High throughput biochemical detection using single molecule fingerprinting arrays

Patent metadata

Field	Value
Publication number	US-9551682-B2
Application number	US-201213538346-A
Country	US
Kind code	B2
Filing date	Jun 29, 2012
Priority date	Jun 29, 2012
Publication date	Jan 24, 2017
Grant date	Jan 24, 2017

Abstract

Various embodiments provide devices, methods, and systems for high throughput biomolecule detection using transducer arrays. In one embodiment, a transducer array made up of transducer elements may be used to detect byproducts from chemical reactions that involve redox genic tags. Each transducer element may include at least a reaction chamber and a fingerprinting region, configured to flow a fluid from the reaction chamber through the fingerprinting region. The reaction chamber can include a molecule attachment region and the fingerprinting region can include at least one set of electrodes separated by a nanogap for conducting redox cycling reactions. In embodiments, by flowing the chamber content obtained from a reaction of a latent redox tagged probe molecule, a catalyst, and a target molecule in the reaction chamber through the fingerprinting region, the redox cycling reactions can be detected to identify redox-tagged biomolecules.

First claim

What is claimed is:

1. A method comprising:
obtaining a transducer array comprising a plurality of transducer elements, each transducer element comprising at least a reaction chamber and a fingerprinting region, wherein the fingerprinting region comprises at least one electrode set comprising a first electrode separated from a second electrode by a nanogap to enable a redox cycling reaction;
providing a latent redox tagged probe molecule, a catalyst, and a target molecule in the reaction chamber of at least one transducer element; and
flowing a chamber content from the reaction chamber and through the fingerprinting region to detect the redox cycling reaction in the nanogap between the first and the second electrodes that are biased to identify a corresponding redox-tag;
wherein the providing a latent redox tagged probe molecule, a catalyst, and a target molecule in the reaction chamber comprises providing (a) redox-tagged amino acids, a protease, and a peptide linker site in the reaction chamber or (b) an enzyme probe with latent redox tag, and a peptide target in the reaction chamber.

2. The method of claim 1 , wherein the corresponding redox-tag is directly or indirectly released from the probe molecule in the chamber content.

3. The method of claim 1 , wherein the redox cycling reaction detects an enzyme activity, a cell adhesion, a protein binding, and combinations thereof.

4. The method of claim 1 , wherein the transducer array is coupled with a readout circuit to read and interpret a signal from the redox cycling reaction.

5. The method of claim 1 , wherein the transducer array is coupled with a readout circuit to individually or in groups address one or more transducer elements of the plurality of transducer elements in the transducer array.

6. The method of claim 1 , further comprising changing an electrical bias on the first and the second electrodes to monitor a second redox cycling reaction.

7. The method of claim 1 , further comprising applying an electrical bias on the at least one electrode set of each of the one or more transducer elements of the plurality of transducer elements in the transducer array to detect a redox cycling reaction in parallel in the one or more transducer elements.

8. The method of claim 1 , wherein flowing the chamber content comprises using an active drift mechanism to unidirectionally direct the flow of the chamber contents.

9. The method of claim 1 , wherein τ cycle <τ measurement ≦τ exit <τ generation , and wherein τ generation is an average time for an incorporation event, τ exit is an average time for a molecule in the reaction chamber to exit the fingerprinting region, τ measurement is a time scale for electronically measuring current of the first and second electrodes, and τ cycle is an average time for the redox cycling between electrodes.

10. The method of claim 1 , further comprising simultaneously or sequentially flowing the chamber content from one or more reaction chambers of the transducer array.

11. The method of claim 1 , further comprising:
flowing the chamber content from the reaction chamber and through the fingerprinting region to detect the redox cycling reaction in the nanogap between the first and the second electrodes that are appropriately biased to identify a corresponding redox-tag.

12. The method of claim 1 , wherein the fingerprinting region comprises at least four electrode sets further comprising:
electrically biasing one or more of the at least four electrode sets in at least one transducer element;
wherein the transducer array is coupled with a readout circuit for implementing subtraction of the first and second electrode currents to reinforce anticorrelated redox signal, independently acquiring first and second electrode signals to reduce impact of amplifier noises by cross-correlation signal processing, using a switched capacitor implementation of a pair of transimpedance amplifiers with two separate outputs, or a combination thereof.

