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FPLC (Repost)

Cormac Shaw cshaw at ollamh.ucd.ie
Tue Mar 17 07:35:16 EST 1998

[Apologies if you already got this but the gremlins were in the 
college mail system last weekend and it never appeared on my bionet 
list -C.]

On 12 Mar 98 at 6:30, Caroline J. Walker <walkerc at CLEMSON.EDU> wrote:
>We are buying an FPLC and are comparing the Biorad Biologic system with the
>Pharmacia FPLC system.  There are some small differences on the specs and
>the prices are about the same so we could use either. What I would like to
>hear is if anyone has feedback on how easy either of these systems is to
>use.  Is the Pharmacia software (OS/2) OK, or is the Windows 95 with the
>Biorad better?  How are the leaks? Design faults?? Reliability? Service?
>	Would be happy to hear any comments.

For your consideration:
Our lab took delivery of a Biorad Biologic system about 18 months ago 
to compliment a 10 year-ish old Pharmacia FPLC system. Now the older 
system was a manual one with a basic gradient controller and a 
hardcopy-only UV output so the comparison is a little uneven.

Both systems have their good and bad points. The more recently 
designed Biologic has more user friendly tubing connections, column 
grips, etc. which makes setup and rearrangments easier, especially 
putting connectors on tubing. Its also very easy to use non-Biorad 
columns with the supplied connection adaptors. Changing buffers is 
also much quicker than the Pharmacia system.

But not all is perfect. As another poster mentioned, air bubbles are 
a pain. Unlike the Pharmacia system, the buffer uptakes are light and 
springy and must be carefully positioned to avoid popping out of the 
liquid. The long and spongy nature of the intake covers mean that 
they draw air down to the inlet even when there's several cm of 
buffer covering them (this obvious design blip may have been 
corrected by now). The upshot is that air enters the pumps. In the 
Pharmacia system, the pump chambers are made of glass which makes 
spotting major air problems easy - a quick buffer change will usually 
cure the problem. The enclosed nature of the Biologic pumps means 
that you only spot air bubbles when the flow to your column is 
irregular and your fractions are all different volumes!! Again, a 
buffer change should do the trick but you won't be sure until you run 
the column again!! TIP: Air bubbles can also appear in the UN monitor 
chamber after buffer exits the column even when you're sure you've 
degassed well. However, this is easily solved by installing the 
supplied little gizmo for maintaining back pressure at this point.
The air bubble susceptability is a drawback of the Biologic but it is 
avoidable once you become familiar with the system's foibles. It 
could be a problem though if the machine is going to be used by a 
large number of people who won't always be familar with its pitfalls.

I can't give a comparison of the two software packages because I have 
only seen that of the Biologic but I'll make some comments on that.
The Biologic software is based on Microsofts Access database app and 
the first impression is of a nice neat layout which new users should 
find easy to learn and intuitive to use. But (there's always a but) 
again its not perfect. Inputing and saving a gradient program is easy 
and quick but, once done, if you make any changes, you must save them 
as a new protocol which soon leads to an excessive no. in the 
database (Minor complaint). More fustratingly though, oncw you've 
clicked 'Run' and a gradient operation is in progress, you can't do 
anything else with the application (e.g., copy the data from a 
previous run to a floppy for inclusion with that report that's due 
first thing tomorrow!) and you can't run another application (e.g. 
Excel) for processing you data while the Biolgic app is running. 
This, I think, is pretty crappy.

Again, the software may have been improved since we got our 
package (which was a (rather slow) 486 processor running Win3.1).

As I said, I have no idea what the Pharmacia software is like to use. 
For a dedicated desktop machine attatched to lab equipment, I 
wouldn't worry about the OS however. Even if IBM decided to throw 
OS2 in the bin tomorrow the software installed on your machine will 
still work and will still do the job as well or as badly as it would if 
OS2 had 95% of the market. Well established fact: the quality of a 
computer OS is completely unrelated to its popularity! If they want 
to stay in business, Pharmacia will still support their software 
and all those other OS2 users are out there too (you are out there, 
aren't you?). I presume, though I don't actually know, that 
transferring data from the FPLC software to, say, a spreadsheet on a 
Mac or a Wintel has been thought of and is pretty straightforward but 
maybe I'm being naive!

Some more thoughts:
The Biologic has a more logical layout and a smaller 'footprint' which 
is important if room is limited.
Biorad supplies/parts are generally cheaper than Pharmacia's but that 
may be a different story in your part of the world.
Reliability/Servicing: The FPLC is still functional after 10+ years 
of postgrad abuse though it has had some surgery. The Pharmacia 
service guy was, I think, surprised that it was still going. The Biologic 
has only been here 18 months and has not been in constant use but 
as yet, has been well behaved.

I'm tired now. I think I'll stop.

Cormac Shaw, BSc,                 |
Dept. of Industrial Microbiology, |  mailto:Cormac.Shaw at ucd.ie
University College Dublin,        |   phone: +353 (1) 706 1307
Dublin 4, Ireland                 |     fax: +353 (1) 706 1183
"Under the most rigorously controlled conditions of pressure,
temperature, volume, humidity, nutrients and other variables,
the organism will do as it pleases."

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