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    Scalable production method for AAV
    2.
    发明授权
    Scalable production method for AAV 有权
    AAV可扩展生产方法

    公开(公告)号：US09198984B2

    公开(公告)日：2015-12-01

    申请号：US12226588

    申请日：2007-04-27

    申请人： Martin Lock ,  Luc H Vandenberghe ,  James M. Wilson

    发明人： Martin Lock ,  Luc H Vandenberghe ,  James M. Wilson

    IPC分类号： A61K38/00 ,  C12N7/00 ,  A61K48/00 ,  C07K14/005

    CPC分类号： C12N7/00 ,  A61K48/0091 ,  C07K14/005 ,  C12N2750/14151

    摘要： A method for producing AAV, without requiring cell lysis, is described. The method involves harvesting AAV from the supernatant. For AAV having capsids with a heparin binding site, the method involves modifying the AAV capsids and/or the culture conditions to ablate the binding between the AAV heparin binding site and the cells, thereby allowing the AAV to pass into the supernatant, i.e., media. Thus, the method of the invention provides supernatant containing high yields of AAV which have a higher degree of purity from cell membranes and intracellular materials, as compared to AAV produced using methods using a cell lysis step.

    摘要翻译： 描述了不需要细胞裂解的生产AAV的方法。 该方法包括从上清液中收获AAV。 对于具有肝素结合位点的衣壳的AAV，该方法包括修饰AAV衣壳和/或培养条件以消除AAV肝素结合位点与细胞之间的结合，从而允许AAV进入上清液，即培养基 。 因此，与使用细胞裂解步骤的方法产生的AAV相比，本发明的方法提供了含有高产率的AAV的上清液，其具有比细胞膜和细胞内物质更高的纯度。

    METHODS FOR MANUFACTURE A CAPACITOR WITH THREE-DIMENSIONAL HIGH SURFACE AREA ELECTRODES
    4.
    发明申请
    METHODS FOR MANUFACTURE A CAPACITOR WITH THREE-DIMENSIONAL HIGH SURFACE AREA ELECTRODES 审中-公开
    一种具有三维高表面电极的电容器的制造方法

    公开(公告)号：US20140233153A1

    公开(公告)日：2014-08-21

    申请号：US14033479

    申请日：2013-09-22

    申请人： LIANG CHAI ,  ALAN RAE ,  JAMES M. WILSON

    发明人： LIANG CHAI ,  ALAN RAE ,  JAMES M. WILSON

    IPC分类号： H01G4/008 ,  H01G4/12

    CPC分类号： H01G4/10 ,  H01G4/005 ,  H01G4/008 ,  H01G4/12 ,  H01G4/1209 ,  H01G4/30 ,  Y10T29/43 ,  Y10T29/435

    摘要： A capacitor, and methods of its manufacture, having improved capacitance efficiency which results from increasing the effective area of an electrode surface are disclosed. An improved “three-dimensional” capacitor may be constructed with electrode layers having three-dimensional aspects at the point of interface with a dielectric such that portions of the electrode extend into the dielectric layer. Advantageously, embodiments of a three-dimensional capacitor drastically reduce the space footprint that is required in a circuit to accommodate the capacitor, when compared to current capacitor designs. Increased capacitance density may be realized without using high k (high constant) dielectric materials, additional “electrode—dielectric—electrode” arrangements in an ever increasing stack, or serially stringing together multiple capacitors.

    摘要翻译： 公开了一种电容器及其制造方法，其具有由于增加电极表面的有效面积而提高的电容效率。 可以在与电介质的界面点处具有三维方面的电极层来构造改进的“三维”电容器，使得电极的一部分延伸到电介质层中。 有利的是，与当前的电容器设计相比，三维电容器的实施例大大减少了电路中所需的容纳电容器的空间足迹。 可以在不使用高k（高常数）电介质材料的情况下实现增加的电容密度，而在不断增加的叠层中使用附加的“电极 - 电介质电极”布置，或者串联连接多个电容器。

    Method for detecting adeno-associated virus
    6.
    发明授权
    Method for detecting adeno-associated virus 有权
    腺相关病毒检测方法

    公开(公告)号：US08524446B2

    公开(公告)日：2013-09-03

    申请号：US12962793

    申请日：2010-12-08

    申请人： Guangping Gao ,  James M Wilson ,  Mauricio R. Alvira

    发明人： Guangping Gao ,  James M Wilson ,  Mauricio R. Alvira

    IPC分类号： C12Q1/70 ,  C12Q1/68 ,  C07H21/04

    CPC分类号： C12N15/86 ,  A61K38/177 ,  A61K38/4846 ,  A61K48/00 ,  C07K14/005 ,  C12N7/00 ,  C12N2750/14121 ,  C12N2750/14122 ,  C12N2750/14143 ,  C12N2750/14151 ,  C12N2750/14152 ,  C12N2750/14162 ,  C12N2750/14171 ,  C12N2830/008 ,  C12N2830/48 ,  C12N2830/85 ,  C12N2830/90 ,  C12Q1/701 ,  C12Q2600/158 ,  C12Y304/21022

    摘要： A method for detecting and isolating AAV sequences in a sample of DNA obtained from tissue or cells is provided, which sample contains DNA and proviral AAV. The method involves subjecting the sample containing DNA to amplification via polymerase chain reaction (PCR) using a first set of primers which specifically amplify a first AAV region. The first AAV region is characterized by having at least 250 nucleotides of AAV capsid nucleic acid sequence, a variable sequence flanked by a sequence of at least 18 nucleotides at the 5′ end of the first AAV region and a sequence of at least 18 nucleotides at the 3′ end of the first AAV region. Each of the 5′ and 3′ at least 18 nucleotides is the same over at least 9 consecutive nucleotides relative to corresponding sequences in an alignment of at least two AAV serotypes. Each of the sets of primers consist of a 5′ primer and a 3′ primer. The method is further useful for identifying AAV sequences in the sample by the presence of amplified proviral AAV sequences.

    摘要翻译： 提供了从组织或细胞获得的DNA样品中检测和分离AAV序列的方法，该样品含有DNA和前病毒AAV。 该方法包括使用特异性扩增第一AAV区域的第一组引物通过聚合酶链反应（PCR）使含有DNA的样品进行扩增。 第一个AAV区的特征在于具有AAV衣壳核酸序列的至少250个核苷酸，在第一AAV区的5'末端侧接至少18个核苷酸的序列的可变序列和至少18个核苷酸的序列 第一个AAV区域的3'端。 相对于至少两种AAV血清型的比对中的相应序列，5'和3'至少18个核苷酸中的每一个在至少9个连续核苷酸上相同。 每组引物由5'引物和3'引物组成。 该方法进一步用于通过扩增的前病毒AAV序列的存在来鉴定样品中的AAV序列。