Chromatography on Heparin-sepharose

cjs at canctr.mc.duke.edu cjs at canctr.mc.duke.edu
Fri Jan 17 15:35:19 EST 1992


In article <1992Jan15.225404.4319 at usenet.ins.cwru.edu>, bl275 at cleveland.Freenet.Edu (Dan Diaz) writes:
>
>I am having a devil of a time getting my enzyme (an exonuclease)
>to behave consistently on Hep-Seph CL-6B (Pharmacia).  On some
>home-made H-S the enzyme eluted at about 250 mM NaCl.  Using the
>Pharmacia material, the enzyme elutes much later (closer to 350
>mM.  I would appreciate any email from other protein types who
>have used heparin chromatography.

I used to use Heparin Sepharose in a purification protocol and found it
to be quite variable, as well. I'm not sure what type of samples you're running
on this column, but I found that checking the conductivity of the
samples aided in the consistency of my results. However, there is always
variation between the preparations of resin, with regards to the
amount of heparin conjugated to the resin and I believe that this is
what leads  to the variability in binding and it's very difficult to
correct for this.



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