ENZYME-LOGIC BIOSENSING

• US 20130065257A1
• Filed: 03/16/2011
• Published: 03/14/2013
• Est. Priority Date: 03/16/2010
• Status: Abandoned Application

• Current Assignee: Clarkson University (Union Graduate College), Regents of the University of California (University of California)
• Original Assignee: Jan Halamek
• Inventors: Wang, Joseph, Windmiller, Joshua Ray, Santhosh, Padmanabhan, Chuang, Min-Chieh, Katz, Evgeny, Halmek, Jan, Bocharova, Vera, Pita, Marcos, Privman, Vladimir
• Application Number:
  US13/635,396
• Publication Number:
  US 20130065257A1
• Time in Patent Office: Days
• US Class Current: 435/7.92
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: G01N 21/31 Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry G01N21/72 takes precedence
• CPC Class Codes: G01N 2333/902 Oxidoreductases (1.)
• CPC Class Codes: G01N 2333/90241 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: G01N 2333/904 acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
• CPC Class Codes: G01N 2333/908 acting on hydrogen peroxide as acceptor (1.11)
• CPC Class Codes: G01N 2333/91188 transferring nitrogenous groups (2.6)
• CPC Class Codes: G01N 2333/91205 Phosphotransferases in general
• CPC Class Codes: G01N 2333/988 Lyases (4.), e.g. aldolases, heparinase, enolases, fumarase
• CPC Class Codes: G01N 27/27 Association of two or more measuring systems or cells, each measuring a different parameter, where the measurement results may be either used independently, the systems or cells being physically associated, or combined to produce a value for a further parameter
• CPC Class Codes: G01N 27/327 Biochemical electrodes , e.g. electrical or mechanical details for in vitro measurements
• CPC Class Codes: G06N 3/002 Biomolecular computers, i.e. using biomolecules, proteins, cells using DNA G06N3/123; using neurons G06N3/061

Abstract

Techniques, apparatus and systems are disclosed for implementing enzyme-logic based diagnosis that uses patterns of multiple markers and biochemical processing of the signal information for reliably identifying cardiac abnormalities and providing a final digital binary answer. In one aspect, a biochemical logic sensing system includes a network of enzyme-biocatalyzed logic gates adapted to receive biomarker input signals and perform an enzyme-biocatalyzed reaction resembling a Boolean logic operation using the received biomarker input signals to generate an output signal of the enzyme-biocatalyzed reaction. A signal processing unit is connected to the network of enzyme-biocatalyzed logic gates. The signal processing unit processes the generated output signal of the enzyme-biocatalyzed reaction and generates a digital binary output having a value of zero or one. The generated digital binary output indicates a type of an injury.

Claims

1. A biochemical logic sensing system, comprising:

a network of enzyme-biocatalyzed logic gates adapted to receive biomarker inputs and perform an enzyme-biocatalyzed reaction resembling a Boolean logic operation using the received biomarker inputs to generate an output of the enzyme-biocatalyzed reaction; and

a signal processing unit connected to the network of enzyme-biocatalyzed logic gates, the signal processing unit to process the generated output of the enzyme-biocatalyzed reaction and generate a digital binary output having a value of zero or one,

wherein the generated digital binary output indicates an injury.

2. The biochemical logic sensing system of claim 1, further comprising:

a drug delivery device connected to the signal processing unit, the drug delivery device identifying a dosage of a drug to deliver based on the processed digital binary output.

3. The biochemical logic sensing system of claim 1, wherein the signal processing unit is adapted to generate the digital binary output having a value of 0 when the network of enzyme-biocatalyzed logic gates detects a physiologically normal concentration of the biomarker inputs;

and

wherein the signal processing unit is adapted to generate the digital binary output having a value of 1 when the network of enzyme-biocatalyzed logic gates detects an abnormal concentration of the biomarker input.

4. The biochemical logic sensing system of claim 1, wherein the biomarker inputs comprise glucose, lactate, norepinephrine (NE) and oxygen.

