One-shot method for purification of multiple natural amelogenin isoforms
来源期刊：JOURNAL OF MATERIALS SCIENCE TECHNOLOG2018年第3期
论文作者：Chunbao Wang Jing Zhang Fan Yang Chang Du
文章页码：481 - 487
摘 要：Amelogenin isoforms constitute the predominant component in the enamel matrix and each amelogenin isoform executes unique role in the enamel biomineralization process. Enamel matrix derivative enriching amelogenin isoforms have also bioactive property for tissue regeneration. Despite the development of recombinant protein technology that has greatly forwarded the understanding of amelogenin properties,substantial evidences have revealed biochemical and functional difference between natural amelogenins and their recombinant form. To facilitate the study of enamel formation mechanism, more facile methodology to purify multiple natural amelogenin isoforms is pursued. Here we developed an effective one-shot method via reverse phase high-performance liquid chromatography（RP-HPLC） to purify various amelogenin isoforms from pig-derived amelogenin complex. A thorough process of chromatographic condition establishment including sample analysis on analytical scale and chromatographic condition design on preparative scale was described. Three representative amelogenin isoforms（TRAP, P148, P173） were isolated in one step and their purity was confirmed by matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS） and high resolution mass spectrometry.
Chunbao Wang1,2，Jing Zhang1,2，Fan Yang1,2，Chang Du1,2,3
1. School of Materials Science and Engineering, South China University of Technology2. National Engineering Research Center for Tissue Restoration & Reconstruction, South China University of Technology3. Key Laboratory of Biomedical Materials Science and Engineering, Ministry of Education,Guangzhou
摘 要：Amelogenin isoforms constitute the predominant component in the enamel matrix and each amelogenin isoform executes unique role in the enamel biomineralization process. Enamel matrix derivative enriching amelogenin isoforms have also bioactive property for tissue regeneration. Despite the development of recombinant protein technology that has greatly forwarded the understanding of amelogenin properties,substantial evidences have revealed biochemical and functional difference between natural amelogenins and their recombinant form. To facilitate the study of enamel formation mechanism, more facile methodology to purify multiple natural amelogenin isoforms is pursued. Here we developed an effective one-shot method via reverse phase high-performance liquid chromatography（RP-HPLC） to purify various amelogenin isoforms from pig-derived amelogenin complex. A thorough process of chromatographic condition establishment including sample analysis on analytical scale and chromatographic condition design on preparative scale was described. Three representative amelogenin isoforms（TRAP, P148, P173） were isolated in one step and their purity was confirmed by matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS） and high resolution mass spectrometry.
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