Inclusion bodies form in E. coli when recombinant protein accumulates faster than the cellular chaperone machinery can fold it. The misfolded protein aggregates into dense, insoluble intracellular particles. Recovery requires: cell disruption → centrifugal isolation of inclusion bodies → solubilisation with chaotropic agents (6–8 M urea or guanidinium chloride) → controlled refolding by dilution or dialysis into native buffer. Despite the extra steps, inclusion bodies offer high-purity protein and protection from proteolysis.