How to effectively prevent protein aggregation?

Gang Dong via methods%40net.bio.net (by gang.dong from univie.ac.at)
Mon Jun 1 06:53:26 EST 2009


Dear all,

 

We are having problems to purify a protein due to aggregation. The protein
is MBP-tagged and stays in solution after the tag is removed. Limited
proteolysis suggested this protein is mostly folded, with its C-terminal 100
aa unstructured. However, both the full-length and the truncated versions
were eluted at the void volume on the S-200 GF column. We suspected that
this is due to non-specific interactions among molecules that leads to the
aggregation. Since this protein is a component of a large complex, the
non-specific interaction might be caused by the exposure of some hydrophobic
patches. Before we try out different additives, I am wondering whether any
of you have similar experience? If so, how did you solve the problem. Any
suggestions are welcome.

 

FYI, the buffer we used contains 20mM Tris (pH8), 300mM NaCl, 5% glycerol. 

 

Best,

Gang

________________

Gang Dong

Max F. Perutz Laboratories (MFPL)

Vienna, Austria 



 



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