Re: ABI PTH columns

Bruce A. Stanley (bstanley@psu.edu)
Thu, 18 Mar 1999 09:47:31 -0500

Has anyone out there tried "repairing" these PTH columns (which I have
never worked with)? There are lots of reasons that columns go bad, but
several of these can be remedied with a little home repair. People
generally tend to treat columns as black boxes, just replacing them when
separations go off, but opening the column, scraping off the "dirty" top
1-2 mm of packing material, and repacking new material in there often wor=
ks
wonders. Ideally this should be done with an inexpensive column packing
device (Perseptive Biosystems sells one for about $100, if memory serves
me, as do other companies, I am sure), but even tightly packing with a
spatula often works well enough to extend column life considerably, a tri=
ck
I learned years ago from Bernd Kn=F6dgen in Nikolaus Seiler's lab.

Being able to purchase the loose packing material and obtaining the corre=
ct
method for wetting it for packing is of course dependent on the particula=
r
column and the company's policies, but we have had great results working
with Perseptive and their POROS media. For our applications, side by side
comparisons of columns we packed from scratch in the lab (using our norma=
l
HPLC pumps at a little over 3000 PSI for about an hour, considerably long=
er
than manufacturer's recommendation of ~ 10 minutes) gave separation resul=
ts
equivalent to factory-packed columns, and once one gets over that initial
fear of opening a sacrosanct column, repairs are often quick and easy.
Saves both money and time waiting for a new column to be delivered, etc.,
for those of us who can't afford to have spare columns sitting around "ju=
st
in case". Note however that we have only done this with 2.0-4.6 mm bore
columns (100-250 mm length), so although it is theoretically possible to =
do
with smaller bore columns, I can't directly attest to success with smalle=
r
bore columns.

-------------------------------------------------------------------------=
------
Bruce A. Stanley, Ph.D.
Director, Scientific Programs
Section of Research Resources, Room C1734
Penn State College of Medicine H093
500 University Drive - PO Box 850
Hershey, PA 17033
Telephone and VoiceMail (717) 531-5329 --- Lab and FAX (717) 531-4055
bstanley@psu.edu
http://www.hmc.psu.edu/stanley/stanley.htm

>Well, the killer problem is with Pro-Met-Val. Once you loose that
>separation, kiss
>the column goodbye. I never saw problems with Ser-Gln-Thr. I use a
>Beckman micro-
>PTH column. Good luck. Regards. Tom
>*****************************************
>Thomas Miller
>DuPont Company
>Protein Sequencing Lab and Oligo Synthesis
>email: Thomas.J.Miller@usa.dupont.com
>phone: 302-695-1745
>
>Opinions are my own and not of my employer
>
>
>
>Dougie wrote:
>I read recently about the problems a few people were experiencing with P=
TH
>columns for the Procise. I too have been waiting for a couple of months =
for
>a new column that I ordered. I did receive a PTH column but sent it back
>just before they went onto backorder because the resolution between Q/T =
and
>K/L didn't look right. I did manage to get another column sent and it to=
o
>gives a similar profile. The Q/T separation is not great. Is it true tha=
t
>if
>a column is on its way out then the resolution between Q/T and K/L are t=
he
>first ones to go ? Again, any views or updates would be appreciated.
>yours
>Dougie lamont

-------------------------------------------------------------------------=
------
Bruce A. Stanley, Ph.D.
Director, Scientific Programs
Section of Research Resources, Room C1734
Penn State College of Medicine H093
500 University Drive
Hershey, PA 17033-2390
Telephone/VoiceMail (717) 531-5329 --- Lab and FAX (717) 531-4055
bstanley@psu.edu
http://www.hmc.psu.edu/stanley/stanley.htm