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    Capacitive morpholino diagnostics for analysis of nucleic acids
    1.
    发明授权
    Capacitive morpholino diagnostics for analysis of nucleic acids 有权
    用于分析核酸的电容性吗啉诊断

    公开(公告)号：US08273578B2

    公开(公告)日：2012-09-25

    申请号：US13423228

    申请日：2012-03-18

    申请人： Rastislav Levicky ,  Napoleon Tercero ,  Kang Wong ,  Ping Gong ,  Kenneth Shepard

    发明人： Rastislav Levicky ,  Napoleon Tercero ,  Kang Wong ,  Ping Gong ,  Kenneth Shepard

    IPC分类号： G01N33/50

    CPC分类号： C12Q1/6825 ,  Y10T436/143333 ,  C12Q2525/113

    摘要： Use of Morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of Morpholino probes on gold supports are fabricated with methods similar to those employed with DNA, and are used to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for Morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a near-linear relationship, and demonstrates sensitivities (limits of quantification) in the pg mm−2 range.

    摘要翻译： 使用Morpholinos，一类不带电荷的DNA类似物，用于表面杂交应用。 使用类似于DNA使用的方法制备金支持物上的Morpholino探针的单层，并且用于与靶链有效地和序列特异性地杂交。 用电化学方法分析杂交诱导的界面电荷组织变化，并比较Morpholino和DNA探针单层。 通过与数值泊松 - 玻尔兹曼计算进行比较，可以确定和解释连接表面杂交态与界面电容的分子机制。 取决于由施加电位控制的移动离子的表面种群，对于杂交，阳性以及负电容反应（对比反转）是可能的。 表面电容与目标覆盖（目标/面积）的定量比较显示近似线性关系，并显示pg mm-2范围内的灵敏度（定量限度）。

    CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS
    2.
    发明申请
    CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS 有权
    用于核酸分析的电容性多元醇诊断

    公开(公告)号：US20100187133A1

    公开(公告)日：2010-07-29

    申请号：US12362071

    申请日：2009-01-29

    申请人： Rastislav Levicky ,  Napoleon Tercero ,  Kang Wang ,  Ping Gong ,  Kenneth Shepard

    发明人： Rastislav Levicky ,  Napoleon Tercero ,  Kang Wang ,  Ping Gong ,  Kenneth Shepard

    IPC分类号： G01N27/26

    CPC分类号： C12Q1/6825 ,  Y10T436/143333 ,  C12Q2525/113

    摘要： Surface hybridization, a reaction in which nucleic acid molecules in solution react with nucleic acid partners immobilized on a surface, is widely practiced in life science research. In these applications the immobilized partner, or “probe”, is typically single-stranded DNA. Because DNA is strongly charged, high salt conditions are required to enable binding between analyte nucleic acids (“targets”) in solution and the DNA probes. High salt, however, compromises prospects for label-free monitoring or control of the hybridization reaction through surface electric fields, as well as stabilizes secondary structure in target species that can interfere with probe-target recognition. In this work, initial steps toward addressing these challenges are taken by introducing Morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of Morpholino probes on gold supports can be fabricated with methods similar to those employed with DNA, and are shown to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for Morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Interestingly, positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a near-linear relationship, and demonstrates sensitivities (limits of quantification) in the pg mm−2 range.

    摘要翻译： 表面杂交，其中溶液中的核酸分子与固定在表面上的核酸配偶体反应的反应在生命科学研究中广泛应用。 在这些应用中，固定化的配偶体或“探针”通常是单链DNA。 由于DNA是强电荷的，因此需要高盐条件才能使溶液中的分析物核酸（“靶”）与DNA探针结合。 然而，高盐不利于通过表面电场进行无标记监测或控制杂交反应的前景，以及稳定可能干扰探针 - 目标识别的靶物种中的二级结构。 在这项工作中，通过引入Morpholinos（一类不带电荷的DNA类似物）进行表面杂交应用来采取解决这些挑战的最初步骤。 可以用与DNA一起使用的方法类似的方法制备金载体上的Morpholino探针的单层，并且显示与靶链有效地并且序列特异性地杂交。 用电化学方法分析杂交诱导的界面电荷组织变化，并比较Morpholino和DNA探针单层。 通过与数值泊松 - 玻尔兹曼计算进行比较，可以确定和解释连接表面杂交态与界面电容的分子机制。 有趣的是，取决于由施加电位控制的移动离子的表面种群，杂交的阳性和负电容反应（对比反转）是可能的。 表面电容与目标覆盖（目标/面积）的定量比较显示近似线性关系，并显示pg mm-2范围内的灵敏度（定量限度）。

