Rattanapisit Kaewta, Srifa Suchada, Kaewpungsup Pornjira, Pavasant Prasit, Phoolcharoen Waranyoo
Research Unit for Plant-Produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand.
Research Unit of Mineralized Tissue, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Biotechnol Rep (Amst). 2019 Feb 8;21:e00312. doi: 10.1016/j.btre.2019.e00312. eCollection 2019 Mar.
Osteopontin (OPN) plays an important role in the bone regeneration process. Previous investigation showed that recombinant human OPN was able to express in leaves and induced the osteogenic related genes. Nevertheless, the purification of OPN from plant proteins with Ni affinity chromatography was still not effective enough. To improve the quality of protein expression and purification in plants, we constructed an Fc-based form of OPN. The complete OPN protein was fused to the human IgG1 Fc domain. Here, we showed that the plant-produced OPN-Fc increases the protein expression level and facilitates the purification of the recombinant protein. Our result showed that the plant-produced OPN-Fc can stimulate the expression of osteogenic related genes such as DMP1, OSX, and Wnt3a and also the calcium deposition in hPDL cells. These findings suggest that the plant-produced OPN-Fc has potential application in tissue engineering in the future.
骨桥蛋白（OPN）在骨再生过程中发挥着重要作用。先前的研究表明，重组人OPN能够在叶片中表达并诱导成骨相关基因。然而，利用镍亲和层析从植物蛋白中纯化OPN仍然不够有效。为了提高植物中蛋白质表达和纯化的质量，我们构建了一种基于Fc的OPN形式。完整的OPN蛋白与人IgG1 Fc结构域融合。在此，我们表明植物产生的OPN-Fc提高了蛋白质表达水平并促进了重组蛋白的纯化。我们的结果表明，植物产生的OPN-Fc可以刺激成骨相关基因如DMP1、OSX和Wnt3a的表达，以及人牙周膜细胞中的钙沉积。这些发现表明，植物产生的OPN-Fc在未来的组织工程中具有潜在的应用价值。
