Human bone morphogenetic protein-2 (hBMP-2) is a potent growth and differentiation factor for bone induction and regeneration. Recombinant hBMP-2 (rhBMP-2) was cloned and expressed as a soluble protein using E. coli-based expression system. A full-length gene encoding mature hBMP-2 was amplified by RT-PCR, cloned into an expression vector and expressed using SHuffle E. coli cells. The rhBMP-2 was successfully expressed as a soluble protein under the control of the lacUV5 and protein A promoters by IPTG induction. The rhBMP-2 fused with ZZ domain at its N-terminus was successively purified with a single step by using IgG Sepharose 6 fast flow affinity chromatography. Analysis of the purified protein on SDS-PAGE, Western blot analysis and LC-MS/MS, verified that the purified protein was rhBMP-2. The biological activity testing on hFOB 1.19 showed that rhBMP-2 had the ability to significantly induce cell proliferation in a dose dependent manner. ALP staining and activity assay also increased after rhBMP-2