公开(公告)号：US20020034735A1

公开(公告)日：2002-03-21

申请号：US09997909

申请日：2001-11-30

Applicant: GenVec, Inc.

Inventor： Miguel E. Carrion ,  Marilyn Menger ,  Imre Kovesdi

IPC: C12Q001/70 ,  C12N007/02 ,  C12N007/00 ,  C12N007/01

CPC classification number: C07K14/005 ,  B01D15/345 ,  B01D15/363 ,  B01J41/20 ,  C12N7/00 ,  C12N2710/10322 ,  C12N2710/10351 ,  C12Q1/06 ,  G01N30/06 ,  G01N30/96 ,  G01N2030/625 ,  G01N2030/8813 ,  G01N2333/075

Abstract: A method of enriching a solution of an adenovirus comprising applying a mixed solution comprising an adenovirus and at least one undesired type of biomolecule to an anion exchange chromatography resin containing a binding moiety selected from the group consisting of dimethylaminopropyl, dimethylaminobutyl, dimethylaminoisobutyl, and dimethylaminopentyl and eluting the adenovirus from the chromatography resin. Also provided is a method of purifying an adenovirus from adenovirus-infected cells comprising lysing such cells, applying the lysate to a single chromatography resin, eluting the adenovirus from the chromatography resin, and collecting a fraction containing adenovirus that is substantially as pure as triple CsCl density gradient-purified adenovirus. The present method further provides a method of accurately quantifying the number of adenoviral particles in a solution of adenovirus comprising applying to and eluting from an anion exchange chromatography resin a sample solution of adenovirus, comparing the absorbance of the sample solution of adenovirus and the absorbance of a standard solution of adenovirus, and quantifying the number of adenoviral particles in the sample solution.

Abstract translation: 一种富含腺病毒溶液的方法，包括将包含腺病毒和至少一种不期望类型的生物分子的混合溶液施加到含有选自二甲基氨基丙基，二甲基氨基丁基，二甲基氨基异丁基和二甲基氨基戊基的结合部分的阴离子交换色谱树脂和 从色谱树脂洗脱腺病毒。 还提供了一种从腺病毒感染的细胞中纯化腺病毒的方法，包括裂解这些细胞，将裂解物应用于单层色谱树脂，从色谱树脂洗脱腺病毒，并收集含有基本上与三重CsCl纯度相同的腺病毒的级分 密度梯度纯化腺病毒。 本方法还提供了一种准确地量化腺病毒溶液中腺病毒颗粒数量的方法，包括从阴离子交换层析树脂中施用和洗脱腺病毒样品溶液，并比较腺病毒样品溶液的吸光度和吸光度 腺病毒的标准溶液，并定量样品溶液中腺病毒颗粒的数量。

公开(公告)号：US20030203469A1

公开(公告)日：2003-10-30

申请号：US10429303

申请日：2003-05-05

Applicant: GenVec, Inc.

Inventor： Miguel E. Carrion ,  Marilyn Menger ,  Imre Kovesdi

IPC: C12N007/02

CPC classification number: C07K14/005 ,  B01D15/345 ,  B01D15/363 ,  B01J41/20 ,  C12N7/00 ,  C12N2710/10322 ,  C12N2710/10351 ,  C12Q1/06 ,  G01N30/06 ,  G01N30/96 ,  G01N2030/625 ,  G01N2030/8813 ,  G01N2333/075

Abstract: A method of enriching a solution for an adenovirus comprising contacting a solution containing an adenovirus with an anion exchange chromatography resin comprising an acrylate or sulphonamide linker such that the adenovirus binds to the chromatography resin and eluting the adenovirus from the resin with an eluant to obtain an enriched solution of adenovirus.

Abstract translation: 一种富含腺病毒溶液的方法，包括使含有腺病毒的溶液与包含丙烯酸酯或磺酰胺接头的阴离子交换层析树脂接触，使得腺病毒与色谱树脂结合并用洗脱剂从树脂中洗脱出腺病毒，得到 富集腺病毒溶液。