Producing active bone morphogenic protein dimers, useful for inducing growth of bone and cartilage comprises expressing soluble fusion protein monomer followed by refolding and dimerization

摘要

Production of dimeric, biologically active bone morphogenic protein (BMP; I) comprises over expressing and isolating the monomeric form of (I) as a fusion protein in soluble form, in a microbial host. Production of dimeric, biologically active bone morphogenic protein (BMP; I) comprises: (a) over expressing the monomeric form of (I) as a fusion protein (FP), in soluble form, in a microbial host; (b) isolating FP; (c) refolding and dimerizing in renaturing buffer at high protein concentration and without preliminary precipitation; and (d) treating the dimer formed with a protease to form (I). FP comprises the Escherichia coli maltose-binding protein (MBP), a linker that includes a protease cleavage site and BMP.