Protein Fusion and Purification System

NANTEL at UNCVX1.BITNET NANTEL at UNCVX1.BITNET
Thu Dec 10 08:53:00 EST 1992


From:   IN%"kristoff at net.bio.NET"  "David Kristofferson"  9-DEC-1992 19:09:59.09
To:     methods at net.bio.NET
CC:
Subj:   RE: Protein Fusion and Purification System

>I read your very interesting comments on the modified MBP system.  We are
>making extensive use of this system also and your comments are helpful to us.
>we have one major problem; after Xa cleavage, the free MBP will not
>rebind to the amylose column allowing purification of this away from
>the target protein - do you have any suggestions/experience in this area.

>Mark Bradley

The problem with MBP is that it bind very tightly to maltose and extensive
dialysis is not always enough. I read in an earlier e mail that MBP binds
to hydroxyapatite. If your protein doesn't bind to this media this might do
the trick. In my case, my bZIP protein and MBP have a very different pI and
I'm preparing to perform ion exchange chromatography.

Andre Nantel
Biology
University of North Carolina




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