• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

通过物理化学、免疫学和生物学分析对人骨形态发生蛋白-2（hBMP-2）进行表征。

Human bone morphogenetic protein-2 (hBMP-2) characterization by physical-chemical, immunological and biological assays.

作者信息

Suzuki Miriam Fussae, Oliveira João Ezequiel, Damiani Renata, Lima Eliana Rosa, Amaral Kleicy Cavalcante, Santos Anderson Maikon de Souza, Magalhães Geraldo Santana, Faverani Leonardo Perez, Pereira Luis Antonio Violin Dias, Silva Fabiana Medeiros, Bartolini Paolo

机构信息

Biotechnology Center, Instituto de Pesquisas Energéticas e Nucleares, IPEN-CNEN/SP, Avenida Prof. Lineu Prestes 2242, Cidade Universitária, São Paulo, SP, 05508-000, Brazil.

Biosintesis P&D, São Paulo, SP, Brazil.

出版信息

AMB Express. 2020 Feb 17;10(1):34. doi: 10.1186/s13568-020-0964-5.

DOI:10.1186/s13568-020-0964-5
PMID:32067115
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7026339/
Abstract

Commercially available preparations of methionyl-human BMP-2 and CHO-derived hBMP-2, which belongs to the transforming growth factor β (TGF-β) superfamily, were used for a complete characterization. This protein is an extremely efficient osteoinductor that plays an important role during bone regeneration and embryonic development. Characterization was carried out via SDS-PAGE and Western blotting, followed by reversed-phase HPLC, size-exclusion HPLC and MALDI-TOF-MS. The classical in vitro bioassay, based on the induction of alkaline phosphatase activity in C2C12 cells, confirmed that hBMP-2 biological activity is mostly related to the dimeric form, being ~ 4-fold higher for the CHO-derived glycosylated form when compared with the E. coli counterpart. The E. coli-derived met-hBMP-2 has shown, by MALDI-TOF-MS, a large presence of the bioactive dimer. A more complex molecular mass (MM) distribution was found for the CHO-derived product, whose exact MM has never been reported because of its variable glycosylation. A method based on RP-HPLC was set up, allowing a quantitative and qualitative hBMP-2 determination even directly on ongoing culture media. Considering that hBMP-2 is highly unstable, presenting moreover an extremely high aggregate value, we believe that these data pave the way to a necessary characterization of this important factor when synthesized by DNA recombinant techniques in different types of hosts.

摘要

使用市售的甲硫氨酰人骨形态发生蛋白-2（methionyl-human BMP-2）制剂和源自中国仓鼠卵巢细胞（CHO）的人骨形态发生蛋白-2（hBMP-2）进行全面表征，hBMP-2属于转化生长因子β（TGF-β）超家族。这种蛋白质是一种极其有效的骨诱导剂，在骨再生和胚胎发育过程中发挥着重要作用。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹法（Western blotting）进行表征，随后进行反相高效液相色谱（RP-HPLC）、尺寸排阻高效液相色谱和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）。基于C2C12细胞中碱性磷酸酶活性诱导的经典体外生物测定法证实，hBMP-2的生物活性主要与二聚体形式相关，与大肠杆菌来源的对应物相比，源自CHO的糖基化形式的活性高约4倍。通过MALDI-TOF-MS分析，大肠杆菌来源的甲硫氨酰-hBMP-2显示出大量生物活性二聚体。发现源自CHO的产品具有更复杂的分子量（MM）分布，由于其糖基化可变，其确切的MM从未被报道过。建立了一种基于RP-HPLC的方法，甚至可以直接对正在培养的培养基进行hBMP-2的定量和定性测定。考虑到hBMP-2高度不稳定，且聚集值极高，我们认为这些数据为通过DNA重组技术在不同类型宿主中合成该重要因子时进行必要的表征铺平了道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/dce78a46ae62/13568_2020_964_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/f850be2ebb0c/13568_2020_964_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/c4506b972348/13568_2020_964_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/17779fc87c55/13568_2020_964_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/13597d1ce929/13568_2020_964_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/a1f88ec334f3/13568_2020_964_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/dce78a46ae62/13568_2020_964_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/f850be2ebb0c/13568_2020_964_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/c4506b972348/13568_2020_964_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/17779fc87c55/13568_2020_964_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/13597d1ce929/13568_2020_964_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/a1f88ec334f3/13568_2020_964_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8e3/7026339/dce78a46ae62/13568_2020_964_Fig6_HTML.jpg