5. The biochemical logic sensing system of claim 4, wherein the signal processing unit is adapted to generate the digital binary output having a value of 1 when the network of enzyme-biocatalyzed logic gates detects at least one of:

an abnormal increase in glucose concentration associated with hemorrhagic shock (HS);

a higher than normal physiological concentration of lactate associated with HS and/or trauma brain injury (TBI);

a high concentration of NE associated with a traumatic injury; and

lack of oxygen associated with ischemic state and/or heart attack.

6. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates comprises at least one of an AND gate, an XOR gate, an OR gate, a NAND gate, a NOR gate and an IDENTITY gate.

7. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates are adapted to form a cascade arrangement.

8. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to form parallel-operating enzyme-catalyzed pathways producing outputs of two or more enzyme-biocatalyzed reactions in parallel.

9. The biochemical logic sensing system of claim 8, wherein the signal processing unit is adapted to:

generate two or more digital binary output in parallel based on the outputs of two or more enzyme-biocatalyzed reactions performed in parallel; and

identify a type of an injury based on at least one of the generated digital binary output signals.

10. The biochemical logic sensing system of claim 1, further comprising:

a monitoring device connected to the signal processing unit, the monitoring device adapted to monitor the output of the enzyme-biocatalyzed reaction.

11. The biochemical logic sensing system of claim 1, wherein the monitoring device comprises at least one of an optical monitor and an electrochemical monitor.

12. The biochemical logic sensing system of claim 1, wherein the biomarker inputs comprise lactate, norepinephrine and glucose;

and

the network of enzyme-biocatalyzed logic gates comprises at least one of an AND gate and an IDENTITY gate adapted to operate in concert using lactate oxidase, horseradish peroxidase and glucose dehydrogenase enzymes.

13. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process different patterns of the biomarker inputs to generate the output of the enzyme-biocatalyzed reaction comprising norepi-quinone and reduced dipotassium salt (NADH).

14. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs to provide diagnostic information on multiple conditions at a point-of-care.

15. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs to provide diagnostic information on a cardiovascular event.

16. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs comprising myeloperoxidase (MPO), lactate dehydrogenase (LDH) and creatine kinase (CK), and generate the output signals comprising reduced dipotassium salt (NADH) and oxidized redox mediator (Mox).

17. The biochemical logic sensing system of claim 1, wherein the signal processing unit is adapted to process the output of the enzyme-biocatalyzed reaction comprising reduced dipotassium salt (NADH) and oxidized redox mediator (Mox) to generate the digital binary output that indicates at least one of four possible symptoms comprising a healthy heart (0,0), unhealthy body tissue (1,0), a heart related illness different from Acute myocardial infarction (AMI) (0,1) and AMI (1,1).

18. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to analyze the biomarker inputs that are characteristic of at least one of liver injury (LI), soft tissue injury (STI) and abdominal trauma (ABT).

19. The biochemical logic sensing system of claim 1, wherein the signal processing unit is adapted to use the generated the digital binary output to produce an alert-type optical output signal in the form of YES-NO separated by a threshold value.

20. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates comprises at least one of AND, OR, NAND, NOR, and XOR gates.

21. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs comprising alanine transaminase (ALT) and lactate dehydrogenase (LDH) associated with liver injury.

22. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs comprising CK and LDH associated with soft tissue injury.

23. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs comprising enzyme LDH and its substrate Lac at elevated concentrations associated with abdominal trauma (ABT).

24. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process simultaneously different combinations of five biomarker inputs characteristic of traumatic brain injury (TBI) and soft tissue injury (STI).

25. The biochemical logic sensing system of claim 24, wherein three of the biomarker inputs comprise creatine kinase (CK), lactate dehydrogenase (LDH) and lactate (Lac) associated with physiological conditions characteristic of STI.

26. The biochemical logic sensing system of claim 24, wherein two of the biomarker inputs comprise enolase (EN) and glutamate (Glu) associated with the TBI diagnosis.

27. The biochemical logic sensing system of claim 24, wherein the network of enzyme-biocatalyzed logic gates is adapted to switch between an STI detection mode and a TBI detection mode.

