公开(公告)号：US06383795B1

公开(公告)日：2002-05-07

申请号：US09296962

申请日：1999-04-22

申请人： Miguel E. Carrión ,  Marilyn Menger ,  Imre Kovesdi

发明人： Miguel E. Carrión ,  Marilyn Menger ,  Imre Kovesdi

IPC分类号： C12N702

CPC分类号： 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

摘要： 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.

公开(公告)号：US06630299B2

公开(公告)日：2003-10-07

申请号：US09888681

申请日：2001-06-25

申请人： Miguel E. Carrión ,  Marilyn Menger

发明人： Miguel E. Carrión ,  Marilyn Menger

IPC分类号： C12Q100

CPC分类号： G01N21/6486 ,  C12N15/86 ,  C12N2710/10343 ,  C12N2710/10351

摘要： 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.

公开(公告)号：US06447995B1

公开(公告)日：2002-09-10

申请号：US09679439

申请日：2000-10-04

申请人： Miguel E. Carrión ,  Marilyn Menger

发明人： Miguel E. Carrión ,  Marilyn Menger

IPC分类号： C12Q100

CPC分类号： G01N21/6486 ,  C12N15/86 ,  C12N2710/10343 ,  C12N2710/10351

摘要： 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.

摘要翻译： 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。

公开(公告)号：US06861229B2

公开(公告)日：2005-03-01

申请号：US10336702

申请日：2003-01-03

申请人： Miguel E. Carrión ,  Imre Kovesdi

发明人： Miguel E. Carrión ,  Imre Kovesdi

IPC分类号： C12N15/10 ,  C12N15/861 ,  C12Q1/68 ,  G01N33/53 ,  C12N15/63 ,  C12N15/70 ,  C11N15/861

CPC分类号： C12N15/86 ,  C12N15/1055 ,  C12N2710/10343 ,  C12Q1/6816 ,  C12Q1/6897 ,  C12Q2563/167

摘要： 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.

摘要翻译： 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：US06586226B2

公开(公告)日：2003-07-01

申请号：US09997909

申请日：2001-11-30

申请人： Miguel E. Carrión ,  Marilyn Menger ,  Imre Kovesdi

发明人： Miguel E. Carrión ,  Marilyn Menger ,  Imre Kovesdi

IPC分类号： C12N702

CPC分类号： 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

摘要： 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.