公开(公告)号：US20030053989A1

公开(公告)日：2003-03-20

申请号：US09952498

申请日：2001-09-14

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi

IPC: A61K048/00 ,  C12N015/861

CPC classification number: A61K38/1866 ,  A61K38/179 ,  A61K38/18 ,  A61K38/1891 ,  A61K38/195 ,  A61K48/00 ,  C12N15/86 ,  C12N2710/10343 ,  A61K2300/00

Abstract: The present invention provides a method of modulating neovascularization in an animal. The method comprises administering to the animal two or more nucleic acid sequences, each nucleic acid sequence encoding at least one angiogenesis-modulation factor that acts upon a different angiogenic process, such that the nucleic acid sequences are expressed to produce the angiogenesis-modulation factors to modulate neovascularization in the animal. Modulating neovascularization includes the induction of neovascularization or, in the alternative, the inhibition or reduction of neovascularization.

Abstract translation: 本发明提供一种调节动物体内新血管形成的方法。 该方法包括向动物施用两个或更多个核酸序列，每个核酸序列编码至少一种血管生成调节因子，其作用于不同的血管生成过程，使得所述核酸序列被表达以产生血管生成调节因子 调节动物的新血管形成。 调节新血管形成包括新血管形成的诱导，或者替代地，抑制或减少新生血管形成。

公开(公告)号：US20030040100A1

公开(公告)日：2003-02-27

申请号：US09911020

申请日：2001-07-23

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Imre Kovesdi

IPC: C12N007/02 ,  C12N005/02 ,  C07H021/04 ,  C12N005/00 ,  C12N007/01 ,  C12N007/00

CPC classification number: C12N15/86 ,  C12N7/00 ,  C12N2710/10321 ,  C12N2710/10343 ,  C12N2710/10352

Abstract: The invention provides a cell and a method of using the cell for the propagation of a replication-deficient adenoviral vector, wherein the cellular genome comprises a nucleic acid sequence whose expression produces a gene product that complements a replication-deficient adenoviral vector. The nucleic acid sequence is operatively linked to a chimeric expression control sequence comprising at least a functional portion of a CMV immediate early promoter/enhancer region and/or at least a functional portion of an adenoviral promoter, wherein the chimeric expression control sequence is upregulated by one or more viral proteins not produced by the nucleic acid sequence.

Abstract translation: 本发明提供了一种细胞和使用该细胞用于增殖复制缺陷型腺病毒载体的方法，其中所述细胞基因组包含其表达产生补充复制缺陷型腺病毒载体的基因产物的核酸序列。 核酸序列可操作地连接到包含CMV立即早期启动子/增强子区的至少一个功能部分和/或腺病毒启动子的至少功能部分的嵌合表达控制序列，其中嵌合表达控制序列被上调 不是由核酸序列产生的一种或多种病毒蛋白。

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：US20020019041A1

公开(公告)日：2002-02-14

申请号：US09870920

申请日：2001-05-31

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Stephen C. Ransom

IPC: A61K039/235 ,  C12N007/01

CPC classification number: C12N15/86 ,  A61K47/183 ,  A61K47/26 ,  A61K47/32 ,  A61K48/0091 ,  C12N2710/10343 ,  C12N2710/10351

Abstract: The present invention provides a method and a composition for preserving a virus. The virus is placed in a liquid carrier with a stabilizing agent selected from the group consisting of polysorbate 80, L-arginine, polyvinylpyrrolidone, trehalose, and combinations thereof. The liquid composition can be maintained at a temperature above 0null C. for a significant period of time while maintaining a satisfactory degree of viral activity.

Abstract translation: 本发明提供了用于保存病毒的方法和组合物。 将病毒置于具有选自聚山梨醇酯80，L-精氨酸，聚乙烯吡咯烷酮，海藻糖及其组合的稳定剂的液体载体中。 液体组合物可以在高于0℃的温度下保持相当长的一段时间，同时保持令人满意的病毒活性程度。

公开(公告)号：US20020004242A1

公开(公告)日：2002-01-10

申请号：US09905758

申请日：2001-07-13

Applicant: GenVec, Inc.

Inventor： Duncan L. McVey ,  Douglas E. Brough ,  Imre Kovesdi

IPC: C12N015/86 ,  C07H021/04

CPC classification number: C12N15/86 ,  C12N15/65 ,  C12N2710/10343 ,  C12N2800/30 ,  C12N2830/001 ,  C12N2830/15 ,  C12N2830/55

Abstract: The present invention provides a dual selection cassette (DSC) comprising first and second DNA segments having homology to a eukaryotic viral vector, positive and negative selection genes, each operably linked to their own promoter, and one or more unique restriction enzyme sites (URES) or sitey-directed homologous recombination sites. The present invention also provides a plasmid, pN/P, comprising an independent positive selection marker gene, an origin of replication, and a dual selection cassette. The dual selection cassette and pN/P plasmid can be used to produce eukaryotic gene transfer vectors without requiring temporally-linked double recombination events or the use of specialized bacterial strains that allow the replication of plasmids comprising defective origins of replication. This method usefully increases the ratio of desired to undesired plasmid and vector constructs. Additionally, this invention provides a method for the creation of eukaryotic viral vector libraries.

