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    • 5. 发明申请
    • IMPROVED BACTERIAL OUTER MEMBRANE VESICLES
    • 改进的细菌外膜VESICLES
    • WO2004019977A2
    • 2004-03-11
    • PCT/IB2003/004293
    • 2003-09-01
    • CHIRON SRLPIZZA, MariagraziaSERRUTO, DavideRAPPUOLI, Rini
    • PIZZA, MariagraziaSERRUTO, DavideRAPPUOLI, Rini
    • A61K39/095
    • A61K39/095A61K2039/55555C07K14/22C12N1/06C12N1/20C12N13/00
    • Existing methods of meningococcal OMV preparation involve the use of detergent during disruption of the bacterial membrane. According to the invention, membrane disruption is performed substantially in the absence of detergent. The resulting OMVs which retain important bacterial immunogenic components, particularly (i) the protective NspA surface protein, (ü) protein NMB2132 and (iii) protein NMB 1870.A Typical process involves the following steps: (a) treating bacterial cells in the substantial absence of detergent; (b) centrifuging the composition from step (a) to separate the outer membrane vesicles from treated cells and cell debris, and collecting the supernatant; (c) performing a high speed centrifugation of the supernatant from step (b) and collecting the outer membrane vesicles in a pellet; (d) re-dispersing the pellet from step (c) in a buffer; (e) performing a second high speed centrifugation in accordance with step (c), collecting the outer membrane vesicles in a pellet; (f) re-dispersing the pellet from step (e) in an aqueous medium.
    • 脑膜炎球菌OMV制剂的现有方法涉及在细菌膜破坏期间使用洗涤剂。 根据本发明,膜破坏基本上在没有洗涤剂的情况下进行。 所产生的OMV保留重要的细菌免疫原性成分,特别是(i)保护性NspA表面蛋白(ü)蛋白NMB2132和(iii)蛋白NMB1870.A典型的方法包括以下步骤:(a) 不含洗涤剂; (b)将来自步骤(a)的组合物离心以将外膜囊泡与经处理的细胞和细胞碎片分离,并收集上清液; (c)对来自步骤(b)的上清液进行高速离心并将颗粒中的外膜囊泡收集; (d)将来自步骤(c)的沉淀物重新分散在缓冲液中; (e)根据步骤(c)进行第二高速离心,将外膜囊泡收集在颗粒中; (f)将来自步骤(e)的沉淀物重新分散在水性介质中。
    • 8. 发明申请
    • IMPROVED BACTERIAL OUTER MEMBRANE VESICLES
    • 改进的细菌外膜膜
    • WO2004019977A3
    • 2004-07-15
    • PCT/IB0304293
    • 2003-09-01
    • CHIRON SRLPIZZA MARIAGRAZIASERRUTO DAVIDERAPPUOLI RINI
    • PIZZA MARIAGRAZIASERRUTO DAVIDERAPPUOLI RINI
    • A61K9/00A61K39/095C12N1/06A61K39/104A61K39/112
    • A61K39/095A61K2039/55555C07K14/22C12N1/06C12N1/20C12N13/00
    • Existing methods of meningococcal OMV preparation involve the use of detergent during disruption of the bacterial membrane. According to the invention, membrane disruption is performed substantially in the absence of detergent. The resulting OMVs which retain important bacterial immunogenic components, particularly (i) the protective NspA surface protein, (ü) protein NMB2132 and (iii) protein NMB 1870.A Typical process involves the following steps: (a) treating bacterial cells in the substantial absence of detergent; (b) centrifuging the composition from step (a) to separate the outer membrane vesicles from treated cells and cell debris, and collecting the supernatant; (c) performing a high speed centrifugation of the supernatant from step (b) and collecting the outer membrane vesicles in a pellet; (d) re-dispersing the pellet from step (c) in a buffer; (e) performing a second high speed centrifugation in accordance with step (c), collecting the outer membrane vesicles in a pellet; (f) re-dispersing the pellet from step (e) in an aqueous medium.
    • 现有的脑膜炎球菌OMV制备方法涉及在破坏细菌膜期间使用去污剂。 根据本发明,膜破裂基本上在不含洗涤剂的情况下进行。 (i)保护性NspA表面蛋白质,(ü)蛋白质NMB2132和(iii)蛋白质NMB 1870.典型的方法包括以下步骤:(a)处理大量的细菌免疫原性组分 没有洗涤剂; (b)离心步骤(a)的组合物以从处理的细胞和细胞碎片中分离外膜囊泡,并收集上清液; (c)对来自步骤(b)的上清液进行高速离心并将外膜囊泡收集在沉淀中; (d)将来自步骤(c)的颗粒重新分散在缓冲液中; (e)根据步骤(c)进行第二次高速离心,收集沉淀中的外膜囊泡; (f)将来自步骤(e)的粒料再分散在含水介质中。