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    • 5. 发明申请
    • CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS
    • 用于核酸分析的电容性多元醇诊断
    • WO2010088240A1
    • 2010-08-05
    • PCT/US2010/022178
    • 2010-01-27
    • POLYTECHNIC INSTITUTE OF NYULEVICKY, RastislavTERCERO, NapoleonWANG, KangGONG, PingSHEPARD, Kenneth
    • LEVICKY, RastislavTERCERO, NapoleonWANG, KangGONG, PingSHEPARD, Kenneth
    • C12Q1/68
    • C12Q1/6825Y10T436/143333C12Q2525/113
    • 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 organ ization are analyzed with electrochem ical 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.
    • 表面杂交,其中溶液中的核酸分子与固定在表面上的核酸对象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,来解决这些挑战的初步步骤。 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,并且显示出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,可以确定和解释将表面杂交态与界面电容连接的分子机制。 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)。
    • 6. 发明申请
    • ACTIVE CMOS SENSOR ARRAY FOR ELECTROCHEMICAL BIOMOLECULAR DETECTION
    • 用于电化学生物分子检测的有源CMOS传感器阵列
    • WO2009082706A1
    • 2009-07-02
    • PCT/US2008/087773
    • 2008-12-19
    • THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORKLEVINE, Peter, M.SHEPARD, Kenneth, L.GONG, PingRASTISLAV, Levicky
    • LEVINE, Peter, M.SHEPARD, Kenneth, L.GONG, PingRASTISLAV, Levicky
    • C12M3/00
    • G01N27/3277C12Q1/6825G01N27/48H01L23/5226H01L23/5283H01L23/53261
    • Electrochemical sensing of biomolecules eliminates the need for bulky optical instruments required in traditional fluorescence-based sensing assays. Integration of the sensor interface electrodes and active electrochemical detection circuitry on CMOS substrates miniaturizes the sensing platform, enhancing portability for point-of-care applications, while enabling high-throughput, highly-parallel analysis. One embodiment includes a four-by-four active sensor array for multiplexed electrochemical biomolecular detection in a standard 0.25-μm CMOS process. Integrated potentiostats, including control amplifiers and dual-slope ADCs, stimulate the electrochemical cell and detect the current flowing through on-chip gold electrodes at each sensor site resulting from biomolecular reactions occurring on the chip surface. Post-processing techniques for fabricating biologically-compatible surface-electrode arrays in CMOS that can withstand operation in harsh electrochemical environments are described. Demonstrations showing example operation of the active CMOS array for biomolecular detection include cyclic voltammetry of a reversible redox species, DNA probe density characterization, and quantitative and specific DNA hybridization detection in real time.
    • 生物分子的电化学感测消除了对传统基于荧光的感测测定所需的庞大的光学仪器的需要。 传感器接口电极和主动电化学检测电路在CMOS基板上的集成使传感平台小型化,增强了点对点应用的便携性,同时实现了高吞吐量,高度平行的分析。 一个实施例包括用于在标准0.25μmCMOS工艺中多路复用电化学生物分子检测的四乘四活性传感器阵列。 集成的恒电位仪,包括控制放大器和双斜率ADC,可激发电化学电池,并检测流经芯片表面上生物分子反应的每个传感器位置的片上金电极的电流。 描述了用于在CMOS中制造可承受恶劣电化学环境中的操作的生物相容的表面电极阵列的后处理技术。 显示用于生物分子检测的有源CMOS阵列的示例操作的演示包括可逆氧化还原物质的循环伏安法,DNA探针密度表征和实时定量和特异性DNA杂交检测。