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    Apparatus for active biological sample preparation
    1.
    发明授权
    Apparatus for active biological sample preparation 失效

    公开(公告)号：US06824740B1

    公开(公告)日：2004-11-30

    申请号：US09561318

    申请日：2000-04-28

    申请人： Edward L. Sheldon, III ,  Thomas R. Jackson ,  Paul D. Swanson ,  Bradley S. Scott ,  Michael J. Heller

    发明人： Edward L. Sheldon, III ,  Thomas R. Jackson ,  Paul D. Swanson ,  Bradley S. Scott ,  Michael J. Heller

    IPC分类号： B01L1100

    CPC分类号： B01D57/02 ,  B01J19/0093 ,  B01L3/502753 ,  B01L2200/0631 ,  B01L2200/0668 ,  B01L2300/0636 ,  B01L2300/0645 ,  B01L2300/0816 ,  B01L2300/0877 ,  B01L2400/0421 ,  B01L2400/0424 ,  B01L2400/0644 ,  B03C5/026 ,  B03C5/028 ,  C12N13/00 ,  G01N27/44743 ,  H01L29/6656

    摘要： Systems and methods for the electronic sample preparation of biological materials utilize the differential charge-to-mass ratio and/or the differential affinity of sample constituents to separation materials for sample preparation. An integrated system is provided for performing some or all of the processes of: receipt of biological materials, cell selection, sample purification, sample concentration, buffer exchange, complexity reduction and/or diagnosis and analysis. In one embodiment, one or more sample chambers adapted to receive a buffer solution are formed adjacent to a spacer region which may include a trap or other affinity material, electrophoretic motion of the materials to be prepared being effected through operation of electrodes. In another aspect of this invention, a transporter or dipstick serves to collect and permit transport of materials, such as nucleic acids, most preferably DNA and/or RNA. In one embodiment, a membrane or trap is held in a frame which is adapted to mate with a channel formed in the spacer region. In another aspect of this invention, an electrophoretic system for biological sample preparation is operated in a manner so as to utilize the differential charge-to-mass ratio so as to control the migration of materials within the solution. In one aspect, bunching of selected materials is achieved by operation of two electrodes in a manner so as to reduce the spatial dispersion of those materials. In another aspect of this invention, a vertically disposed sample preparation unit includes an upper reservoir including and a collection chamber. A sample is preferably pre-prepared and densified, applied to the conductive polymer, electrophoresed so as to move nucleic acids into the conductive polymer and move undesired material away from the conductive polymer. Integrated systems are described in which cell separation, purification, complexity reduction and diagnosis may be performed together. In the preferred embodiment, cell separation and sample purification are performed in a first region, the steps of denaturation, complexity reduction and diagnosis being performed in a second region.

    Apparatus and methods for active biological sample preparation
    2.
    发明授权
    Apparatus and methods for active biological sample preparation 失效

    公开(公告)号：US06129828A

    公开(公告)日：2000-10-10

    申请号：US709358

    申请日：1996-09-06

    申请人： Edward L. Sheldon, III ,  Thomas R. Jackson ,  Paul D. Swanson ,  Bradley S. Scott ,  Michael J. Heller

    发明人： Edward L. Sheldon, III ,  Thomas R. Jackson ,  Paul D. Swanson ,  Bradley S. Scott ,  Michael J. Heller

    IPC分类号： B01J19/00 ,  B01L3/00 ,  B03C5/02 ,  C12N13/00 ,  G01N27/447 ,  B01D57/02 ,  B01D61/42 ,  C12N15/10 ,  G01N27/26

    CPC分类号： B01D57/02 ,  B01J19/0093 ,  B01L3/502753 ,  B03C5/026 ,  B03C5/028 ,  C12N13/00 ,  G01N27/44743 ,  H01L29/6656 ,  B01L2200/0631 ,  B01L2200/0668 ,  B01L2300/0636 ,  B01L2300/0645 ,  B01L2300/0816 ,  B01L2300/0877 ,  B01L2400/0421 ,  B01L2400/0424 ,  B01L2400/0644

