[Protein-crystallography] Re: information

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Tue Aug 15 07:14:04 EST 2006


jasmeet singh wrote:

> Hi,
> 
> I work in University of California, San Francisco.I saw your message posted
> on the web about HPLC vs FPLC. I am working with two proteins which I wanted
> to purify and so I was wondering if you could help me. My proteins are
> S100A8 and S100A9 which are 8 and 13 kda respectively. I wanted to know
> which method is better for purification in their case. Both tend to
> dimerize.

The main difference between FPLC and HPLC is column capacity, HPLC is
more for analytical, FPLC for preparative work. In addition, HPLC
columns may use finer matrices, resulting in higher resolution (number
of theoretical plates). 

Dimerisation will affect the results of size exclusion chromatography
while other techniques are less affected, i.e. ion exchange, affinity or
hydrophobic interaction chromatography. You should probably select a
buffer where the dimerisation equilibrium is shifted to either side,
that is where you have either mostly dimers or mostly monomers.



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