METHOD OF PRODUCTION OF PROTEIN-MINERAL COMPOSITION CONTAINING RECOMBINANT PROTEIN Collbd-BMP-2

摘要

FIELD: biotechnology.SUBSTANCE: first a plasmid pCollbd-BMP-2 is produced. Then, said plasmid is inserted into the strain Escherichia coli M15/pREP4. Then the recombinant protein Collbd-BMP-2 is prepared by culturing the cells of a strain Escherichia coli Ml 5 [pREP4, pCollbd-BMP-2], the induction of protein Collbd-BMP-2 synthesis, the cell destruction, obtaining the supernatant containing the protein, immobilisation of protein by adding to the supernatant of suspension of collagen-containing sorbent, incubation, washing the sorbent with a related protein, the subsequent elution of protein from the sorbent and dialysis. Then, in a laminar flow unit two sterile polypropylene tubes are prepared and the hydroxyapatite suspension is placed in them, a solution of connective-tissue collagen or xenogenic bone collagen in the form of crumbs is added, mixed on a shaker to obtain a homogeneous suspension. Then 10% (by weight) of gelatin microspheres containing recombinant growth factor of bone tissue Collbd-BMP-2 is added, stirred on the shaker. The resulting composition is sterile pasta of white, beige, grayish or yellowish colour of uniform consistency, contains 5-20% of synthetic nanostructured hydroxyapatite, 0.1-15% of type I collagen and 5-10% of a prolonged form of recombinant growth factor of bone tissue Coilbd-BMP 2 and has osteoconductive and osteoinductive properties. The composition according to the present invention can be used to fill bone defects and be applied on the implantable materials.EFFECT: invention can be used in traumatology, orthopedics, maxillofacial surgery, dentistry for treatment of diseases and injuries of human osseous system as an active biodegradable material, for bone tissue regeneration, osteoinductive biological coating of metal prosthetic implants of bone tissue.4 ex