I have a similar problem Matthew.
Just got some NVoy from Novexin in which some claim can help if
hydrophobic patches are the main problem. Will see.
Regards,
Roberto
On 24 Apr 2008, at 12:35, Bottomley, Matthew wrote:
Dear All,
I have a 50kDa protein that is soluble and monodisperse at up to
approx 1mg/ml (after Ni-affinity and size-exclusion chromatography).
However, it aggregates (probably both via disulphides and via
'sticky/hydrophobic patches') when I concentrate it towards 2-3mg/
ml, even in the presence of several detergents. I don't want to add
DTT since my protein should have several intramolecular
disulphides....although I do have 2 free Cysteines, partially
exposed. I have already tried mutating the Cysteines, with little
improvement.
Any suggestions for obtaining 5-10mg/ml?
Does anybody have good experiences with usin L-Arg and L-Glu (e.g.
At 50mM) to aid concentrating (as in the Golovanov AP paper, JACS,
2004, pages 8933...)
Thanks for any input!
Yours,
Matt
====================================
Matthew J. Bottomley, Ph.D.
Senior Research Biochemist
IRBM / MRL-Rome
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