相似文献

1
Human bone morphogenetic protein-2 (hBMP-2) characterization by physical-chemical, immunological and biological assays.通过物理化学、免疫学和生物学分析对人骨形态发生蛋白-2（hBMP-2）进行表征。
AMB Express. 2020 Feb 17;10(1):34. doi: 10.1186/s13568-020-0964-5.
2
Synthesis of Human Bone Morphogenetic Protein-2 (hBMP-2) in Periplasmic Space: Its Characterization and Preclinical Testing.人骨形态发生蛋白-2（hBMP-2）在周质空间中的合成：其表征和临床前测试。
Cells. 2021 Dec 14;10(12):3525. doi: 10.3390/cells10123525.
3
[Expression of recombinant human BMP-6 in Escherichia coli and its purification and bioassay in vitro].重组人骨形态发生蛋白-6在大肠杆菌中的表达及其体外纯化与生物学活性测定
Sheng Wu Gong Cheng Xue Bao. 2003 Sep;19(5):556-60.
4
[Expression, characterization and biological activity analysis of recombinant human bone morphogenetic protein 2 in CHO cells].重组人骨形态发生蛋白2在CHO细胞中的表达、鉴定及生物学活性分析
Sheng Wu Gong Cheng Xue Bao. 2006 Nov;22(6):968-72.
5
Adenovirus mediated BMP-13 gene transfer induces chondrogenic differentiation of murine mesenchymal progenitor cells.腺病毒介导的BMP - 13基因转移诱导小鼠间充质祖细胞的软骨形成分化。
J Bone Miner Res. 2004 Jan;19(1):111-22. doi: 10.1359/jbmr.2004.19.1.111.
6
[Bone-inducing activity of human bone morphogenetic protein-2 102 peptide].[人骨形态发生蛋白-2 102肽的骨诱导活性]
Sheng Wu Gong Cheng Xue Bao. 2001 Nov;17(6):678-82.
7
[Human bone morphogenetic protein-2: recombinant expression in E. coli].[人骨形态发生蛋白-2：在大肠杆菌中的重组表达]
Zhonghua Yi Xue Za Zhi. 1998 Apr;78(4):308-10.
8
Two-step chromatographic purification of recombinant human thyrotrophin and its immunological, biological, physico-chemical and mass spectral characterization.重组人促甲状腺素的两步色谱纯化及其免疫学、生物学、物理化学和质谱表征
J Chromatogr A. 2005 Jan 7;1062(1):103-12. doi: 10.1016/j.chroma.2004.10.084.
9
Chaperone-mediated production of active homodimer human bone morphogenetic protein - 2 in E. coli.伴护介导的大肠杆菌中活性同型二聚体人骨形态发生蛋白-2的生产。
Protein Expr Purif. 2023 Jun;206:106245. doi: 10.1016/j.pep.2023.106245. Epub 2023 Feb 16.
10
Expression and purification of recombinant bone morphogenetic protein-2 in E.coli.重组骨形态发生蛋白-2在大肠杆菌中的表达与纯化
Di Yi Jun Yi Da Xue Xue Bao. 2002 May;22(5):413-6.

引用本文的文献

1
Enhancement of In Vivo Bone Regeneration by the Carbohydrate Derivative DP2.碳水化合物衍生物DP2对体内骨再生的促进作用
Pharmaceuticals (Basel). 2025 Feb 5;18(2):215. doi: 10.3390/ph18020215.
2
DP2, a Carbohydrate Derivative, Enhances In Vitro Osteoblast Mineralisation.DP2，一种碳水化合物衍生物，可增强体外成骨细胞矿化。
Pharmaceuticals (Basel). 2023 Oct 24;16(11):1512. doi: 10.3390/ph16111512.
3
Assessment of Escherichia coli-derived Recombinant Human Bone Morphogenic Protein-2 on Fertilization and Early Embryonic Development in Rats.