    SOLID PHASE METHODS FOR THERMODYNAMIC AND KINETIC QUANTIFICATION OF INTERACTIONS BETWEEN NUCLEIC ACIDS AND SMALL MOLECULES
    5.
    发明申请
    SOLID PHASE METHODS FOR THERMODYNAMIC AND KINETIC QUANTIFICATION OF INTERACTIONS BETWEEN NUCLEIC ACIDS AND SMALL MOLECULES 审中-公开
    固体相对于核酸和小分子间相互作用的动态和动力学定量方法

    公开(公告)号：US20120295805A1

    公开(公告)日：2012-11-22

    申请号：US13474058

    申请日：2012-05-17

    申请人： Rastislav Levicky ,  Irina Belozerova ,  Ursula Koniges

    发明人： Rastislav Levicky ,  Irina Belozerova ,  Ursula Koniges

    IPC分类号： C40B30/04 ,  G01N27/26 ,  G01N21/64 ,  G01N25/04

    CPC分类号： G01N21/648 ,  C12Q1/6825 ,  G01N21/6428 ,  G01N27/021 ,  C12Q2525/301 ,  C12Q2527/107 ,  C12Q2563/113 ,  C12Q2565/1015

    摘要： Methods for analysis of interactions between nucleic acid-binding agents (BAs) and nucleic acids (NAs) by performance of nucleic acid denaturation assays on solid supports. Typically, BA is a small molecule less than 1000 g/gmol in molecular weight. The methods provide quantitative thermodynamic and kinetic analysis of BA-NA interaction; for example, in the form of free energies, enthalpies, and entropies of BA-NA binding in case of thermodynamic analysis, or in the form of rate constants and activation energies of BA-NA binding in the case of kinetic analysis. Examples of BAs of interest include transcription regulators and other NA-recognition molecules such as dyes and drug potentiators, DNA-targeted therapeutic agents including anticancer, antibiotic, antiviral, and antitrypanosomal compounds, carcinogens, and any other molecules whose interaction with DNA may, or is suspected to, lead to a biologically-relevant consequence. BA may bind to NA either through physical interactions or through formation of covalent adducts.

    摘要翻译： 通过在固体支持物上进行核酸变性测定来分析核酸结合剂（BAs）和核酸（NAs）之间相互作用的方法。 通常，BA是分子量小于1000g / gmol的小分子。 该方法提供了BA-NA相互作用的定量热力学和动力学分析。 在热力学分析的情况下，以BA-NA结合的自由能，焓和熵的形式，或在动力学分析的情况下为BA-NA结合的速率常数和活化能的形式。 感兴趣的BBA的实例包括转录调节剂和其它NA识别分子，例如染料和药物增强剂，DNA靶向治疗剂，包括抗癌剂，抗生素，抗病毒剂和抗蚂蚁胶原化合物，致癌物以及与DNA的相互作用的任何其它分子，或 怀疑是导致生物相关的后果。 BA可以通过物理相互作用或通过形成共价加合物与NA结合。

    Active CMOS biosensor chip for fluorescent-based detection
    7.
    发明申请
    Active CMOS biosensor chip for fluorescent-based detection 有权
    有源CMOS生物传感器芯片，用于荧光检测

