Just a quick off-topic question which people here may have experience in:
We have a protein which expresses with a GST tag. It has much poorer expression without. When the tag is cleaved, the protein does not crash out but we are having difficulty separating it as the proteins seem to stick together. When run on a GST column, some of the protein elutes with GST and some sticks to the column with GST. We are losing over 90% in this manner. The protein expresses to ~2mg/ml but is relatively stable and has crystallized. If anyone has similar experiences that they managed to resolve(or didn't) or any suggestions, I would appreciate it.
Jon Read
Associate Principal Scientist
_____________________________________________________________________
Astrazeneca
R&D, Decs,
Astrazeneca, Mereside, Alderley Park, Cheshire, SK10 4TG UK
Tel +44 (0)1625 510661 Mobile +44 (0)7775 040376
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