Laboratori de Fisiologia Vegetal, Facultat de Farmacia, Universitat de Barcelona, Av. Joan XXIII sn, 08028 Barcelona, Spain.
Department of Biotechnology, Institute of Science, High Technology and Environmental Science, Graduate University of Advanced Tecnology, P.O. Box 76315-117, Kerman, Iran.
Plant Physiol Biochem. 2017 Sep;118:130-137. doi: 10.1016/j.plaphy.2017.06.013. Epub 2017 Jun 13.
Transplastomic plants are a system of choice for the mass production of biopharmaceuticals due to the polyploidy of the plastid genome and the low risk of pollen-mediated outcrossing because of maternal inheritance. However, as field-grown plants, they can suffer contamination by agrochemicals and fertilizers, as well as fluctuations in yield due to climatic changes and infections. Tissue-type plasminogen activator (tPA), a protein used to treat heart attacks, converts plasminogen into plasmine, which digests fibrin and induces the dissolution of fibrin clots. Recently, we obtained transplastomic tobacco plants carrying the K2S gene encoding truncated human tPA (reteplase) with improved biological activity, and confirmed the presence of the target protein in the transgenic plant leaves. Considering the advantages of plant cell cultures for biopharmaceutical production, we established a cell line derived from the K2S tobacco plants. The active form of reteplase was quantified in cultures grown in light or darkness, with production 3-fold higher in light.
转叶绿体植物由于质体基因组的多倍性和母系遗传导致花粉介导的杂交的风险低，是生物制药大规模生产的首选系统。然而，作为田间生长的植物，它们可能会受到农用化学品和肥料的污染，以及由于气候变化和感染导致产量波动。组织型纤溶酶原激活剂（tPA）是一种用于治疗心脏病发作的蛋白质，它将纤溶酶原转化为纤溶酶，分解纤维蛋白并诱导纤维蛋白凝块溶解。最近，我们获得了携带编码截断人 tPA（瑞替普酶）的 K2S 基因的转叶绿体烟草植物，该基因具有改善的生物学活性，并在转基因植物叶片中证实了目标蛋白的存在。考虑到植物细胞培养在生物制药生产中的优势，我们建立了一种源自 K2S 烟草植物的细胞系。在光照或黑暗中培养的细胞中定量了瑞替普酶的活性形式，光照下的产量增加了 3 倍。
