公开(公告)号：US20160023239A1

公开(公告)日：2016-01-28

申请号：US14775947

申请日：2014-03-14

Applicant: PRESIDENT AND FELLOWS OF HARVARD COLLEGE

Inventor： Jene A. Golovchenko ,  Bo Lu ,  Aaron T. Kuan

IPC: B05D1/34

CPC classification number: B05D1/34 ,  B01D67/0034 ,  B01D67/0062 ,  B01D67/0065 ,  B01D69/02 ,  B01D2323/35 ,  B01D2325/02 ,  C25F3/00 ,  C25F3/14

Abstract: In a method for forming nanopores, two opposing surfaces of a membrane are exposed to an electrically conducting liquid environment. A nanopore nucleation voltage pulse, having a first nucleation pulse amplitude and duration, is applied between the two membrane surfaces, through the liquid environment. After applying the nanopore nucleation voltage pulse, the electrical conductance of the membrane is measured and compared to a first prespecified electrical conductance. Then at least one additional nanopore nucleation voltage pulse is applied between the two membrane surfaces, through the liquid environment, if the measured electrical conductance is no greater than the first prespecified electrical conductance.

Abstract translation: 在形成纳米孔的方法中，膜的两个相对表面暴露于导电液体环境。 具有第一成核脉冲幅度和持续时间的纳米孔成核电压脉冲通过液体环境施加在两个膜表面之间。 在应用纳米孔成核电压脉冲之后，测量膜的电导率并将其与第一预先指定的电导率进行比较。 然后，如果测量的电导率不大于第一预定电导，则通过液体环境，在两个膜表面之间施加至少一个另外的纳米孔成核电压脉冲。

公开(公告)号：US20150060277A1

公开(公告)日：2015-03-05

申请号：US14485628

申请日：2014-09-12

Applicant: President and Fellows of Harvard College ,  Peking University

Inventor： Jene A. Golovchenko ,  Bo Lu ,  David P. Hoogerheide ,  Dapeng Yu ,  Qing Zhao

IPC: G01N27/447 ,  G01N27/453

CPC classification number: G01N27/44765 ,  B01L3/502707 ,  B01L3/50273 ,  B01L3/502761 ,  B01L2200/0663 ,  B01L2300/0896 ,  B01L2400/0415 ,  B01L2400/0487 ,  C12Q1/6869 ,  G01N27/44791 ,  G01N27/453 ,  G01N33/48721 ,  C12Q2523/303 ,  C12Q2527/109 ,  C12Q2563/116 ,  C12Q2565/631

Abstract: There is provided a nanopore system including a nanopore in a support structure. A first reservoir is in fluidic connection with the nanopore and a second reservoir is in fluidic connection with the nanopore. The support structure separates the first and second reservoirs. A pressure source is connected to one of the first and second reservoirs to apply an external pressure to one of the first and second reservoirs. A voltage source is connected between the first and second reservoirs to apply a voltage bias between the first and second reservoirs, across the nanopore. This system enables a method for analysis of species in solution, wherein there is provided to the nanopore a fluidic solution that includes a species for translocation through the nanopore, with an external pressure applied to the species in fluidic solution and a voltage bias applied across the nanopore.

Abstract translation: 提供了一种纳米孔系统，其包括在支撑结构中的纳米孔。 第一储存器与纳米孔流体连接，并且第二储存器与纳米孔流体连接。 支撑结构分离第一和第二储存器。 压力源连接到第一和第二储存器中的一个，以向第一和第二储存器之一施加外部压力。 电压源连接在第一和第二储存器之间，以在第一和第二储存器之间穿过纳米孔施加电压偏置。 该系统使得能够分析溶液中的物质的方法，其中向纳米孔提供流体溶液，该流体溶液包括通过纳米孔易位的物质，其中施加到流体溶液中的物质的外部压力和施加在流体溶液中的电压偏置 纳米孔。

公开(公告)号：US20130313112A1

公开(公告)日：2013-11-28

申请号：US13962141

申请日：2013-08-08

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. DENISON ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

公开(公告)号：US20130270115A1

公开(公告)日：2013-10-17

申请号：US13852606

申请日：2013-03-28

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. DENISON ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

Abstract translation: 本发明涉及通过提供含有足以允许单链单体通过单链多核酸而不是双链多核酸的尺寸的通道的表面来表征目标多核酸的方法; 在表面提供杂交靶标多核酸的来源; 诱导目标多核酸通过通道，由此目标多核酸在其通过之前经历碱基对分离（熔融）; 并且随着靶多核酸相对于通道产生适合于确定目标多核苷酸的单体依赖特性的数据，随时间进行一个或多个测量。

公开(公告)号：US12140587B2

公开(公告)日：2024-11-12

申请号：US17319254

申请日：2021-05-13

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Stephen Jordan Fleming ,  Jene A. Golovchenko

IPC: G01N33/487 ,  B01D71/02 ,  B01D71/74 ,  C12Q1/6869 ,  G01N27/40 ,  B82Y35/00

Abstract: A nanopore system provided herein includes first and second fluidic reservoirs in fluidic communication with a nanopore forming a fluidic path between the reservoirs. An enzyme clamp, provided in the first fluidic reservoir, abuts the nanopore and is reversibly bound to a sequential plurality of polymer subunits of a target polymer molecule in ionic solution. The clamp has an outer clamp diameter that is greater than the nanopore diameter. An electrical circuit includes an electrode in each of the reservoirs for applying a voltage bias across the nanopore. A pulse generator is connected in the electrical circuit to apply control pulses across the nanopore to step the clamp along sequential polymer subunits of the target polymer molecule. The system includes no fuel or source of fuel for the clamp. A controller is connected in the electrical circuit for controlling the collection of electrical indications of polymer subunits.

