlnotes.unl.edu
Tue, 29 Jun 1999 15:56:45 -0500
Dear Aki: If you packed the FPLC column with 5u particles using pumps, then you
should not experience any outgassing. Bubbles are a problem principally in open
column chromatography, where there is a large bed volume and lots of buffer.
For most medium and high -pressure packing available these days, I have yet to
encounter problems with temperature, regardless of how the columns were packed.
More importantly, these perturbations will affect size-exclusion chromatogarphy
much more than ion-exchange or other surface reactive processes. Unless you
have a very large column or some large pored matrix, you should not experience
difficulties. Bubbles in the column will cause difficulties such as poor
resolution, changes in retention due to channeling and low binding and sometimes
voiding at the top and occasionally collapse of the column matrix. I will be
happy to learn about your success with the column you packed. Cheers, Gautam