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。

公开(公告)号：US20030143609A1

公开(公告)日：2003-07-31

申请号：US10336702

申请日：2003-01-03

Applicant: GenVec, Inc

Inventor： Miguel E. Carrion ,  Imre Kovesdi

IPC: C12Q001/68 ,  C12Q001/70

CPC classification number: C12N15/86 ,  C12N15/1055 ,  C12N2710/10343 ,  C12Q1/6816 ,  C12Q1/6897 ,  C12Q2563/167

Abstract: The present invention provides a method of identifying a gene product. The method comprises providing a multiplicity of cells comprising a first gene product. Preferably, the first gene product is produced in the multiplicity of cells by expressing a first exogenous nucleic acid sequence encoding the first gene product. A library of second nucleic acid sequences encoding second gene products is then introduced into the multiplicity of cells. The second nucleic acid sequences are expressed in the multiplicity of cells to produce the second gene products such that the first gene product and at least one of the second gene products contact. The method further comprises causing a complex to form between the first gene product, an affinity molecule that binds the first gene product, and at least one of the second gene products, and subsequently retrieving the complex. At least one second gene product of the complex then is identified.

Abstract translation: 本发明提供鉴定基因产物的方法。 该方法包括提供包含第一基因产物的多个细胞。 优选地，通过表达编码第一基因产物的第一外源核酸序列，在多种细胞中产生第一基因产物。 然后将编码第二基因产物的第二核酸序列的文库导入多个细胞。 第二核酸序列在多重细胞中表达以产生第二基因产物，使得第一基因产物和第二基因产物中的至少一个接触。 所述方法还包括使第一基因产物，结合第一基因产物的亲和分子与至少一种第二基因产物形成复合物，随后检索该复合物。 然后鉴定该复合物的至少一个第二基因产物。

公开(公告)号：US20020045160A1

公开(公告)日：2002-04-18

申请号：US09888681

申请日：2001-06-25

Applicant: GenVec, Inc.

Inventor： Miguel E. Carrion ,  Marilyn C. Menger

IPC: C12Q001/70 ,  C12N013/00 ,  G01N033/558 ,  G01N033/559 ,  G01N033/561

CPC classification number: G01N21/6486 ,  C12N15/86 ,  C12N2710/10343 ,  C12N2710/10351

Abstract: The present invention provides methods of detecting and/or characterizing the viral vector particle content of a medium. A medium is provided and contacted with an excitation energy such that, if a viral vector particle is in the medium, an electron associated with the intrinsically fluorogenic portion of the viral vector particle will be raised to an excited energy state. The excited electron is permitted to emit radiation having an emission wavelength which is detected. The viral vector particle content of the medium then can be evaluated by comparing the detected emission wavelength with a standard signal. For example, the number of viral vector particles in a medium can be quantified by comparing the detected wavelength and its corresponding intensity to a standard signal. Similar methods for evaluating the adenoviral vector particle content of a medium and the intrinsically fluorogenic adenoviral structural protein content of a medium are provided.

Abstract translation: 本发明提供了检测和/或表征介质的病毒载体颗粒含量的方法。 提供介质并与激发能接触，使得如果病毒载体颗粒在介质中，与病毒载体颗粒的固有荧光部分相关的电子将被升高到激发能态。 被激发的电子被允许发射具有检测到的发射波长的辐射。 然后可以通过将检测到的发射波长与标准信号进行比较来评估介质的病毒载体颗粒含量。 例如，可以通过将检测到的波长及其对应的强度与标准信号进行比较来量化介质中病毒载体颗粒的数量。 提供了用于评估培养基的腺病毒载体颗粒含量和介质的固有荧光腺病毒结构蛋白含量的类似方法。