公开(公告)号：WO2008148010A8

公开(公告)日：2010-02-18

申请号：PCT/US2008064726

申请日：2008-05-23

申请人： VGX PHARMACEUTICALS INC ,  DRAGHIA-AKLI RUXANDRA ,  HEBEL HENRY ,  CAI YING

发明人： DRAGHIA-AKLI RUXANDRA ,  HEBEL HENRY ,  CAI YING

IPC分类号： G02B6/293

CPC分类号： C12N1/06 ,  C12N15/1017

摘要： Large scale processes for producing high purity samples of biologically active molecules of interest from bacterial cells are disclosed. The methods comprise the steps of producing a lysate solution by contacting a cell suspension of said plurality of cells with lysis solution; neutralizing said lysate solution with a neutralizing solution to produce a dispersion that comprises neutralized lysate solution and debris; filtering the dispersion through at least one filter; performing ion exchange separation on said neutralized lysate solution to produce an ion exchange eluate; and performing hydrophobic interaction separation on said ion exchange eluate to produce a hydrophobic interaction solution. Further, provided are compositions comprising large scale amounts of plasmid DNA produced by the disclosed large scale processes.

摘要翻译： 公开了从细菌细胞产生感兴趣的生物活性分子的高纯度样品的大规模方法。 所述方法包括通过使所述多个细胞的细胞悬浮液与裂解液接触来产生裂解物溶液的步骤; 用中和溶液中和所述裂解物溶液以产生包含中和的裂解液和碎屑的分散体; 通过至少一个过滤器过滤色散; 在所述中和的裂解物溶液上进行离子交换分离以产生离子交换洗脱液; 并在所述离子交换洗脱液上进行疏水相互作用分离以产生疏水相互作用溶液。 此外，提供了包含通过所公开的大规模方法生产的大量量的质粒DNA的组合物。

公开(公告)号：WO2008148010A1

公开(公告)日：2008-12-04

申请号：PCT/US2008064726

申请日：2008-05-23

申请人： VGX PHARMACEUTICALS INC ,  DRAGHIA-AKLI RUXANDRA ,  HEBEL HENRY ,  CAI YING

发明人： DRAGHIA-AKLI RUXANDRA ,  HEBEL HENRY ,  CAI YING

IPC分类号： G02B6/293

CPC分类号： C12N1/06 ,  C12N15/1017

摘要： Large scale processes for producing high purity samples of biologically active molecules of interest from bacterial cells are disclosed. The methods comprise the steps of producing a lysate solution by contacting a cell suspension of said plurality of cells with lysis solution; neutralizing said lysate solution with a neutralizing solution to produce a dispersion that comprises neutralized lysate solution and debris; filtering the dispersion through at least one filter; performing ion exchange separation on said neutralized lysate solution to produce an ion exchange eluate; and performing hydrophobic interaction separation on said ion exchange eluate to produce a hydrophobic interaction solution. Further, provided are compositions comprising large scale amounts of plasmid DNA produced by the disclosed large scale processes.

摘要翻译： 公开了从细菌细胞产生感兴趣的生物活性分子的高纯度样品的大规模方法。 所述方法包括通过使所述多个细胞的细胞悬浮液与裂解液接触来产生裂解物溶液的步骤; 用中和溶液中和所述裂解物溶液以产生包含中和的裂解液和碎屑的分散体; 通过至少一个过滤器过滤色散; 在所述中和的裂解物溶液上进行离子交换分离以产生离子交换洗脱液; 并在所述离子交换洗脱液上进行疏水相互作用分离以产生疏水相互作用溶液。 此外，提供了包含通过所公开的大规模方法生产的大量量的质粒DNA的组合物。

公开(公告)号：WO2004108260A3

公开(公告)日：2005-05-12

申请号：PCT/US2004016806

申请日：2004-05-27

申请人： ADVISYS INC ,  HEBEL HENRY ,  RAMAKRISHNAN SRIRAM ,  GONZALEZ HUGO ,  DARNELL JEFF

发明人： HEBEL HENRY ,  RAMAKRISHNAN SRIRAM ,  GONZALEZ HUGO ,  DARNELL JEFF

IPC分类号： B01F13/02 ,  C12N1/06 ,  C12N15/10 ,  B01F3/04

CPC分类号： C12N15/1003 ,  B01F13/0255 ,  B03D2203/003 ,  C12M47/06 ,  C12M47/10 ,  C12N1/06 ,  C12N1/066 ,  C12N15/101

摘要： An apparatus and a method for isolating a biologic product, such as plasmid DNA, from cells. The method involves lysing cells in a controlled manner separate insoluble components from a fluid lysate containing cellular components of interest, followed by membrane chromatographic techniques to purify the cellular components of interest. The process utilizes a unique lysis apparatus, ion exchange and, optionally, hydrophobic interaction chromatography membranes in cartridge form, and ultrafiltration. The process can be applied to any biologic product extracted from a cellular source. The process uses a lysis apparatus, including a high shear, low residence-time mixer for advantageously mixing a cell suspension with a lysis solution, a hold time that denatures impurities, and an air-sparging bubble mixer that gently yet thoroughly mixes lysed cells with a neutralization/precipiation buffer and floats compacted precipitated cellular material.