13. The method of claim 12 , wherein providing a latent redox tagged probe molecule, a catalyst, and a target molecule in the reaction chamber comprises providing a single polymerase, a DNA fragment to be sequenced, and redox-tagged nucleotides in the reaction chamber.

14. The method of claim 13 , further comprising determining sequence information for the DNA fragment to be sequenced based on the identified redox-tagged nucleotides.

15. The method of claim 13 , further comprising monitoring a signal from the redox cycling reaction as a function of time to determine a sequence of the DNA fragment to be sequenced.

16. The method of claim 13 , further comprising detecting or reading a DNA fragment of about 1-100 kilo bases long.

17. The method of claim 13 , wherein providing a single polymerase, a DNA fragment to be sequenced, and redox-tagged nucleotides in the reaction chamber comprises immobilizing the single polymerase on surface of the reaction chamber, the reaction chamber comprising the DNA fragment to be sequenced; and supplying the redox-tagged nucleotides to the reaction chamber.

18. The method of claim 12 , wherein the transducer array is coupled with a readout circuit to read and interpret a signal from the redox cycling reaction.

19. The method of claim 12 , wherein the redox cycling reaction is in situ detected in real-time in the fingerprinting region, upon a complimentary base incorporation in the reaction chamber.

20. The method of claim 12 , further comprising changing an electrical bias on each electrode set to monitor a corresponding redox cycling reaction.

21. The method of claim 12 , further comprising detecting the redox cycling reaction in parallel in each of one or more transducer elements selected from the plurality of transducer elements.

22. The method of claim 12 , wherein flowing the chamber content comprises using an active drift mechanism to unidirectionally direct the flow of the chamber contents.

23. The method of claim 12 , wherein τ cycle <τ measurement ≦τ exit <τ generation , and wherein τ generation is an average time for an incorporation event, τ exit is an average time for a molecule in the reaction chamber to exit the fingerprinting region, τ measurement is a time scale for electronically measuring current of the first and second electrodes, and τ cycle is an average time for the redox cycling between electrodes.

24. The method of claim 12 , wherein the redox-tagged nucleotides comprise at least four redox-tagged nucleotides with four redox tags respectively labeling four different nucleotides.

25. The method of claim 12 , further comprising conducting a whole genome sequencing by combining information from each transducer element of an array comprising a number of transducer elements to generate a complete order and identity of every base in the genome.

26. The method of claim 12 , further comprising simultaneously or sequentially flowing the chamber content from one or more reaction chambers of the transducer array.

27. The method of claim 12 , further comprising:
flowing the chamber content from the reaction chamber and through the fingerprinting region to detect the redox cycling reaction in each nanogap of the electrically biased one or more electrode sets to identify corresponding redox-tagged nucleotides.

Assignees

• Elibol Oguz H
• Credo Grace M
• Su Xing
• Varma Madoo
• Daniels Jonathan S
• Hall Drew
• Li Handong
• Tayebi Noureddine
• Wu Kai
• Intel Corp

Inventors

• Elibol Oguz H
• Credo Grace M
• Su Xing
• Varma Madoo
• Daniels Jonathan S
• Hall Drew
• Li Handong
• Tayebi Noureddine
• Wu Kai

Classifications

• G01N27/4148

  Integrated circuits therefor, e.g. fabricated by CMOS processing · CPC title

• G01N27/4146

  involving nanosized elements, e.g. nanotubes, nanowires · CPC title

• G01N27/4145

  specially adapted for biomolecules, e.g. gate electrode with immobilised receptors · CPC title

• G01N27/3278 Primary

  involving nanosized elements, e.g. nanogaps or nanoparticles (nanopores G01N33/48721; magnetic beads G01N27/745) · CPC title

• G01N27/3277 Primary

  being a redox reaction, e.g. detection by cyclic voltammetry (voltammetry per se G01N27/42, G01N27/48) · CPC title