Sensor circuit for controlling, detecting, and measuring a molecular complex

• US 10,724,987 B2
• Filed: 12/21/2015
• Issued: 07/28/2020
• Est. Priority Date: 02/27/2012
• Status: Active Grant

• Current Assignee: Roche Sequencing Solutions, Inc. (Roche Holding AG)
• Original Assignee: Roche Sequencing Solutions, Inc. (Roche Holding AG)
• Inventors: Deierling, Kevin, Chen, Roger J. A., Fullagar, David J.
• Primary Examiner(s): Eom, Robert J
• Application Number:
  US14/977,312
• Publication Number:
  US 20160266064A1
• Time in Patent Office: 1,681 Days
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• CPC Class Codes: G01N 27/4473 by electric means
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• 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
• CPC Class Codes: Y10T 436/11 Automated chemical analysis

Abstract

A device for controlling, detecting, and measuring a molecular complex is disclosed. The device comprises a common electrode. The device further comprises a plurality of measurement cells. Each measurement cell includes a cell electrode and an integrator electronically coupled to the cell electrode. The integrator measures the current flowing between the common electrode and the cell electrode. The device further comprises a plurality of analog-to-digital converters, wherein an integrator from the plurality of measurement cells is electrically coupled to one analog-to-digital converter of the plurality of analog-to-digital converters.

Claims

1. A device, the device comprising:

a precharging voltage source;

a first cell electrode;

a second electrode, the second electrode configured to apply a liquid potential to a liquid in contact with a nanopore positioned electrically between the first cell electrode and the second electrode;

a capacitor electronically coupled to the first cell electrode;

an electronic switch that selectively couples the capacitor to the precharging voltage source;

an analog-to-digital converter electrically coupled to the capacitor; and

a processor programmed to;

send a precharge signal to the electronic switch to close the electronic switch and electrically couple the capacitor to the precharging voltage source;

precharge the capacitor to a set voltage at a beginning of a measurement period;

send a signal to the electronic switch to open the electronic switch to decouple the capacitor from the precharging voltage source; and

measure a decay in voltage across the capacitor during the measurement period using the analog-to-digital converter.

2. The device of claim 1, further comprising a liquid chamber containing the liquid.

3. The device of claim 2, wherein

the second electrode supplies the liquid potential when the liquid chamber contains an electrolyte;

the first cell electrode supplies a variable electric potential when the liquid chamber contains the electrolyte, wherein a voltage between the second electrode and the first cell electrode equals the variable electric potential minus the liquid potential; and

the decay in voltage across the capacitor is a function of a resistance through the nanopore.

4. The device of claim 3, wherein,

the device is configured to detect a physical state and transpose the physical state from a plurality of possible physical states of a material in the liquid chamber, by modulating the voltage between the second electrode and the first cell electrode and measuring a voltage decay between the first cell electrode and the second electrode, and

the plurality of possible physical states comprises:

no formation of a lipid bilayer,

formation of a lipid bilayer,

insertion of a nanopore protein in a lipid bilayer, forming a nanopore, and

interaction of a molecular complex with the nanopore.

5. The device of claim 2, wherein the device is configured to retain at least one lipid, one nanopore protein, and one molecular complex in the liquid chamber.

6. The device of claim 5, wherein the device is configured to recognize a specific base type corresponding to the molecular complex in the liquid chamber by using the measurement of the decay in voltage across the capacitor.

7. The device of claim 1, further comprising

a buffering component that is electrically coupled to the analog-do-digital converter to buffer an output of the capacitor before being connected to the analog-do-digital converter.

8. The device of claim 1, wherein the analog-to-digital converter electrically coupled to the capacitor measures a first voltage at an output of the capacitor at the beginning of the measurement period and a second voltage at the output of the capacitor at an end of the measurement period, and wherein a difference of the second voltage and the first voltage corresponds to the measurement of the decay in voltage across the capacitor.

9. The device of claim 8, wherein the measurement period is set or predetermined and remains fixed between measurements.

10. The device of claim 1, wherein the analog-to-digital converter includes a comparator.

11. The device of claim 10, wherein the analog-to-digital converter includes a controlling analog input, and the controlling analog input controls a variable sample rate of a current flowing between the first cell electrode and the second electrode by varying a voltage at the controlling analog input.

12. The device of claim 11, wherein the variable sample rate increases when the current flowing between the first cell electrode and the second electrode increases.

13. The device of claim 11, wherein the varied voltage comprises a comparator threshold.

14. The device of claim 11, wherein the varied voltage comprises an initial voltage at an output of the capacitor.

15. A device, the device comprising:

a precharging voltage source;

a first cell electrode;

a second electrode, the second electrode configured to apply a liquid potential to a liquid in contact with a nanopore positioned electrically between the first cell electrode and the second electrode;

a capacitor electronically coupled to the first cell electrode;

an electronic switch that selectively couples the capacitor to the precharging voltage source;

an analog-to-digital converter electrically coupled to the capacitor; and

a processor programmed to;

send a precharge signal to the electronic switch to close the electronic switch and electrically couple the capacitor to the precharging voltage source;

precharge the capacitor to a set voltage at a beginning of a measurement period;

send a signal to the electronic switch to open the electronic switch to decouple the capacitor from the precharging voltage source; and

measure a decay in voltage across the capacitor during the measurement period using the analog-to-digital converter, wherein the measurement period is set or predetermined and remains fixed between measurements.

16. A device, the device comprising:

a precharging voltage source;

a first cell electrode;

a second electrode, the second electrode configured to apply a liquid potential to a liquid in contact with a nanopore positioned electrically between the first cell electrode and the second electrode;

a capacitor electronically coupled to the first cell electrode;

an electronic switch that selectively couples the capacitor to the precharging voltage source;

an analog-to-digital converter electrically coupled to the capacitor; and

a processor programmed to;

send a precharge signal to the electronic switch to close the electronic switch and electrically couple the capacitor to the precharging voltage source;

precharge the capacitor to a set voltage at a beginning of a measurement period;

send a signal to the electronic switch to open the electronic switch to decouple the capacitor from the precharging voltage source; and

measure a decay in voltage across the capacitor during the measurement period using the analog-to-digital converter, wherein the measurement period is determined based on a time for the measured decay in voltage across the capacitor to reach a fixed magnitude.