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    Efficient purification of adenovirus
    1.
    发明授权
    Efficient purification of adenovirus 有权

    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.

    Efficient purification of adenovirus
    2.
    发明授权
    Efficient purification of adenovirus 有权

    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.

    Fluorescence detection
    3.
    发明授权
    Fluorescence detection 有权

    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.

    Utilizing intrinsic fluorescence to detect adenovirus
    4.
    发明授权
    Utilizing intrinsic fluorescence to detect adenovirus 有权
    利用内在荧光检测腺病毒

    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。