摘要翻译： 从细胞中分离生物制品如质粒DNA的装置和方法。 该方法包括以受控的方式裂解细胞，从含有感兴趣的细胞成分的流体裂解物中分离出不溶性组分，然后进行膜色谱技术纯化感兴趣的细胞组分。 该方法利用独特的裂解装置，离子交换以及任选的以胶囊形式的疏水相互作用层析膜和超滤。 该方法可以应用于从细胞来源提取的任何生物制品。 该方法使用裂解装置，包括高剪切，低停留时间混合器，有利地将细胞悬浮液与裂解溶液混合，使杂质变性的保持时间和将裂解细胞轻轻地充分混合的空气鼓泡混合器 中和/沉淀缓冲液和漂浮压实的沉淀的细胞材料。

公开(公告)号：WO2008148010A9

公开(公告)日：2008-12-04

申请号：PCT/US2008/064726

申请日：2008-05-23

申请人： VGX PHARMACEUTICALS, INC. ,  DRAGHIA-AKLI, Ruxandra ,  HEBEL, Henry ,  CAI, Ying

发明人： DRAGHIA-AKLI, Ruxandra ,  HEBEL, Henry ,  CAI, Ying

IPC分类号： C12N15/10

摘要： Large scale processes for producing high purity samples of biologically active molecules of interest from bacterial cells are disclosed. The methods comprise the steps of producing a lysate solution by contacting a cell suspension of said plurality of cells with lysis solution; neutralizing said lysate solution with a neutralizing solution to produce a dispersion that comprises neutralized lysate solution and debris; filtering the dispersion through at least one filter; performing ion exchange separation on said neutralized lysate solution to produce an ion exchange eluate; and performing hydrophobic interaction separation on said ion exchange eluate to produce a hydrophobic interaction solution. Further, provided are compositions comprising large scale amounts of plasmid DNA produced by the disclosed large scale processes.

公开(公告)号：WO2004108260A2

公开(公告)日：2004-12-16

申请号：PCT/US2004/016806

申请日：2004-05-27

申请人： ADVISYS, INC. ,  HEBEL, Henry ,  RAMAKRISHNAN, Sriram ,  GONZALEZ, Hugo ,  DARNELL, Jeff

发明人： HEBEL, Henry ,  RAMAKRISHNAN, Sriram ,  GONZALEZ, Hugo ,  DARNELL, Jeff

IPC分类号： B01F3/04

CPC分类号： C12N15/1003 ,  B01F13/0255 ,  B03D2203/003 ,  C12M47/06 ,  C12M47/10 ,  C12N1/06 ,  C12N1/066 ,  C12N15/101

摘要： An apparatus and a method for isolating a biologic product, such as plasmid DNA, from cells. The method involves lysing cells in a controlled manner separate insoluble components from a fluid lysate containing cellular components of interest, followed by membrane chromatographic techniques to purify the cellular components of interest. The process utilizes a unique lysis apparatus, ion exchange and, optionally, hydrophobic interaction chromatography membranes in cartridge form, and ultrafiltration. The process can be applied to any biologic product extracted from a cellular source. The process uses a lysis apparatus, including a high shear, low residence-time mixer for advantageously mixing a cell suspension with a lysis solution, a hold time that denatures impurities, and an air-sparging bubble mixer that gently yet thoroughly mixes lysed cells with a neutralization/precipiation buffer and floats compacted precipitated cellular material.

摘要翻译： 从细胞中分离生物制品如质粒DNA的装置和方法。 该方法包括以受控的方式裂解细胞，从含有感兴趣的细胞成分的流体裂解物中分离出不溶性组分，然后进行膜色谱技术纯化感兴趣的细胞组分。 该方法利用独特的裂解装置，离子交换以及任选的以胶囊形式的疏水相互作用层析膜和超滤。 该方法可以应用于从细胞来源提取的任何生物制品。 该方法使用裂解装置，包括高剪切，低停留时间混合器，有利地将细胞悬浮液与裂解溶液混合，使杂质变性的保持时间和将裂解细胞轻轻地充分混合的空气鼓泡混合器 中和/沉淀缓冲液和漂浮压实的沉淀的细胞材料。