公开(公告)号：US11639922B2

公开(公告)日：2023-05-02

申请号：US16629644

申请日：2018-07-17

Applicant: PRESIDENT AND FELLOWS OF HARVARD COLLEGE

Inventor： Lene V. Hau ,  Jene A. Golovchenko

IPC: G01N33/487

Abstract: Systems and methods are provided for characterizing shuttle capture events in a nanopore sensor. The method first collects time-dependent current blockage signatures for at least one bias voltage. The method then identifies each signature as corresponding to a permanent or transient event. The method then generates a protein dynamics landscape (PDL) for the transient event signatures. The PDL comprises a set of histograms of nanopore current data and characterizes current through the nanopore during shuttle capture events. The method can then comprise identifying an entrance level blockage value based on the permanent event signatures. Permanent event captures can be determined by time duration which is larger than a certain threshold time value. Applying a between the fluidic chambers above a threshold voltage level can be used to control that the vast majority of events are permanent.

公开(公告)号：US11567061B2

公开(公告)日：2023-01-31

申请号：US16629640

申请日：2018-07-17

Applicant: PRESIDENT AND FELLOWS OF HARVARD COLLEGE ,  UNIVERSITY OF MASSACHUSETTS

Inventor： Lene V. Hau ,  Jene A. Golovchenko ,  Min Chen

IPC: G01N33/487 ,  G01N27/447 ,  G01N33/543

Abstract: Systems and methods are provided for trapping and electrically monitoring molecules in a nanopore sensor. The nanopore sensor comprises a support structure with a first and a second fluidic chamber, at least one nanopore fluidically connected to the two chambers, and a protein shuttle. The protein shuttle comprises an electrically charged protein molecule, such as Avidin. The nanopore can be a Clytosolin A. A method can comprise applying a voltage across the nanopores to draw protein shuttles towards the nanopores. The ionic current through each or all of the nanopores can be concurrently measured. Based on the measured ionic current, blockage events can be detected. Each blockage event indicates a capture of a protein shuttle by at least one nanopore. Each blockage event can be detected through a change of the total ionic current flow or a change in the ionic current flow for a particular nanopore.

公开(公告)号：US20160139079A1

公开(公告)日：2016-05-19

申请号：US15004248

申请日：2016-01-22

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Jene A. Golovchenko

IPC: G01N27/447

CPC classification number: G01N27/44791 ,  B82Y15/00 ,  C12Q1/6869 ,  G01N27/44743 ,  G01N33/48721 ,  Y10S977/762 ,  C12Q2565/631

Abstract: Provided is a solid state support structure including an aperture having a molecular entrance and a molecular exit. A first reservoir is in fluidic communication with the molecular entrance of the aperture and contains a molecule-bearing liquid solution. A second reservoir is in fluidic communication with the molecular exit of the aperture for containing a molecule-bearing liquid solution. A first liquid channel is connected to the first reservoir within less than about 300 microns of the aperture in the support structure and includes molecule-bearing liquid solution for delivery to the first reservoir. A second liquid channel is connected to the second reservoir for accepting molecule-bearing liquid solution from the second reservoir. An electrical connection between the first reservoir and the second reservoir imposes an electrical bias between the first reservoir and the second reservoir for driving the molecule-bearing liquid solution through the aperture in the solid state support structure.

Abstract translation: 提供了一种固体支持结构，其包括具有分子入口和分子出口的孔。 第一储存器与孔的分子入口流体连通并且包含分子承载液体溶液。 第二储存器与用于容纳分子的液体溶液的孔的分子出口流体连通。 第一液体通道在支撑结构中的孔的小于约300微米内连接到第一储存器，并且包括用于输送到第一储存器的分子承载液体溶液。 第二液体通道连接到第二储存器，用于接收来自第二储存器的分子承载液体溶液。 第一储存器和第二储存器之间的电连接在第一储存器和第二储存器之间施加电偏压，用于驱动分子承载液体溶液穿过固态支撑结构中的孔。

公开(公告)号：US20210278392A1

公开(公告)日：2021-09-09

申请号：US17319254

申请日：2021-05-13

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Stephen Jordan Fleming ,  Jene A. Golovchenko

IPC: G01N33/487 ,  B01D71/02 ,  B01D71/74 ,  C12Q1/6869 ,  G01N27/40

Abstract: A nanopore system provided herein includes first and second fluidic reservoirs in fluidic communication with a nanopore forming a fluidic path between the reservoirs. An enzyme clamp, provided in the first fluidic reservoir, abuts the nanopore and is reversibly bound to a sequential plurality of polymer subunits of a target polymer molecule in ionic solution. The clamp has an outer clamp diameter that is greater than the nanopore diameter. An electrical circuit includes an electrode in each of the reservoirs for applying a voltage bias across the nanopore. A pulse generator is connected in the electrical circuit to apply control pulses across the nanopore to step the clamp along sequential polymer subunits of the target polymer molecule. The system includes no fuel or source of fuel for the clamp. A controller is connected in the electrical circuit for controlling the collection of electrical indications of polymer subunits.

公开(公告)号：US09046483B2

公开(公告)日：2015-06-02

申请号：US13962141

申请日：2013-08-08

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. Denison ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447 ,  C12Q1/68 ,  G01N33/487

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

Abstract translation: 本发明涉及通过提供含有足以允许单链单体通过单链多核酸而不是双链多核酸的尺寸的通道的表面来表征目标多核酸的方法; 在表面提供杂交靶标多核酸的来源; 诱导目标多核酸通过通道，由此目标多核酸在其通过之前经历碱基对分离（熔融）; 并且随着靶多核酸相对于通道产生适合于确定目标多核苷酸的单体依赖特性的数据，随时间进行一个或多个测量。