Method for producing recombinant adeno-associated virus vectors
    2.
    发明授权
    Method for producing recombinant adeno-associated virus vectors 失效
    重组腺相关病毒载体的制备方法

    公开(公告)号:US5681731A

    公开(公告)日:1997-10-28

    申请号:US459049

    申请日:1995-06-02

    摘要: Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of viral elements produced by conventional methods.

    摘要翻译: 描述了产生重组腺相关病毒(rAAV)载体的简化方法。 该方法涉及使用掺入爱泼斯坦巴尔核抗原(EBNA)基因,爱泼斯坦巴尔病毒(oriP)的潜在复制起点)和rAAV基因组的嵌合质粒。 嵌合质粒本身也是本发明的一部分。 这些质粒作为细胞中的多拷贝染色体外元件维持,如人293细胞。 携带这些EBV / AAV质粒的永久性细胞系在添加野生型腺相关病毒辅助功能后被诱导产生大量的rAAV。 以这种方式产生的载体能够将外源基因转导到其他人细胞系中,并显示通过常规方法产生的病毒元件的属性。

    Stabilization of sterilized surfaces for research and medical use
    6.
    发明授权
    Stabilization of sterilized surfaces for research and medical use 失效
    用于研究和医疗用途的灭菌表面的稳定化

    公开(公告)号:US5283034A

    公开(公告)日:1994-02-01

    申请号:US858125

    申请日:1992-03-26

    摘要: A method and composition which permits sterilization of surfaces coupled with biologically active moieties by ionizing radiation is described. The protecting composition contains a surface-stabilizing agent which adheres to the surface and has a molecular weight.gtoreq.5 kd, and an oxygen radical scavenger which is preferably a di- or polysaccharide or reduced form thereof. In the method of the invention, a surface which is coupled to a biologically active agent is protected with the invention composition, dried to a moisture content of less than 1%, and then sterilized by ionizing radiation under standard conditions. The sterilized surfaces of the invention are particularly useful in the production of medical devices intended for extracorporeal use, particularly in cell-separation techniques.

    摘要翻译: 描述了允许通过电离辐射与生物活性部分偶联的表面灭菌的方法和组合物。 保护性组合物含有粘附到表面并具有≥5kd分子量的表面稳定剂,以及氧自由基清除剂,其优选为二糖或多糖或还原形式。 在本发明的方法中,与本发明组合物一起保护与生物活性剂偶联的表面,干燥至含水量小于1%,然后在标准条件下通过电离辐射灭菌。 本发明的灭菌表面特别可用于生产用于体外使用的医疗装置,特别是在细胞分离技术中。

    Device and process for cell capture and recovery
    8.
    发明授权
    Device and process for cell capture and recovery 失效
    用于细胞捕获和恢复的装置和过程

    公开(公告)号:US06432653B1

    公开(公告)日:2002-08-13

    申请号:US08291335

    申请日:1994-08-17

    申请人: Thomas B. Okarma

    发明人: Thomas B. Okarma

    IPC分类号: A61K3512

    摘要: Devices, processes and compositions are provided for effective separation of cells from a mixture of cells, where depletion or positive selection may be employed to provide a cellular population of interest. Of particular utility is the separation of cells from peripheral blood mononuclear cells, where members of the lymphoid or myeloid lineages may be isolated and used for research, diagnosis or therapy. Also of interest are cellular separation from bone marrow, tumor suspensions or lymphoid tissue suspensions, where cells can be isolated and used for a variety of purposes. The separated cells may be homogeneous, free of exogenous biologicals, viable, capable of replication and exhibit their full complement of biological activities. Multiple phenotypes can be captured simultaneously. Captured cells can be specifically activated with cytokines and antigens to provide cells which are MHC restricted and have antigen-specific effector functions.

    摘要翻译: 提供了装置,方法和组合物用于从细胞混合物中有效分离细胞,其中可以使用消耗或阳性选择来提供感兴趣的细胞群体。 特别有用的是将细胞与外周血单核细胞分离,其中淋巴或骨髓谱系的成员可以被分离并用于研究,诊断或治疗。 还有兴趣的是与骨髓,肿瘤悬浮液或淋巴组织悬浮液的细胞分离,其中细胞可以分离并用于各种目的。 分离的细胞可以是均匀的,没有外源生物学,可行的,能够复制并表现出其完整的生物活性。 可以同时捕获多种表型。 捕获的细胞可以用细胞因子和抗原特异性活化,以提供MHC限制的细胞并具有抗原特异性效应子功能。

    Device and process for cell capture and recovery

    公开(公告)号:US6143508A

    公开(公告)日:2000-11-07

    申请号:US484559

    申请日:1995-06-07

    申请人: Thomas B. Okarma

    发明人: Thomas B. Okarma

    摘要: Devices, processes and compositions are provided for effective separation of cells from a mixture of cells, where depletion or positive selection may be employed to provide a cellular population of interest. Of particular utility is the separation of cells from peripheral blood mononuclear cells, where members of the lymphoid or myeloid lineages may be isolated and used for research, diagnosis or therapy. Also of interest are cellular separation from bone marrow, tumor suspensions or lymphoid tissue suspensions, where cells can be isolated and used for a variety of purposes. The separated cells may be homogeneous, free of exogenous biologicals, viable, capable of replication and exhibit their full complement of biological activities. Multiple phenotypes can be captured simultaneously. Captured cells can be specifically activated with cytokines and antigens to provide cells which are MHC restricted and have antigen-specific effector functions.

    Recombinant adeno-associated virus vectors
    10.
    发明授权
    Recombinant adeno-associated virus vectors 失效
    重组腺相关病毒载体

    公开(公告)号:US5780280A

    公开(公告)日:1998-07-14

    申请号:US459352

    申请日:1995-06-02

    摘要: Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of viral elements produced by conventional methods.

    摘要翻译: 描述了产生重组腺相关病毒(rAAV)载体的简化方法。 该方法涉及使用掺入爱泼斯坦巴尔核抗原(EBNA)基因,爱泼斯坦巴尔病毒(oriP)的潜在复制起点)和rAAV基因组的嵌合质粒。 嵌合质粒本身也是本发明的一部分。 这些质粒作为细胞中的多拷贝染色体外元件维持,如人293细胞。 携带这些EBV / AAV质粒的永久性细胞系在添加野生型腺相关病毒辅助功能后被诱导产生大量的rAAV。 以这种方式产生的载体能够将外源基因转导到其他人细胞系中,并显示通过常规方法产生的病毒元件的属性。