Apparatus and method for biopolymer indentification during translocation through a nanopore

• US 20040144658A1
• Filed: 01/27/2003
• Published: 07/29/2004
• Est. Priority Date: 01/27/2003
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Flory, Curt A.
• Granted Patent: US 7,410,564 B2
• Time in Patent Office: Days
• US Class Current: 205/777.5
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 15/1245 Devices using more than one aperture
• CPC Class Codes: G01N 15/134 Devices using two or more apertures
• CPC Class Codes: G01N 33/48721 Investigating individual macromolecules, e.g. by translocation through nanopores Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68

Abstract

The present invention provides an apparatus and method for identifying and sequencing a biopolymer translocating a nanopore. The apparatus of the present invention provides a first electrode, a second electrode and a potential means for applying a bias ramping potential across the electrodes to produce resonant tunneling of current carriers between the two electrodes. As the bias potential is ramped across the electrodes the increase in tunneling current occurs as the carrier energy sequentially matches the conduction band energies of the translocating biopolymer. This technique allows for real-time identification and sequencing of a biopolymer as the band energy spectra of the individual portions of the bipolymer are recorded, differentiated and identified. A method for identifying the biopolymer is also disclosed.

Claims

1. An apparatus for detecting a biopolymer in a nanopore, comprising:

(a) a first ring electrode;

(b) a second ring electrode adjacent to said first ring electrode;

(c) a nanopore adjacent to said first ring electrode and said second ring electrode and positioned to allow said biopolymer to be positioned in said first ring electrode and said second ring electrode; and

(d) potential means for electrically connecting said first ring electrode and said second ring electrode for applying a ramping potential from said first ring electrode, through a portion of said biopolymer in said nanopore, to said second ring electrode to produce a signal indicative of said portion of said biopolymer.

2. The apparatus as recited in claim 1, further comprising a substrate for positioning said first electrode and said second electrode.

3. The apparatus as recited in claim 1, further comprising at least a first substrate for positioning said first electrode.

4. The apparatus as recited in claim 1, further comprising at least a second substrate for positioning said second electrode.

5. The apparatus of claim 1, further comprising at least a first substrate for positioning a nanopore.

6. The apparatus of claim 1, further comprising a means for signal detection for detecting said signal produced from said portion of said biopolymer.

7. The apparatus of claim 1, wherein said biopolymer is a charged polymer.

8. The apparatus of claim 6, wherein said biopolymer is selected from the group consisting of carbohydrates, proteins, nucleic acids, lipids, glycans, polynucleotides, proteoglycans and polypeptides.

9. A method for identifying a biopolymer translocating through a nanopore, comprising:

(a) applying a ramping electrical current from a first ring electrode through a portion of said biopolymer to a second ring electrode to identify said portion of said biopolymer positioned in said nanopore.

10. A method as recited in claim 9, wherein said electrical current is a tunneling current with an energy level that matches at least one conduction band energy of a portion of said biopolymer.

11. A method as recited in claim 10, wherein said tunneling current is on resonance with said conduction band energies of a portion of said biopolymer.

12. A method as recited in claim 9, further comprising translocating said biopolymer through said nanopore to identify each of said translocating portions of said biopolymer.

13. An apparatus for detecting a bipolymer translocating a nanopore, comprising:

(a) a first electrode having a first nanopore;

(b) a second electrode adjacent to said first electrode having a second nanopore wherein said first nanopore of said first electrode is positioned with said second nanopore of said second electrode so that said bipolymer may translocate through said first nanopore and said second nanopore; and

(c) potential means for electrically connecting said first electrode and said second electrode for applying a ramping potential from said first electrode through a portion of said biopolymer to said second electrode to produce a detectable signal indicative of a portion of said biopolymer translocating said first nanopore and said second nanopore.

14. An apparatus as recited in claim 13, wherein the center point of said nanopore of said first electrode is positioned coaxially with the center point of said nanopore of said second electrode.

15. An apparatus as recited in claim 14, wherein said first electrode is positioned above said second electrode.

16. An apparatus as recited in claim 13, further comprising a substrate for positioning said first electrode and said second electrode.

17. An apparatus as recited in claim 13, further comprising at least a first substrate for positioning said first electrode.

18. An apparatus as recited in claim 13, further comprising at least a second substrate for positioning said second electrode.

19. An apparatus for detecting a portion of a biopolymer tanslocating a nanopore, comprising:

(a) a first electrode;

(b) a second electrode spaced from said first electrode to define a nanopore between said first electrode and said second electrode, said nanopore designed for receiving a translocating biopolymer, said first electrode being in electrical connection with said second electrode; and

(c) potential means for electrically connecting said first electrode and said second electrode for applying a ramping bias potential from said first electrode, across said biopolymer to said second electrode to produce a signal indicative of a portion of said biopolymer translocating said nanopore.

20. An apparatus as recited in claim 19, wherein said biopolymer is translocated in a stepwise fashion through said nanopore defined between said first electrode and said second electrode.

21. A method for identifying a biopolymer in a nanopore defined between a first and second electrode, comprising:

applying a ramping potential from a first electrode through a portion of a biopolymer to a second electrode to scan energy levels until the energy levels of said first electrode, said biopolymer and said second electrode are on resonance, said ramping potential to produce a detectable signal to identify said portion of said biopolymer positioned in said nanopore.