Over-expression of proteins in prokaryotes often leads to the production of insoluble aggregates of misfolded proteins in inclusion bodies. These misfolded proteins lack their desired natural functionality. Sometimes considered a nuisance, the formation of inclusion bodies has also some advantages, such as high enrichment of desired protein and protection against protease activities. Regardless the fact that the method for purification of inclusion bodies should be evaluated to each one different recombinant protein, usually the methods include mechanical or chemical cell disruption using chaotropic agents like guanidine salt or urea.

Three recombinant proteins that show different biochemical characteristics were purified from inclusion bodies using a basic method that comprised of two principal steps: cell aggregate chemical disruption and inclusion bodies washing. By including slight variations a comparison was achieved in order to define how such modification could contribute or affect the performance of our inclusion body purification method.