公开(公告)号：US20160102320A1

公开(公告)日：2016-04-14

申请号：US14879318

申请日：2015-10-09

Applicant: GENVEC, INC.

Inventor： Douglas E. Brough ,  Damodar R. Ettyreddy

IPC: C12N15/86 ,  A61K48/00 ,  A61K38/16

CPC classification number: C12N15/86 ,  A61K38/16 ,  A61K48/00 ,  A61K48/0058 ,  A61K48/0075 ,  C12N2710/10043 ,  C12N2710/10343 ,  C12N2830/008

Abstract: The invention is directed to a replication-deficient adenoviral vector comprising a nucleic acid sequence encoding a human atonal homolog-1 (Hath1) protein operably linked to a human glial fibrillary acidic protein (GFAP) promoter. The invention also is directed to a composition and method utilizing the adenoviral vector to generate sensory cells in the inner ear of a human.

Abstract translation: 本发明涉及复制缺陷型腺病毒载体，其包含编码与人胶质纤维酸性蛋白（GFAP）启动子可操作地连接的人类非同源同源物-1（Hath1）蛋白的核酸序列。 本发明还涉及利用腺病毒载体在人的内耳中产生感觉细胞的组合物和方法。

公开(公告)号：US20150329834A1

公开(公告)日：2015-11-19

申请号：US14817910

申请日：2015-08-04

Applicant: GenVec, Inc.

Inventor： Jason Gall ,  Douglas Brough ,  Christoph Kahl ,  Duncan McVey

IPC: C12N7/00

CPC classification number: C07K14/005 ,  C12N7/00 ,  C12N15/86 ,  C12N2710/10322 ,  C12N2710/10332 ,  C12N2710/10343 ,  C12N2710/10352

Abstract: The invention provides methods for propagating a monkey adenovirus in a cell, including a human cell, comprising one or more gene products isolated from a human adenovirus. Also provided are methods for propagating wherein the monkey adenovirus comprises a nucleic acid sequence encoding a human adenovirus gene product. The invention further provides a monkey adenovirus, including a replication-deficient monkey adenovirus, obtained by such propagation methods.

Abstract translation: 本发明提供了在细胞中繁殖猴腺病毒的方法，包括人细胞，其包含从人腺病毒分离的一种或多种基因产物。 还提供了用于繁殖的方法，其中猴腺病毒包含编码人腺病毒基因产物的核酸序列。 本发明还提供了通过这种繁殖方法获得的猴腺病毒，其包括复制缺陷型猴腺病毒。

公开(公告)号：US20030203480A1

公开(公告)日：2003-10-30

申请号：US10427759

申请日：2003-04-30

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Duncan L. McVey

IPC: C12N015/861 ,  C12N007/00

CPC classification number: C12N15/86 ,  C12N2710/10043 ,  C12N2795/10043 ,  C12N2810/80

Abstract: The inventive method of producing a eukaryotic viral vector comprises contacting a eukaryotic cell, which comprises a unique enzyme that nicks or cleaves a DNA molecule, with a recombinant phage vector, or contacting a eukaryotic cell, which does not comprise a unique enzyme that nicks or cleaves a DNA molecule, simultaneously or sequentially, in either order, with (i) a unique enzyme that nicks or cleaves a DNA molecule, and (ii) a recombinant phage vector. The recombinant phage vector comprises the DNA molecule comprising (a) a eukaryotic viral vector genome comprising a coding sequence, (b) a phage packaging site that is not contained within the eukaryotic viral vector genome, and (c) a promoter that is operably linked to the coding sequence. Alternatively, the DNA molecule is not present within the recombinant phage vector. The eukaryotic cell is contacted with the first DNA molecule and a recombinant phage vector. The first DNA molecule comprises a replication deficient eukaryotic viral vector genome comprising at least one adenoviral inverted terminal repeat and a packaging signal. The recombinant phage vector comprises a second DNA molecule and a phage packaging site, wherein the second DNA molecule complements in trans the replication deficient eukaryotic viral vector genome. The DNA molecule(s) enter the eukaryotic cell, and the unique enzyme nicks or cleaves the DNA molecule in the eukaryotic cell in at least one region not contained within the eukaryotic viral vector genome, thereby inducing the production of and ultimately producing a eukaryotic viral vector.

Abstract translation: 产生真核病毒载体的本发明方法包括将包含切割或切割DNA分子的独特酶与重组噬菌体载体接触的真核细胞或接触真核细胞，所述真核细胞不包含缺陷或 以（i）切割或切割DNA分子的独特的酶，和（ii）重组噬菌体载体以任一顺序同时或顺序地切割DNA分子。 重组噬菌体载体包含DNA分子，其包含（a）真核病毒载体基因组，其包含编码序列，（b）不包含在真核病毒载体基因组内的噬菌体包装位点，和（c）可操作地连接的启动子 到编码序列。 或者，DNA分子不存在于重组噬菌体载体内。 将真核细胞与第一DNA分子和重组噬菌体载体接触。 第一DNA分子包含复制缺陷型真核病毒载体基因组，其包含至少一个腺病毒反向末端重复和包装信号。 重组噬菌体载体包含第二DNA分子和噬菌体包装位点，其中第二DNA分子与复制缺陷型真核病毒载体基因组相反。 DNA分子进入真核细胞，独特的酶在至少一个不包含在真核病毒载体基因组内的区域中切割或切割真核细胞中的DNA分子，由此诱导产生并最终产生真核病毒 向量。

公开(公告)号：US20030086913A1

公开(公告)日：2003-05-08

申请号：US10003624

申请日：2001-11-02

Applicant: GenVec, Inc.

Inventor： Bryan Butman ,  Stephen Morris

IPC: A61K048/00 ,  C12N015/86

CPC classification number: A61K48/0091 ,  C12N2710/10343 ,  C12N2710/10351

Abstract: The invention provides a composition comprising a gene transfer vector which comprises a nucleic acid sequence encoding a protein and a carrier therefore. The composition is characterized by a relatively high ratio of the gene transfer vector to the protein in the composition.

Abstract translation: 本发明提供包含基因转移载体的组合物，其包含编码蛋白质和载体的核酸序列。 组合物的特征在于组合物中基因转移载体与蛋白质的比例相对较高。

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：US11034975B2

公开(公告)日：2021-06-15

申请号：US16282924

申请日：2019-02-22

Applicant: GenVec, Inc.

Inventor： Jason G. D. Gall ,  Duncan McVey ,  Douglas E. Brough

IPC: C12N15/86 ,  C07K14/005 ,  A61K39/155 ,  C12N7/00 ,  A61K48/00

Abstract: The invention provides an adenovirus or adenoviral vector characterized by comprising one or more particular nucleic acid sequences or one or more particular amino acid sequences, or portions thereof, pertaining to, for example, an adenoviral pIX protein, DNA polymerase protein, penton protein, hexone protein, and/or fiber protein.