title: [Prokaryotic expression of recombinant human bone morphogenetic protein-2 and preparation of monoclonal antibodies].
abstract: OBJECTIVE
  To express and purify the recombinant human bone morphogenetic protein-2 mature peptide (rhBMP-2m) in prokaryotic system and to develop highly-specific monoclonal antibodies.
  
  
  METHODS
  An engineered E. coli strain expressing rhBMP-2m was fermented. The bacterial cells were firstly lysed and then the rhBMP-2m inclusion bodies were isolated by centrifugation. After the inclusion bodies had been solubilized by high-concentration denaturing agents, denatured rhBMP-2m was purified by cation ion-exchange chromatography. Biologically active rhBMP-2m was obtained by refolding of purified denatured rhBMP-2m through direct dilution. The refolded rhBMP-2m was used to immunize Balb/c mice to develop anti-rhBMP-2m monoclonal antibodies using classic hybridoma technique.
  
  
  RESULTS
  rhBMP-2m with a purity greater than 95% was obtained on reduced SDS-PAGE. The refolded rhBMP-2m was measured to be bioactive by the induction of alkaline phosphatase activity in MC3T3-E1 cells. Two hybridoma cell lines that stably secreted anti-rhBMP-2m antibody were developed from the immunized mice.
  
  
  CONCLUSIONS
  Bioactive rhBMP-2m protein and its monoclonal antibodies were successfully prepared, which will provides a solid base for future studies on rhBMP-2.
authors.affiliation: Cell Culture and Center, CAMS and PUMC, Beijing 100005, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ning-Ning
authors.initials: NN
authors.lastname: Ma
authors.forename: Liang-Zhi
authors.initials: LZ
authors.lastname: Xie
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000276:immunology; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D019485:Bone Morphogenetic Proteins