IUBio

Delipidation of protein solutions

Dr E. Buxbaum EB15 at le.ac.uk
Thu Sep 25 18:55:24 EST 1997


Achim Recktenwal, Ph.D. wrote:
> 
> I am looking for methods to delipidate protein solutions, or protein &
> cell-suspensions after cell-disruption. These methods should be scalable
> for use in a large-scale protein purification process.
> 
> I am having big problems with the fowling of my first
> chromatography-column by lipids in the applied cell-homogenate.

A method I have used for the purification of Hsc70 is fractionated
ammonium sulfate precipitation. Lipid and other particulate matter is
removed with the first cut (at about 25% saturation) by a low speed spin
(10 000g, 10 min). This was much easier (and more complete too) than 
doing a high speed spin (100 000 g, 45 min) without AS. Of course, your
protein has to be soluble and stable under these conditions and you can
not use ion exchange for the first column.



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