Hi,
Yes, it can be done by yourself. I repacked our 26/60 column a few years ago
with homemade apparatus. The column has been working for us since then.
Basically the loading apparatus was a tubing connecting the top of the
column and bottom of a reservoir (a flask with an opening near the bottom)
that contained 30%-50% suspended beads. The length of the tubing was about
2m, and I left the reservoir on the highest shelf I could find in our lab,
so that the gravity of the buffer in the column and tubing will drive the
flow in a constant even manner. It took overnight to get the column nearly
filled.
The gel filtration columns need to be tightly packed with no discontinuity.
As I remember, the GE gel filtration packing manual said: push the top
filter 2mm further down, after the column has been fully compressed by
flowing buffer at 1cm/min flow rate.
The beads need to be gently suspended in the beginning, but constant
stirring is not necessary. The flow of the buffer will carry beads into to
column (because my reservoir has an opening at the bottom). The difficult
part is at the end, when the column is filled to the top. The beads will be
compressed quite a lot when you start pumping buffer, then you need to fill
in more beads. As I remember, I removed the upper filter on the inlet
connector so that I could load beads through the upper inlet tubing with a
10mL or 30mL syringe, which generated enough pressure while loading the
beads - so that I didn't have to take the top off again and again.
There is a manual from GE on how to pack superdex columns. You should be
able to find it online. The Superdex beads used in different sized columns
from GE have different diameters. The bigger the column is, the larger the
beads are. And the larger the beads are, the easier the packing will be.
16/60 should still be fairly easy to pack. I checked the theoretical plate
number of our 26/60 after repacking, it was >11000, not as high as the 13000
stated in the 26/60 new column manual, but it was good enough for us.
Zhijie
--------------------------------------------------
From: "Peter Hsu" <hsuu...@u.washington.edu>
Sent: Thursday, July 12, 2012 9:51 PM
To: <CCP4BB@JISCMAIL.AC.UK>
Subject: [ccp4bb] offtopic: packing gel filtration columns
Hi all,
Sorry for the slew of offtopic posts, but does anyone here have any
experience repacking the large 120mL Superdex75/200 columns? Any
advice/tips on doing it? I've got an older column that's gotten clogged
while washing w/NaOH (can't go over 0.1mL/min w/o getting overpressure
alarm), and not sure the bossman would be thrilled w/buying another
column.
Thanks for any ideas.
Peter
