METHOD FOR PRODUCING A BONE REGENERATION AGENT CONTAINING RECOMBINANT BMP-2 PROTEIN AND ITS USE Russian patent published in 2022 - IPC A61K35/32 C12N15/12 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. Described is a method for obtaining recombinant human bone morphogenetic protein 2 (bone morphogenetic protein 2, BMP-2) by transforming E. coli cells with the pET-s-tag-BMP-2 plasmid followed by chromatographic purification of the protein and obtaining an agent containing such a protein by immobilization the specified protein on the carrier. The carrier is a mineral bone matrix or demineralized bone matrix (DBM). The invention also relates to a tool for bone regeneration, including a carrier immobilized on it with the recombinant BMP-2 protein. The tool is made in the form of granules, blocks or membranes. The invention also relates to the use of said agent as an osteoplastic material for replenishing the anatomical integrity of bone tissue. The effect is achieved by increasing the solubility and formation of the native BMP-2 conformation, which are the result of the presence of an additional protein domain in the protein and optimization of purification and refolding conditions. More precisely, the effect is achieved by including an additional protein domain - s-tag - a 15-mer oligopeptide from ribonuclease A of the pancreas of a mammal, for example, a bull, into the composition of the recombinant BMP-2. It was found that the final properties of the composition, including a mineral bone matrix or demineralized bone matrix and BMP-2 protein, expressed in increased osteoinductivity and storage stability, are determined by the properties of BMP-2, namely, increased BMP-2 activity and its solubility, due to introduction of an additional “s-tag” domain into the protein structure. Features of protein immobilization on the carrier provide clinically substantiated kinetics of the release of a certain amount of protein from the carrier for two weeks after implantation. In addition, due to immobilization, a prolonged biological effect and an increase in osteoinductivity in the area of ​​bone tissue regeneration are provided, which in turn will reduce the duration of treatment in days, increase the volume of formed bone tissue, increase the predictability of achieving a clinical result of treatment, such as bone formation and recovery of bone defect.

EFFECT: ensuring stable parameters and high controlled osteoinductive properties of the agent, independent of changes in storage conditions, the possibility of obtaining the agent in a physical form convenient for use and storage.

12 cl, 7 dwg, 10 ex