Hi everyone,

I have a very troubling question with the project I'm working on.

So just a brief overview: we want to produce a eukaryotic recombinantly, the host is E. coli. Production is done is 10 L bioreactor scale. The protein is produced in inclusion bodies. Last time I did the purification using IMAC on ÄKTA using buffer containing urea. As far as I know, inclusion bodies can only be solubilized in urea or Gua HCl. Also, this protein has a very low solubility in water which is why I have to use other methods for the purification.

However, now we want to do other methods without the usage of urea (or Gua HCl). Does anyone know if there is any way to solubilize protein inside inclusion bodies without urea?

Or by any chance, how to increase the solubility of casein? I have been struggling with this and could not find other suitable methods.

I even tried the simple ammonium sulfate precipitation and of course it didn't work.

Could someone please provide me insights on which method or modification I have to do for the purification of this protein. It doesn't have to be very clean or crystal structure.

The goal is to make it more soluble in water or polar solvents.

Thank you.

Kind regards,

Lieke