Abstract translation: 本发明提供了一种双重选择盒（DSC），其包含与真核病毒载体具有同源性的第一和第二DNA片段，每个与其本身的启动子可操作地连接的阳性和阴性选择基因，以及一个或多个独特的限制酶位点（URES） 或定点同源重组位点。 本发明还提供了包含独立的阳性选择标记基因，复制起点和双选择盒的质粒pN / P。 双选择盒和pN / P质粒可用于产生真核基因转移载体，而不需要时间连接的双重重组事件或使用允许复制包含有缺陷复制起点的质粒的专门细菌菌株。 该方法有效地增加了期望的和不需要的质粒和载体构建体的比例。 此外，本发明提供了一种用于产生真核病毒载体文库的方法。

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：US20030153065A1

公开(公告)日：2003-08-14

申请号：US10046517

申请日：2002-01-14

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Stephen C. Ransom

IPC: C12N007/01

CPC classification number: C12N7/00 ,  C12N2710/10351

Abstract: The invention provides a composition and a method for preserving a non-enveloped viral vector. The composition comprises (a) trehalose, (b) a divalent metal salt, a cationic polymer, or a combination thereof, (c) a multiplicity of non-enveloped viral vector particles, and (d) a liquid carrier. Non-enveloped virus particles are stable in the composition in a liquid form, at elevated temperatures, for a sustained period of time.

Abstract translation: 本发明提供了一种用于保存非包膜病毒载体的组合物和方法。 组合物包含（a）海藻糖，（b）二价金属盐，阳离子聚合物或其组合，（c）多个非包膜病毒载体颗粒，和（d）液体载体。 无包膜的病毒颗粒在液体形式的组合物中在高温下持续持续一段时间是稳定的。

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：US20030087438A1

公开(公告)日：2003-05-08

申请号：US10001097

申请日：2001-11-02

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Imre Kovesdi

IPC: A61K048/00 ,  C12N015/861 ,  C12N007/02 ,  C12N007/00

CPC classification number: C12N15/86 ,  A61K48/00 ,  C12N2710/10343

Abstract: The invention provides a composition comprising particles of an adenoviral vector comprising deficiencies in two or more gene functions required for viral replication, wherein at least one of the deficiencies is of a gene function of the E1 region of the adenoviral genome and (b) a carrier therefor, with relatively high ratios of (i) the number of particles of the adenoviral vectors to the number of particles of E1-revertant replication-deficient adenoviral vectors not comprising one or more of the deficiencies in gene functions of the E1 region of the adenoviral and (ii) the number of particles of the adenoviral vectors to the number of particles of replication-competent adenoviral vectors, as well as a method of preparing such a composition.

Abstract translation: 本发明提供包含腺病毒载体颗粒的组合物，其包含病毒复制所需的两个或多个基因功能的缺陷，其中至少一个缺陷是腺病毒基因组的E1区域的基因功能，和（b）载体 因此，具有相对高的比例（i）腺病毒载体的颗粒数与E1复发缺失型复制缺陷型腺病毒载体的颗粒数量不包含腺病毒E1区的基因功能的一个或多个缺陷 和（ii）腺病毒载体的颗粒数与复制能力的腺病毒载体的颗粒数目，以及制备这种组合物的方法。

公开(公告)号：US20030045498A1

公开(公告)日：2003-03-06

申请号：US10211701

申请日：2002-08-02

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Douglas E. Brough ,  Lisa Wei ,  Duncan L. McVey

IPC: A61K048/00 ,  C12N015/85

CPC classification number: C12N15/86 ,  A61K48/00 ,  A61K48/005 ,  A61K48/0075 ,  C07K14/71 ,  C07K14/811 ,  C12N2710/10343 ,  C12N2799/022

Abstract: The present invention is directed to a method of prophylactically or therapeutically treating an animal for at least one ocular-related disorder, e.g., ocular neovascularization or age-related macular degeneration. The method comprises contacting an ocular cell with an expression vector comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and the same or different nucleic acid sequence encoding a neurotrophic agent. The method also can comprise contacting an ocular cell with different expression vectors, each comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and/or a nucleic acid sequence encoding a neurotrophic agent. In addition, the present invention provides a viral vector comprising a nucleic acid sequence encoding pigment epithelium-derived factor (PEDF) or a therapeutic fragment thereof.

Abstract translation: 本发明涉及一种预防性或治疗性地治疗动物至少一种与眼部相关的疾病，例如眼睛新生血管形成或年龄相关性黄斑变性的方法。 该方法包括使眼细胞与包含编码血管生成抑制剂的核酸序列和编码神经营养剂的相同或不同核酸序列的表达载体接触。 该方法还可以包括使眼细胞与不同的表达载体接触，每种表达载体包含编码血管生成抑制剂的核酸序列和/或编码神经营养剂的核酸序列。 此外，本发明提供了包含编码色素上皮衍生因子（PEDF）的核酸序列或其治疗片段的病毒载体。