hi, as say Zdeno protein extraction is by addition of chaotropes. but in chromatography steps chaotropes must be eliminated.

Hi there,

Solubilisation by chaotropic agent is the first step but then I guess you will intend to renaturate your protein. Renaturation is the critical step for which you need to remove the chaotropic agent and favor protein refolding in a suitable buffer, differents strategies can be considered : dialysis, dilution or on-column refolding... Hope it will help...

Hi Mohammad, as the above answers demonstrate, you can get soluble proteins from your inclusion bodies by using harsh chaotropic agents, but don't expect your protein to be functional after this treatment.

To get functional proteins, you can try to harvest them before they go into inclusion bodies.

After induction, you can take samples at various timepoints and see where you have maximal functional protein expression before the protein goes into inclusion bodies.

I agree with Steingrimur Stefansson that it would be worthwhile to invest some time investigating conditions that yield soluble protein during expression. Try different expression strains, lowering the inducer concentration, and lowering the temperature in which the cells are grown during induction (I'm supposing you are using a bacterial expression system). If you know of a chemical compound that binds to your protein and that can get into the cells, try adding it during expression to stabilize the protein as in the following paper:

https://www.readbyqxmd.com/read/25240855/overexpression-of-pseudomonas-aeruginosa-lpxc-with-its-inhibitors-in-an-acrb-deficient-escherichia-coli-strain

the steps are, to make it soluble by denature it using Urea or guanidine-HCl, and then refolding it in order to make it active again, Unfortunately this last step are time consuming since many factors affecting their correct folding.It is better to try another methods to correctly express your protein. you can try medium optimization, or environmental culture modification, or try several new plasmids which is help to express your protein into soluble fraction such plasmid contained GroEL/GroES,  

Dear Mohammad,

Here you have my holly Bible of protein recovery from inclusions:

https://www.researchgate.net/publication/13465980_Advances_in_refolding_of_proteins_produced_in_E._coli

Keep fingers cross - Jan

Article Advances in refolding of protein produced in E

Hi Jan, is the full text of this paper available online?

Hi Stefansson, for  that paper you can simply click this link,

http://web.mnstate.edu/provost/Advancesproteinrefolding.pdf

Thanks a lot for your informations. I will try refolding as you suggested.