公开(公告)号：US20150184138A1

公开(公告)日：2015-07-02

申请号：US14448545

申请日：2014-07-31

申请人： Greg Pogue ,  Ernie Hiatt ,  Romy Kandzia ,  Stefan Werner ,  Frank Thieme ,  Tsafrir Mor

发明人： Greg Pogue ,  Ernie Hiatt ,  Romy Kandzia ,  Stefan Werner ,  Frank Thieme ,  Tsafrir Mor

IPC分类号： C12N9/18 ,  A61K38/46

CPC分类号： C12N9/18 ,  A61K38/465 ,  C12Y301/01008

摘要： A new, reliable, easily scalable and reproducible method for the production of recombinant butyrylcholinesterase (rBuChE) is provided. Through the utilization of a plant transfection procedure, various plant strains have been shown to generate effective and scalable amounts of rBuChE under acceptable manufacturing processes to permit reliable levels of such enzymes for desired nerve agent protection requirements (including tetrameric products). As well, such methods in engineered plant lines have shown suitable production of these enzymes in tetramer form with glycan formation and sialyalation (for terminal groups) to allow for optimal potency against organophosphorus agent exposure as well as proper immunogenic response within the plant sources. The overall production method, including the transfection and production within mammalian cells, as well as the process steps involved for such a reliable sourcing platform from plants is thus encompassed within the invention.

摘要翻译： 提供了一种用于生产重组丁酰胆碱酯酶（rBuChE）的新的，可靠的，易于扩展和重现的方法。 通过利用植物转染方法，已经显示各种植物菌株在可接受的制造过程中产生有效和可缩放量的rBuChE，以允许可靠水平的这些酶用于所需的神经药物保护要求（包括四聚物产品）。 同样，工程化植物品系中的这些方法已经显示出以四聚体形式合适地生产这些酶，其中聚糖形成和sialyalation（用于末端基团）以允许针对有机磷剂暴露的最佳效力以及在植物来源内的适当的免疫原性反应。 因此，包括哺乳动物细胞内的转染和生产的总体生产方法以及从植物获得这种可靠的采购平台所涉及的工艺步骤。