If the fusion protein forms inclusion bodies, can I try to purify protein under denaturing conditions?

Purification and cleavage of the C-terminal fusion (MBP-Sce VMA intein-CBD fusion) protein has been tested under various urea concentrations. 1-4 M urea allowed 80% binding efficiency while 5-8 M urea showed 30-50% binding efficiency. The efficiency of cleavage with 30 mM DTT in 2 M urea is at least 50%, in comparison to reactions under native conditions.

Based on these data, chitin chromatography and on-column cleavage may be carried out in 1-2 M urea. Since DTT may cause rapid cleavage of the fusion protein under denaturing conditions, following the wash step the column flow should be stopped and the DTT stock solution should be added directly to the column. More.

I cant really gove you an answer,but what I can give you is a way to a solution, that is you have to find the anglde that you relate to or peaks your interest. A good paper is one that people get drawn into because it reaches them ln some way.As for me WW11 to me, I think of the holocaust and the effect it had on the survivors, their families and those who stood by and did nothing until it was too late.