    摘要： Systems and methods for the electronic sample preparation of biological materials utilize the differential charge-to-mass ratio and/or the differential affinity of sample constituents to separation materials for sample preparation. An integrated system is provided for performing some or all of the processes of: receipt of biological materials, cell selection, sample purification, sample concentration, buffer exchange, complexity reduction and/or diagnosis and analysis. In one embodiment, one or more sample chambers adapted to receive a buffer solution are formed adjacent to a spacer region which may include a trap or other affinity material, electrophoretic motion of the materials to be prepared being effected through operation of electrodes. In another aspect of this invention, a transporter or dipstick serves to collect and permit transport of materials, such as nucleic acids, most preferably DNA and/or RNA. In one embodiment, a membrane or trap is held in a frame which is adapted to mate with a channel formed in the spacer region. In another aspect of this invention, an electrophoretic system for biological sample preparation is operated in a manner so as to utilize the differential charge-to-mass ratio so as to control the migration of materials within the solution. In one aspect, bunching of selected materials is achieved by operation of two electrodes in a manner so as to reduce the spatial dispersion of those materials. In another aspect of this invention, a vertically disposed sample preparation unit includes an upper reservoir including and a collection chamber. A sample is preferably pre-prepared and densified, applied to the conductive polymer, electrophoresed so as to move nucleic acids into the conductive polymer and move undesired material away from the conductive polymer. Integrated systems are described in which cell separation, purification, complexity reduction and diagnosis may be performed together. In the preferred embodiment, cell separation and sample purification are performed in a first region, the steps of denaturation, complexity reduction and diagnosis being performed in a second region.

    Channel-less separation of bioparticles on a bioelectronic chip by dielectrophoresis
    7.
    发明授权
    Channel-less separation of bioparticles on a bioelectronic chip by dielectrophoresis 有权
    通过介电电泳在生物电子芯片上无生物颗粒的分离

    公开(公告)号：US06280590B1

    公开(公告)日：2001-08-28

    申请号：US09548522

    申请日：2000-04-13

    申请人： Jing Cheng ,  Edward L. Sheldon, III ,  Lei Wu ,  James P. O'Connell

    发明人： Jing Cheng ,  Edward L. Sheldon, III ,  Lei Wu ,  James P. O'Connell

    IPC分类号： G01N2726

    CPC分类号： B01L3/5027 ,  B01D57/02 ,  B01J19/0093 ,  B01J2219/00853 ,  B01J2219/00914 ,  B01L3/502753 ,  B01L3/502761 ,  B01L2200/0631 ,  B01L2200/0647 ,  B01L2200/0668 ,  B01L2200/10 ,  B01L2300/0636 ,  B01L2300/0645 ,  B01L2300/0816 ,  B01L2300/0877 ,  B01L2400/0415 ,  B01L2400/0421 ,  B01L2400/0424 ,  B01L2400/0496 ,  B01L2400/0644 ,  B03C5/026 ,  B03C5/028 ,  C12N13/00 ,  G01N27/44743 ,  H01L29/6656

    摘要： The present invention comprises devices and methods for performing channel-less separation of cell particles by dielectrophoresis, DC high voltage-pulsed electronic lysis of separated cells, separation of desired components from crude mixtures such as cell lysates, and/or enzymatic reaction of such lysates, all of which can be conducted on a single bioelectronic chip. A preferred embodiment of the present invention comprises a cartridge (10) including a microfabricated silicon chip (12) on a printed circuit board (14) and a flow cell (16) mounted to the chip (12) to form a flow chamber. The cartridge (10) also includes output pins (22) for electronically connecting the cartridge (10) to an electronic controller. The chip (12) includes a plurality of circular microelectrodes (24) which are preferably coated with a protective permeation layer which prevents direct contact between any electrode and a sample introduced into the flow chamber. The permeation layer also helps to reduce cell adhesion at field minima, and enables immobilization of specific antibodies for specific cell capture. Specific cells from various cell mixtures were separated, lysed, and enzymatically digested on the chip.

