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：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：US20250060354A1

公开(公告)日：2025-02-20

申请号：US18912894

申请日：2024-10-11

Applicant: President and Fellows Of Harvard College

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

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

Abstract: A method is provided for deterministically translocating through a nanopore a target polymer molecule of a nucleic acid polymer molecule or a protein polymer molecule. In the method, an enzyme clamp is reversibly bound to a plurality of sequential polymer subunits of the target polymer molecule. The target polymer molecule and the enzyme clamp are disposed at the nanopore. In the method, there is applied a pulse of force operative to deterministically advance the enzyme clamp along the target polymer molecule by no more than one polymer subunit. The pulse of force is then repeatedly applied to cause deterministic translocation of a sequential plurality of polymer subunits of the target polymer molecule through the nanopore.

公开(公告)号：US11035847B2

公开(公告)日：2021-06-15

申请号：US16022594

申请日：2018-06-28

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: In a method p for controlling translocation of a target polymer molecule through a nanopore, a clamp is reversibly bound to a sequential plurality of polymer subunits along the target polymer molecule length and the molecule and clamp are disposed in an ionic solution that is in fluidic communication with the nanopore. A constant translocation force is applied across the nanopore to induce travel of the target polymer molecule into the nanopore, until the clamp abuts the nanopore aperture and stops further travel of the target polymer molecule into the nanopore. Then a voltage control pulse is applied across the nanopore and/or a thermal control pulse is applied at the nanopore, with a pulse duration that steps the clamp along the target polymer molecule by no more than one polymer subunit in a direction opposite that of travel into the nanopore. No fuel is provided to the clamp.

公开(公告)号：US20190004029A1

公开(公告)日：2019-01-03

申请号：US16022594

申请日：2018-06-28

Applicant: President and Fellows of Harvard College

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

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

Abstract: In a method p for controlling translocation of a target polymer molecule through a nanopore, a clamp is reversibly bound to a sequential plurality of polymer subunits along the target polymer molecule length and the molecule and clamp are disposed in an ionic solution that is in fluidic communication with the nanopore. A constant translocation force is applied across the nanopore to induce travel of the target polymer molecule into the nanopore, until the clamp abuts the nanopore aperture and stops further travel of the target polymer molecule into the nanopore. Then a voltage control pulse is applied across the nanopore and/or a thermal control pulse is applied at the nanopore, with a pulse duration that steps the clamp along the target polymer molecule by no more than one polymer subunit in a direction opposite that of travel into the nanopore. No fuel is provided to the clamp.