We have been using the ABIMED robot for two years, and were in fact the first in the US to get one. It is used routinely for low level Ag stained materials. Usually we reserve the low level stuff fr LC-ms/ms analysis using micro capillary LC on either an LCQ or TSQ. More abundant samples go to the MALDI, and often require additional cleanup by zip tip or some other technique.
It is definitely the best investment in terms of productivity that we've made. However, the interface is archaic, but has been modernized somewhat with the newer 96 well version. Another design flaw is that the original instrument had a vent fan that sucked in lab air and blew it over the samples, thus allowing dust to build up inside the instrument. What is the point of putting everything inside of a nice plexiglass case if this happens? One of the more handy people in the lab, using his basement shop, built an attachment to the back of the instrument that we connected to a HEPA air filter. Now filtered air gets pumped in and dust buildup has stopped. In the new 96 well instrument, the air fan (at least on ours) is not right either, and we will have to fix it as well. As far as the tubes goes-- one can wash them ahead of time. We run a quick 30 min wash program with the tubes on the robot, then add the gel pieces to the washed tubes. This helps to clean them, and a!
lso serves to check that the instrument and tubes are set up properly.
-Elliott Nickbarg
Genetics Inst. Inc.
>>> Len Packman <lcp2@mole.bio.cam.ac.uk> 11/18 5:10 AM >>>
Compilation of private replies:
We have a DigestPro from ABIMED but sold in the US by LC Packings. We
purchased one of the first ones commercially available. Our instrument is
set up for the digestion of 30 gel samples. I have heard that ABIMED is in
the process of coming out with a 96 sample version if they haven't already.
Here's what I like about the instrument. Using peptide mass mapping by
reflector MALDI-TOF, we have been able to detect and positively identify
proteins (BSA or myoglobin) at the 1 pmol level. That is to say, 1 pmol
loaded on a Ag stained gel. At levels below 1 pmol, it is a crap shoot.
If you just want to run the standard in-gel digest protocol supplied with
the instrument, it works very well for this purpose. The instrument itself
is very reliable. For what it costs vs. the time in labor it saves, it has
been a very good investment for us. I would also say that from the point
of view of reducing keratin contamination, the instrument design really
helps in this regard. Here's what I do not like about the instrument.
The control interface for the DigestPro is truly archaic. It uses a
controller which is in reality a 386 computer running under a DOS like
interface. This is simply not up to modern instrumentation control
standards, IMHO. The control language is cryptic and making changes to the
methods is a real pain. And, trying to write one from scratch is even
harder. The other problem we have had is the rack cover for the tubes
holding the gel slice is made out of polycarbonate or plexiglass. After a
just a few months of use, the vapors from the acetonitrile have corroded
and destroyed the plexiglass cover. This is a design flaw. Some other
material should be used for the cover. The biggest complaint I have with
the instrument is the tubes that are supplied with (coming from ABIMED) it
are contaminated with wool. We noticed that there were pieces of wool or
hair or something in a lot of the tubes. Whoever is putting these tubes
together should really get rid of that old black sweater or maybe consider
shaving the beard off. Seriously. We discovered this when our hits in the
database for our blanks came up as sheep wool keratin. We solved the
problem by using a thin stream of nitrogen gas to blow out all of the wool
out of the tubes before proceding with the digest. So, in closing, the
ABIMED instrument, despite its limitations, is about half the cost of the
competition and does the job.
***********************************************************************
We have been using the Abimed robot for a couple of years with good luck as
well, but we have just now "upgraded" to the Genomics Solutions 96 well
digestor and I am still in the process of validating it and getting the
cobwebs out of the software. We also bought their ProMS MALDI spotter
but, alas, when I went to use it for validation of the Progest, it
cratered....no response from the touch screen, no control from the attached
computer, so I rebooted...... now there is absolutely nothing on the LED
screen. So, I am hopeful they can fix it, at this point it is merely a
matter of time until I can give you a report on its performance.
We were looking seriously at the spot cutting robot as well, but it seems a
little too far away from "done" for the cash they are asking. As far as
the imaging software from them, the BioImage is rather rough around the
edges but I am not convinced that any of the currently available programs
are any better.
Well, that's my two cents, I will update you more if/when I get a little
bit more data out of these two robots.
***********************************************************************
As I know Protana (www.protana.com) has some kind of semi automated
workstation for protein digestion. It allows to run 400 samples per day by
one person. They are routinely getting results from 50-100fm BSA by MALDI
using 2-3 % of supernatant. I saw also data from 25fm analyzed by QSTAR.
They can read a complete sequence of a BSA peptide.
*********************************************************************
Dr Len C. Packman
Assistant Director of Research
Protein and Nucleic Acid Chemistry Facility
Department of Biochemistry
University of Cambridge
80 Tennis Court Road
Old Addenbrookes Site
Cambridge, CB2 1GA, UK
Tel: +44 (1223) 333639
FAX: +44 (1223) 766002
e-mail: lcp2@mole.bio.cam.ac.uk
Visit my WWW page at http://www.bio.cam.ac.uk/proj/adr/PNAC/pnac.html