公开(公告)号：US20070249019A1

公开(公告)日：2007-10-25

申请号：US11788071

申请日：2007-04-19

Applicant: Yun Kang ,  Mark William Cutler ,  Amadou Affrey Ouattara ,  Kristen Elissa Syvertsen

Inventor： Yun Kang ,  Mark William Cutler ,  Amadou Affrey Ouattara ,  Kristen Elissa Syvertsen

IPC: C12Q1/68 ,  C12P21/06

CPC classification number: C12N7/00 ,  A61K39/00 ,  A61K48/00 ,  A61K2039/5252 ,  A61K2039/5254 ,  A61K2039/5256 ,  C07K14/005 ,  C12N15/86 ,  C12N2740/16222 ,  C12N2760/20243 ,  C12N2760/20251 ,  C12N2760/20252 ,  Y02A50/464

Abstract: Novel purification processes for obtaining vesicular stomatitis virus (VSV) of improved purity from mammalian cell culture are described herein. More particularly, in certain embodiments, a process is described for purifying VSV from cell culture fluid of a mammalian cell culture infected with VSV, the process comprising: clarifying the cell culture fluid by low-speed centrifugation and recovering the VSV in the supernatant; filtering the supernatant through a 0.2 to 0.45 μm filter and recovering the VSV in the filtered solution; loading the VSV filtered solution onto a anion exchange membrane adsorber equilibrated with a first pH buffered salt solution, eluting the VSV from the anion exchange membrane adsorber with a second pH buffered salt solution and recovering the eluted VSV fractions; purifying the recovered VSV by tangential flow filtration (TFF) using a TFF membrane having a molecular weight cutoff between 300 kDa and 1,000 kDa and recovering the VSV in the retentate, and filtering the VSV retentate through a 0.2 to 0.22 μm filter and recovering the VSV in the filtered solution.

Abstract translation: 本文描述了从哺乳动物细胞培养物获得改善纯度的水泡性口炎病毒（VSV）的新型纯化方法。 更具体地，在某些实施方案中，描述了用于从感染VSV的哺乳动物细胞培养物的细胞培养液中纯化VSV的方法，该方法包括：通过低速离心澄清细胞培养液并回收上清液中的VSV; 通过0.2至0.45 mum过滤器过滤上清液并回收过滤溶液中的VSV; 将VSV过滤的溶液加载到用第一pH缓冲盐溶液平衡的阴离子交换膜吸附器上，用第二pH缓冲盐溶液从阴离子交换膜吸附器洗脱VSV并回收洗脱的VSV级分; 通过切向流过滤（TFF）使用分子量截留值在300kDa至1,000kDa之间的TFF膜来纯化回收的VSV，并回收渗余物中的VSV，并通过0.2至0.22mum的过滤器过滤VSV滞留物并回收VSV 在过滤的溶液中。

公开(公告)号：US07875446B2

公开(公告)日：2011-01-25

申请号：US11788071

申请日：2007-04-19

Applicant: Yun Kang ,  Mark William Cutler ,  Amadou Affrey Ouattara ,  Kristen Elissa Syvertsen

Inventor： Yun Kang ,  Mark William Cutler ,  Amadou Affrey Ouattara ,  Kristen Elissa Syvertsen

IPC: C12N7/02 ,  C12N1/02 ,  C12N7/01 ,  A61K39/205 ,  A61K39/38 ,  A61K39/12 ,  A01N63/00 ,  A61K39/23

CPC classification number: C12N7/00 ,  A61K39/00 ,  A61K48/00 ,  A61K2039/5252 ,  A61K2039/5254 ,  A61K2039/5256 ,  C07K14/005 ,  C12N15/86 ,  C12N2740/16222 ,  C12N2760/20243 ,  C12N2760/20251 ,  C12N2760/20252 ,  Y02A50/464

Abstract: A process is described for purifying vesicular stomatitis virus (VSV) from cell culture fluid of a mammalian cell culture infected with VSV, the process comprising: clarifying the cell culture fluid by low-speed centrifugation and recovering the VSV in the supernatant; filtering the supernatant through a 0.2 to 0.45 μm filter and recovering the VSV in the filtered solution; loading the VSV filtered solution onto a anion exchange membrane adsorber equilibrated with a first pH buffered salt solution, eluting the VSV from the anion exchange membrane adsorber with a second pH buffered salt solution and recovering the eluted VSV fractions; purifying the recovered VSV by tangential flow filtration (TFF) using a TFF membrane having a molecular weight cutoff between 300 kDa and 1,000 kDa and recovering the VSV in the retentate, and filtering the VSV retentate through a 0.2 to 0.22 μm filter and recovering the VSV in the filtered solution.

Abstract translation: 描述了从感染VSV的哺乳动物细胞培养物的细胞培养液中纯化水泡性口炎病毒（VSV）的方法，该方法包括：通过低速离心澄清细胞培养液并回收上清液中的VSV; 通过0.2至0.45μm的过滤器过滤上清液并在过滤的溶液中回收VSV; 将VSV过滤的溶液加载到用第一pH缓冲盐溶液平衡的阴离子交换膜吸附器上，用第二pH缓冲盐溶液从阴离子交换膜吸附器洗脱VSV并回收洗脱的VSV级分; 通过切向流过滤（TFF），使用分子量截止值在300kDa至1,000kDa之间的TFF膜来纯化回收的VSV，并回收渗余物中的VSV，并通过0.2至0.22μm的过滤器过滤VSV滞留物并回收VSV 在过滤的溶液中。

公开(公告)号：US20090175906A1

公开(公告)日：2009-07-09

申请号：US12338367

申请日：2008-12-18

Applicant: Narender Kumar Kalyan ,  Irina Yurgelonis ,  Roger Michael Hendry ,  Mark William Cutler ,  Kristen Elissa Syvertsen

Inventor： Narender Kumar Kalyan ,  Irina Yurgelonis ,  Roger Michael Hendry ,  Mark William Cutler ,  Kristen Elissa Syvertsen

IPC: A61K39/12 ,  C12N7/01 ,  C12N7/08

CPC classification number: C07D313/04 ,  A61K39/00 ,  A61K2039/5254 ,  A61K2039/5256 ,  C07K14/005 ,  C12N7/00 ,  C12N2760/20222 ,  C12N2760/20262 ,  C12N2760/20264 ,  Y02A50/412 ,  Y02A50/464 ,  Y02A50/466 ,  Y02A50/467

Abstract: The present invention relates to methods for generating genetically modified and attenuated strains of vesicular stomatitis virus (VSV) for use in the preparation of immunogenic compositions. More particularly, the invention relates to the identification of particular genetic modifications of attenuated VSV that result in an increased yield of virus and an increase in stability of the attenuated strains for preparation of the immunogenic compositions. Methods for cell culture propagation and use in large scale production of VSV is also disclosed.

Abstract translation: 本发明涉及用于产生用于制备免疫原性组合物的水泡性口炎病毒（VSV）的遗传修饰和减毒菌株的方法。 更具体地，本发明涉及鉴定导致增加的病毒产量和增强减毒菌株制备免疫原性组合物的稳定性的减毒VSV的特定遗传修饰。 还公开了用于细胞培养传播和用于大规模生产VSV的方法。