    公开(公告)号：US20080037008A1

    公开(公告)日：2008-02-14

    申请号：US11800468

    申请日：2007-05-04

    申请人： Kenneth Shepard ,  Rastislav Levicky ,  George Patounakis

    发明人： Kenneth Shepard ,  Rastislav Levicky ,  George Patounakis

    IPC分类号： G01N21/00 ,  H01L27/14

    CPC分类号： H04N5/37452 ,  G01N21/6408 ,  G01N21/6428 ,  G01N21/6454 ,  H01L27/14621 ,  H01L27/14643

    摘要： An active CMOS biosensor chip for fluorescent-based detection is provided that enables time-gated, time-resolved fluorescence spectroscopy. In one embodiment, analytes are loaded with fluorophores that are bound to probe molecules immobilized on the surface of the chip. Photodiodes and other circuitry in the chip are used to measure the fluorescent intensity of the fluorophore at different times. These measurements are then averaged to generate a representation of the transient fluorescent decay response unique to the fluorophores. In addition to its low-cost, compact form, the biosensor chip provides capabilities beyond those of macroscopic instrumentation by enabling time-gated operation for background rejection, easing requirements on optical filters, and by characterizing fluorescence lifetime, allowing for a more detailed characterization of fluorophore labels and their environment. The biosensor chip can be used for a variety of applications including biological, medical, in-the-field applications, and fluorescent lifetime imaging applications.

    摘要翻译： 提供了用于基于荧光的检测的有源CMOS生物传感器芯片，其能够进行时间选择的时间分辨荧光光谱。 在一个实施方案中，分析物载有与固定在芯片表面上的探针分子结合的荧光团。 芯片中的光电二极管和其他电路用于在不同时间测量荧光团的荧光强度。 然后对这些测量进行平均以产生荧光团特有的瞬态荧光衰变响应的表示。 除了其低成本，紧凑的形式之外，生物传感器芯片通过实现背景抑制的时间门控操作，对光学滤波器的放宽要求以及通过表征荧光寿命来提供超出宏观仪器的功能，允许更详细的表征 荧光标签及其环境。 生物传感器芯片可用于各种应用，包括生物，医学，现场应用和荧光寿命成像应用。

    CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS
    8.
    发明申请
    CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS 有权
    用于核酸分析的电容性多元醇诊断

    公开(公告)号：US20120211375A1

    公开(公告)日：2012-08-23

    申请号：US13423228

    申请日：2012-03-18

    申请人： Rastislav Levicky ,  Napoleon Tercero ,  Kang Wang ,  Ping Gong ,  Kenneth Shepard

    发明人： Rastislav Levicky ,  Napoleon Tercero ,  Kang Wang ,  Ping Gong ,  Kenneth Shepard

    IPC分类号： G01N33/50 ,  G01N27/26

    CPC分类号： C12Q1/6825 ,  Y10T436/143333 ,  C12Q2525/113

    摘要： Use of Morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of Morpholino probes on gold supports are fabricated with methods similar to those employed with DNA, and are used to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for Morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a near-linear relationship, and demonstrates sensitivities (limits of quantification) in the pg mm−2 range.

    摘要翻译： 使用Morpholinos，一类不带电荷的DNA类似物，用于表面杂交应用。 使用类似于DNA使用的方法制备金支持物上的Morpholino探针的单层，并且用于与靶链有效地和序列特异性地杂交。 用电化学方法分析杂交诱导的界面电荷组织变化，并比较Morpholino和DNA探针单层。 通过与数值泊松 - 玻尔兹曼计算进行比较，可以确定和解释连接表面杂交态与界面电容的分子机制。 取决于由施加电位控制的移动离子的表面种群，对于杂交，阳性以及负电容反应（对比反转）是可能的。 表面电容与目标覆盖（目标/面积）的定量比较显示近似线性关系，并显示pg mm-2范围内的灵敏度（定量限度）。

    Active CMOS biosensor chip for fluorescent-based detection
    9.
    发明授权
    Active CMOS biosensor chip for fluorescent-based detection 有权
    有源CMOS生物传感器芯片，用于荧光检测

