Expression, purification and characterization of bone morphogenetic protein (BMP-2) for application in composites biomaterials with poly(E-caprolactone)

Abstract

The company Biosintesis already obtained grants from FAPESP via the PIPE program (phase 1 and 2) and PAPPE-PIPE 3, in addition to the investments done by the same company. A new PIPE project was recently submitted for obtaining and characterizing membranes made with porose electrospinned nanofibers of poly (µ-caprolactone) (PCL), associated to functional bioactive products for the construction of scaffolds, as biological substitutes for tissue engineering. This considers the formation of composites of PCL with bioactive agents, one of which will be human Bone Morphogenetic Protein (BMP)-2, which has an extremely important role in bone regeneration and repair and that, in the submitted project, will necessarily be imported. The present project is therefore considering the proprietary synthesis of the same BMP-2, done by Biosintesis in collaboration with the Biotechnology Center of IPEN-CNEN/São Paulo, which already has all technology and experience that are necessary for this type of development. The periplasmic expression in genetically modified E.coli will provide an already perfectly folded and bioactive protein, avoiding costly and time-consuming solubilization and refolding of cytoplasmic inclusion bodies, which will compensate for the well-known lower yields of periplasmic expression. The final product will be extensively purified and obtained in a stable lyophilized form that will be characterized via physical chemical and biological techniques, comparing it with an International Reference Preparation of CHO-derived BMP-2. A comparison will be made by setting up also a composite of PCL with human growth hormone (hGH), a protein that has already shown good osteogenic activities and that has already been patented by the same Biotechnology Center of IPEN. Besides being BMP-2 in itself a recombinant protein with very high aggregated value and commercially interesting, its synthesis and patent will approximate Biosintesis to the construction of biological scaffolds totally based on Brazilian technology that shall be able to compete on the global market. (AU)