本文引用的文献

1
BMP-2 showed comparable osteoinductivity with Chinese hamster ovary derived BMP-2 with demineralized bone matrix as carrier.以脱矿骨基质为载体时，BMP - 2与中国仓鼠卵巢细胞来源的BMP - 2表现出相当的骨诱导活性。
Growth Factors. 2019 Apr;37(1-2):85-94. doi: 10.1080/08977194.2019.1596905. Epub 2019 Apr 4.
2
Determination of recombinant Interferon-α2 in E. coli periplasmic extracts by reversed-phase high-performance liquid chromatography.采用反相高效液相色谱法测定大肠杆菌周质提取物中的重组干扰素-α2 。
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Jan 1;1072:193-198. doi: 10.1016/j.jchromb.2017.11.023. Epub 2017 Nov 21.
3
评估大肠杆菌来源的重组人骨形态发生蛋白-2 对大鼠受精和早期胚胎发育的影响。
Pharm Res. 2023 Jun;40(6):1569-1576. doi: 10.1007/s11095-023-03514-z. Epub 2023 Apr 20.
4
Regulatory of miRNAs in tri-lineage differentiation of C3H10T1/2.miRNAs 在 C3H10T1/2 三系分化中的调控作用。
Stem Cell Res Ther. 2022 Nov 22;13(1):521. doi: 10.1186/s13287-022-03205-3.
5
Modulation of osteogenic differentiation by Escherichia coli-derived recombinant bone morphogenetic protein-2.大肠杆菌衍生的重组骨形态发生蛋白-2对成骨分化的调节作用
AMB Express. 2022 Aug 10;12(1):106. doi: 10.1186/s13568-022-01443-5.
6
Synthesis of Human Bone Morphogenetic Protein-2 (hBMP-2) in Periplasmic Space: Its Characterization and Preclinical Testing.人骨形态发生蛋白-2（hBMP-2）在周质空间中的合成：其表征和临床前测试。
Cells. 2021 Dec 14;10(12):3525. doi: 10.3390/cells10123525.
Comparison of the effects of recombinant human bone morphogenetic protein-2 and -9 on bone formation in rat calvarial critical-size defects.
重组人骨形态发生蛋白-2 和 -9 对大鼠颅骨临界尺寸缺损中成骨作用的比较。
Clin Oral Investig. 2017 Dec;21(9):2671-2679. doi: 10.1007/s00784-017-2069-3. Epub 2017 Feb 14.
4
Histological analysis and gene expression profile following augmentation of the anterior maxilla using rhBMP-2/ACS versus autogenous bone graft.使用重组人骨形态发生蛋白-2/可吸收胶原海绵与自体骨移植对上颌前部进行增容后的组织学分析及基因表达谱
J Clin Periodontol. 2016 Dec;43(12):1200-1207. doi: 10.1111/jcpe.12601. Epub 2016 Oct 3.
5
Bone morphogenetic protein in spine surgery: current and future uses.脊柱外科中的骨形态发生蛋白：当前和未来的应用。
J Am Acad Orthop Surg. 2012 Sep;20(9):547-52. doi: 10.5435/JAAOS-20-09-547.
6
Comparative study of Chinese hamster ovary cell versus Escherichia coli-derived bone morphogenetic protein-2 using the critical-size supraalveolar peri-implant defect model.采用临界尺寸牙槽嵴顶周围种植体缺损模型比较中国仓鼠卵巢细胞与大肠杆菌来源骨形成蛋白-2。
J Periodontol. 2013 Mar;84(3):415-22. doi: 10.1902/jop.2012.110369. Epub 2012 May 21.
7
Effect of Escherichia coli-produced recombinant human bone morphogenetic protein 2 on the regeneration of canine segmental ulnar defects.大肠杆菌表达的重组人骨形态发生蛋白 2 对犬桡骨节段性缺损再生的影响。
J Bone Miner Metab. 2012 Jul;30(4):388-99. doi: 10.1007/s00774-011-0329-x. Epub 2011 Nov 2.
8
Bone morphogenetic proteins for periodontal and alveolar indications; biological observations - clinical implications.用于牙周和牙槽适应症的骨形态发生蛋白；生物学观察 - 临床意义
Orthod Craniofac Res. 2009 Aug;12(3):263-70. doi: 10.1111/j.1601-6343.2009.01461.x.
9
Osteoinductive capacity and heat stability of recombinant human bone morphogenetic protein-2 produced by Escherichia coli and dimerized by biochemical processing.由大肠杆菌产生并经生化处理二聚化的重组人骨形态发生蛋白-2的骨诱导能力和热稳定性。
J Bone Miner Metab. 2009;27(3):355-63. doi: 10.1007/s00774-009-0040-3. Epub 2009 Feb 21.
10
Expression, purification, and renaturation of bone morphogenetic protein-2 from Escherichia coli.从大肠杆菌中表达、纯化及复性骨形态发生蛋白-2
Protein Expr Purif. 2006 Apr;46(2):374-8. doi: 10.1016/j.pep.2005.09.025. Epub 2005 Oct 27.