ming at ahabs.wisc.edu
Sat Jan 21 15:37:56 EST 1995
In article <75551.plank001 at maroon.tc.umn.edu>, "David W. Plank"
<plank001 at maroon.tc.umn.edu> wrote:
> Ding Ming:
> The niche for FPLC (Fast Protein Liquid Chromatography) is providing
> a highly accurate and reproducible system for running salt gradients (e.g.
> NaCl,(NH4)2SO4) in ion exchange chromatography (DEAE, CM, MonoQ, MonoS) or
> hydrophobic interaction chromatography (phenyl sepharose). Using these
> salt buffers in your standard HPLC system will depassivate the stainless
> steel tubing and pump and eventually cause the system to breakdown from
> corrosion. In an FPLC system, the buffer is only in contact with glass
> and teflon tubing. Additionally, HPLC buffer systems such as
> Acetonitrile/TFA can be unfriendly to many enzyme activities and is
> one of the reasons that FPLC is targeted primarily toward protein
> purification (hence the name). However, it does have application in a
> variety of other areas.
Thank you for your good-wish-explanation, which I have been taught since I
was a college student in about 20 years ago. I have been working as a
protein biochemist long enough and in my previous working experience, I
had been working with Beckman's Gold, Millipore's Maximum, Shimadzu's
LC-6A, Waters' Millennium, and certainly, Pharmacia's "FPLC". If you are
going to sponsor an academic meeting on HPLC application in protein
purification, please let me know. I will be more than happy to be one of
the speakers:-) But first, let me correct your concepts in your post: (1)
Non-metallic Tubing, Column and Pump Head have been available for a long
time in various HPLC systems; (2) HPLC does not equal to Reverse-Phase
HPLC, i.e., you can certainly do ion exchange and gel filtration in HPLC.
It is nothing about your HPLC system, but COLUMNS you chose for your
system. Is this clear enough to you?
With my best respect.
University of Wisconsin-Madison
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