    公开(公告)号：US07738086B2

    公开(公告)日：2010-06-15

    申请号：US11800468

    申请日：2007-05-04

    申请人： Kenneth L. Shepard ,  Rastislav Levicky ,  George Patounakis

    发明人： Kenneth L. Shepard ,  Rastislav Levicky ,  George Patounakis

    IPC分类号： G01N21/00

    CPC分类号： H04N5/37452 ,  G01N21/6408 ,  G01N21/6428 ,  G01N21/6454 ,  H01L27/14621 ,  H01L27/14643

    摘要： An active CMOS biosensor chip for fluorescent-based detection is provided that enables time-gated, time-resolved fluorescence spectroscopy. In one embodiment, analytes are loaded with fluorophores that are bound to probe molecules immobilized on the surface of the chip. Photodiodes and other circuitry in the chip are used to measure the fluorescent intensity of the fluorophore at different times. These measurements are then averaged to generate a representation of the transient fluorescent decay response unique to the fluorophores. In addition to its low-cost, compact form, the biosensor chip provides capabilities beyond those of macroscopic instrumentation by enabling time-gated operation for background rejection, easing requirements on optical filters, and by characterizing fluorescence lifetime, allowing for a more detailed characterization of fluorophore labels and their environment. The biosensor chip can be used for a variety of applications including biological, medical, in-the-field applications, and fluorescent lifetime imaging applications.

    摘要翻译： 提供了用于基于荧光的检测的有源CMOS生物传感器芯片，其能够进行时间选择的时间分辨荧光光谱。 在一个实施方案中，分析物载有与固定在芯片表面上的探针分子结合的荧光团。 芯片中的光电二极管和其他电路用于在不同时间测量荧光团的荧光强度。 然后对这些测量进行平均以产生荧光团特有的瞬态荧光衰变响应的表示。 除了其低成本，紧凑的形式之外，生物传感器芯片通过实现背景抑制的时间门控操作，对光学滤波器的放宽要求以及通过表征荧光寿命来提供超出宏观仪器的功能，允许更详细的表征 荧光标签及其环境。 生物传感器芯片可用于各种应用，包括生物，医学，现场应用和荧光寿命成像应用。

    Active CMOS biosensor chip for fluorescent-based detection
    10.
    发明申请
    Active CMOS biosensor chip for fluorescent-based detection 审中-公开
    有源CMOS生物传感器芯片，用于荧光检测

    公开(公告)号：US20070097364A1

    公开(公告)日：2007-05-03

    申请号：US11431405

    申请日：2006-05-09

    申请人： Kenneth Shepard ,  Rastislav Levicky ,  George Patounakis

    发明人： Kenneth Shepard ,  Rastislav Levicky ,  George Patounakis

    IPC分类号： G01J3/30 ,  G01N21/64

    CPC分类号： G01J3/4406 ,  G01N21/6428 ,  G01N21/6454

    摘要： An active CMOS biosensor chip for fluorescent-based detection is provided that enables time-gated, time-resolved fluorescence spectroscopy. Analytes are loaded with fluorophores that are bound to probe molecules immobilized on the surface of the chip. Photodiodes and other circuitry in the chip are used to measure the fluorescent intensity of the fluorophore at different times. These measurements are then averaged to generate a representation of the transient fluorescent decay response unique to the fluorophores. In addition to its low-cost, compact form, the biosensor chip provides capabilities beyond those of macroscopic instrumentation by enabling time-gated operation for background rejection, easing requirements on optical filters, and by characterizing fluorescence lifetime, allowing for a more detailed characterization of fluorophore labels and their environment. The biosensor chip can be used for a variety of applications including biological, medical, in-the-field applications, and fluorescent lifetime imaging applications.

    摘要翻译： 提供了用于基于荧光的检测的有源CMOS生物传感器芯片，其能够进行时间选择的时间分辨荧光光谱。 分析物载有与固定在芯片表面上的探针分子结合的荧光团。 芯片中的光电二极管和其他电路用于在不同时间测量荧光团的荧光强度。 然后对这些测量进行平均以产生荧光团特有的瞬态荧光衰变响应的表示。 除了其低成本，紧凑的形式之外，生物传感器芯片通过实现背景抑制的时间门控操作，对光学滤波器的放宽要求以及通过表征荧光寿命来提供超出宏观仪器的功能，允许更详细的表征 荧光标签及其环境。 生物传感器芯片可用于各种应用，包括生物，医学，现场应用和荧光寿命成像应用。