    摘要翻译： 本发明包括用于通过介电电泳，DC分离细胞的DC高压脉冲电离裂解，细胞裂解物等粗制混合物分离所需组分和/或这些裂解物的酶反应来进行细胞颗粒的无通道分离的装置和方法 所有这些都可以在单个生物电子芯片上进行。 本发明的优选实施例包括在印刷电路板（14）上包括微细硅芯片（12）的盒（10）和安装到芯片（12）上以形成流动室的流动池（16）。 盒（10）还包括用于将盒（10）电连接到电子控制器的输出销（22）。 芯片（12）包括多个圆形微电极（24），其优选地涂覆有保护性渗透层，其防止任何电极和引入流动室中的样品之间的直接接触。 渗透层还有助于减少最小场的细胞粘附，并且能够固定特异性抗体用于特异性细胞捕获。 分离来自各种细胞混合物的特异性细胞，裂解，并在芯片上酶消化。

    Process for labeling nucleic acids and hybridization probes
    9.
    发明授权
    Process for labeling nucleic acids and hybridization probes 失效
    标记核酸和杂交探针的方法

    公开(公告)号：US4617261A

    公开(公告)日：1986-10-14

    申请号：US791332

    申请日：1985-10-25

    申请人： Edward L. Sheldon, III ,  Corey H. Levenson ,  Kary B. Mullis ,  Henry Rapoport ,  Robert M. Watson

    发明人： Edward L. Sheldon, III ,  Corey H. Levenson ,  Kary B. Mullis ,  Henry Rapoport ,  Robert M. Watson

    IPC分类号： C07D493/04 ,  C07D513/04 ,  C07D519/00 ,  C07H21/00 ,  C12Q1/68 ,  G01N33/48

    CPC分类号： C07D493/04 ,  C07D513/04 ,  C07H21/00 ,  C12Q1/68 ,  Y10T436/143333

    摘要： Nucleic acids may be labeled by intercalating the alkylating intercalation moiety of a labeling reagent into a partially double-stranded nucleic acid to form a complex and activating the complex to cause covalent bonding between the reagent and the nucleic acid. Preferably, the labeled nucleic acid is a hybridization probe for detecting nucleic acid sequences capable of hybridizing with a hybridizing region of the nucleic acid. Also preferably the label moiety is non-radioactive. The labeling reagent is of the formula:[A--[B--Lwhere A is an alkylating intercalation moiety, B is a divalent organic moiety of the formula: ##STR1## where Y is O, NH or N--CHO, x is a number from 1 to 4, y is a number from 2 to 4, and L is a monovalent label moiety, wherein B is exclusive of any portion of the intercalation and label moieties.Preferably A is a 4-methylene-substituted psoralen moiety, and most preferably A is a 4'-methylene-substituted-4,5',8-trimethylpsoralen moeity and L is biotin.

    摘要翻译： 可以通过将标记试剂的烷基化插层部分嵌入部分双链核酸中以形成复合物并激活复合物以引起试剂和核酸之间的共价键来标记核酸。 优选地，标记的核酸是用于检测能够与核酸的杂交区域杂交的核酸序列的杂交探针。 还优选地，标记部分是非放射性的。 标记试剂具有下式：[A- [BL，其中A是烷基化插层部分，B是下式的二价有机部分：其中Y是O，NH或N-CHO，x是从 1至4，y为2至4的数，L为单价标记部分，其中B不包括插层和标记部分的任何部分。 优选A是4-亚甲基取代的补骨脂素部分，最优选A是4'-亚甲基取代的-4,5'，8-三甲基肉桂酸酯，L是生物素。