期刊文献+

中国地鼠卵巢细胞培养上清中重组骨形态发生蛋白2的三步纯化法(英文)

A three-step purification method for rhBMP-2 produced in Chinese hamster ovary cells
下载PDF
导出
摘要 骨形态发生蛋白2(BMP-2)是转化生长因子β(TGF-β)超家族的成员,在骨组织工程领域被广泛应用.如今,BMP-2的商业生产所受关注程度与日俱增.这里建立了一种方便且低成本的方法,将人重组骨形态发生蛋白2(rhBMP-2)从中国地鼠卵巢细胞系(CHO)培养上清中纯化出来.该方法由三步组成:肝素亲和层析,Q-Sepharose离子交换层析和SephacrylS-200凝胶过滤,得率约29%.从1L培养上清中能得到约0.26mg的纯度在95%以上的rhBMP-2成熟二聚体,并通过异位成骨实验组织学分析证实了其生物活性.因此,该方法在CHO培养上清中纯化rhBMP-2方面具有潜在的应用价值. Bone morphogenetic protein-2 (BMP-2),a member of the transforming growth factor β superfamily,is widely used in bone tissue engineering. Commercial production of BMP-2 is attracting increasing attention at present time. We have developed a convenient and low-cost purification method of human recombinant BMP-2 (rhBMP-2) from conditioned medium of Chinese hamster ovary cell line (CHO). The three-step procedure with a purification yield of 29% includes heparin affinity chromatography,Q-Sepharose ion exchange chromatography and Sephacryl S-200 gel filtration. About 0.26 mg of purified rhBMP-2 dimer with purity of more than 95% was obtained from 1 L of conditioned medium. It was shown to be biologically active by a histological analysis of ectopic bone formation. In conclusion,this method has the potential application for rhBMP-2 production from CHO.
出处 《南京大学学报(自然科学版)》 CAS CSCD 北大核心 2010年第1期85-91,共7页 Journal of Nanjing University(Natural Science)
基金 The Doctoral Program of Higher Education (20060284045) the Ministry of Education of People’s Republic of China (20060284042,20060284045)
关键词 重组人骨形态发生蛋白2 中国地鼠卵巢细胞系 蛋白纯化 rhBMP-2 Chinese hamster ovary cell line protein purification
  • 相关文献

参考文献17

  • 1Kingsley D M. The TGF-beta superfamily: New members, new receptors, and new genetic tests of function in different organisms. Genes Development, 1994, 8: 133-146.
  • 2Wang E A, Rosen V, D'Alessandro J S, et al. Recombinant human bone morphogenetic protein induces bone formation. Proceedings of National Academy of Science of USA, 1990, 87: 2220- 2224.
  • 3Wozney J M, Rosen V, Celeste A J, et al. No vel regulators of bone formation: Molecular clones and activities. Science, 1988, 242:1528-1534.
  • 4Hogan B L. Bone morphogenetic proteins: Multifunctional regulators of vertebrate development. Genes Development, 1996, 10: 1580- 1594.
  • 5Scheuxer C, Sebald W, Hulsmeyer M. Crystal structure of human bone morphogenetie protein- 2 at 2.7 A resolution. Journal of Molecular Biology, 1999, 287: 103-115.
  • 6Boden S D, Zdeblick T A, Sandhu H S, et al. The use of rhBMP-2 in interbody fusion cages, defnitive evidence of osteoinduction in human: A preliminary report. Spine, 2000, 25: 376-381.
  • 7Kirker-Head C A. Potential applications and delivery strategies for bone morphogenetic proteins. Advanced Drug Delivery Reviews, 2000, 43: 65-92.
  • 8张森林,毛天球.珊瑚/胶原/重组人骨形成蛋白—2复合人工骨修复兔颅骨缺损[J].南京大学学报(自然科学版),1999,35(5):549-553. 被引量:1
  • 9Wang E A, Rosen V, Cordes P, et al. Purification and characterization of other distinct bone-inducing factors. Proceedings of National Academy of Science of USA, 1988, 85: 9484-9488.
  • 10Bessho K, Kusumoto K, Fujimura K, et al. Comparison of recombinant and purified human hone morphogenetic protein. British Journal of Oral Maxillofacial Surgery, 1999, 37: 2-5.

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部