title: [Production of the recombinant human bone morphogenetic protein-2 in Escherichia coli and testing of its biological activity in vitro and in vivo].
abstract: Bone morphogenetic protein-2 (rhBMP-2) represents the osteoinductive protein factor which plays a dominant role in growth and regeneration of a bone tissue. In clinical practice the bone grafting materials on the basis of rhBMP-2 are widely applied; the Russian analogues of similar materials are not produced. The fragment of the bmp2gene coding for a mature protein was cloned in Escherichia coli. The effective overproducing strain of rhBMP-2 was created on a basis of the E. coli BL21 (DE3). The rhBMP-2 production was about 25% of total cell protein. The biologically active dimeric form of rhBMP-2 was obtained by isolation and purification of protein from inclusion bodies with subsequent refolding. The rhBMP-2 sample with more than 80% of the dimeric form was obtained, which is able to interact with specific antibodies to BMP-2. Biological activity of the received rhBMP-2 samples was shown in the in vitro experiments by induction of alkaline phosphatase synthesis in C2C12 and C3H10T1/2 cell cultures. On model of the ectopic osteogenesis it was shown that received rhBMP-2 possesses biological activity in vivo, causing tissue calcification in the place of an injection. The protein activity in vivo depends on way of protein introduction and characteristics of protein sample: rhBMP-2 may be introduced in an acid or basic buffer solution, with or without the carrier. The offered method of rhBMP-2 isolation and purification results in increasing common protein yield as well as the maintenance of biologically active dimeric form in comparison with the analogues described in the literature.
authors.forename: N E
authors.initials: NE
authors.lastname: Sharapova
authors.forename: A P
authors.initials: AP
authors.lastname: Kotnova
authors.forename: Z M
authors.initials: ZM
authors.lastname: Galushkina
authors.forename: N V
authors.initials: NV
authors.lastname: Lavrova
authors.forename: N N
authors.initials: NN
authors.lastname: Poletaeva
authors.forename: A E
authors.initials: AE
authors.lastname: Tukhvatulin
authors.forename: A E
authors.initials: AE
authors.lastname: Tukhvatullin
authors.forename: A S
authors.initials: AS
authors.lastname: Semikhin
authors.forename: A V
authors.initials: AV
authors.lastname: Gromov
authors.forename: L A
authors.initials: LA
authors.lastname: Soboleva
authors.forename: A S
authors.initials: AS
authors.lastname: Ershova
authors.forename: V V
authors.initials: VV
authors.lastname: Zaĭtsev
authors.forename: O V
authors.initials: OV
authors.lastname: Sergienko
authors.forename: V G
authors.initials: VG
authors.lastname: Lunin
authors.forename: A S
authors.initials: AS
authors.lastname: Kariagina
mesh_terms: D000595:Amino Acid Sequence; D055396:Bone Morphogenetic Protein 2 / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D005091:Exons / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D011994:Recombinant Proteins