28. The biochemical logic sensing system of claim 24, wherein the network of enzyme-biocatalyzed logic gates is adapted to process simultaneous presence of elevated levels of CK and LDH to trigger a positive diagnosis of STI.

29. The biochemical logic sensing system of claim 1, further comprising a disposable electrode to obtain a blood sample.

30. The biochemical logic sensing system of claim 1, wherein the disposable electrode comprises a flexible carbon screen-printed electrode (SPE).

31. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to assess the biomarker inputs comprising two or more selected from creatine kinase, lactate dehydrogenase, norepinephrine, glutamate, alanine transaminase, lactate, glucose, glutathione disulfide, and glutathione reductase to assess associated one of soft-tissue injury, traumatic brain injury, liver injury, abdominal trauma, hemorrhagic shock, and oxidative stress.

32. The biochemical logic sensing system of claim 1, wherein the signal processing unit is adapted to generate a digitally-encoded multi-bit binary word that represents an injury code.

33. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates are arranged in parallel and multiplexed.

34. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to form different arrangements with each enzyme-biocatalyzed logic gate receiving two of the biomarker inputs.

35. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates are adapted to process the biomarker inputs that comprise Glutathione disulfide (GSSG) and glucocorticoid receptor (GR) associated with oxidative stress.

36. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs comprising alanine transaminase (ALT) and lactate dehydrogenase (LDH) to generate NAD output signal;

and

wherein the biochemical logic sensing system further comprises a switchable polymer-modified electrode connected to the signal processing unit, the switchable polymer-modified electrode adapted convert the generated NAD output signal to a change in pH.

37. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates comprises:

a first AND logic gate activated by lactate dehydrogenase (LDH) and lactate—

jointly representing biomarker inputs for abdominal trauma (ABT); and

a second AND logic gate was activated by glutathione reductase (GR) and glutathione disulfide (GSSG) representative of oxidative stress when their concentrations are elevated.

38. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs comprising α

-amylase (Amy) and lactate dehydrogenase (LDH) to analyze radiation-caused tissue damage.

39. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs to detect the presence of at least one of explosive compounds and nerve agents.

40. The biochemical logic sensing system of claim 39, wherein the network of enzyme-biocatalyzed logic gates is adapted to process the biomarker inputs that comprises 2,4,6-trinitrotoluene (TNT) and paraoxon (PAX) to detection 2,4-dinitrotoluene (DNT) explosive and methyl-parathion (MPT) nerve agent.

41. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to utilize enzymes comprising zanitroreductase (NRd), horseradish peroxidase (HRP), acetylcholinesterase (AChE), and choline oxidase (ChOx).

42. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is adapted to simultaneously process biochemically coupled diabetes biomarker inputs comprising glucose and hemoglobin A1c (HbA1c) to generate the output of the enzyme-biocatalyzed reaction;

and

the signal processing unit is adapted to process the generated output of the enzyme-biocatalyzed reaction to generate the digital binary output indicative of diabetes.

43. The biochemical logic sensing system of claim 1, wherein the network of enzyme-biocatalyzed logic gates is disposed on a single use test strip.

44. The biochemical logic sensing system of claim 43, wherein the single use test strip comprises:

a reporter anti-HbA1c polyclonal antibody labeled with a horseradish peroxidase (HRP), which is deposited in a sampling zone patch; and

a capture anti-HbA1c analyte-specific polyclonal antibody immobilized on a surface of a detection zone together with glucose oxidase (GOx) and dried redox dye.

45. The biochemical logic sensing system of claim 44, wherein the single use test strip is adapted to:

receive a liquid sample comprising glucose onto the sample zone to solubilize the labeled reporter antibody, which binds to the HbA1c to for an analyte-antibody complex;

when the analyte-antibody complex flows with the liquid sample laterally along the surface of the test strip over the detection zone, the analyte-antibody complex binds to the capture antibody to accumulate at the capture zone of the test strip; and

the immobilized GOx converts glucose present in the sample to hydrogen peroxidse, which serves as a substrate for the HRP-labeled reporter antibody present on the HbA1c molecule.

48. The method of claim 1, comprising:

operating a monitoring device to analyze the output of the enzyme-biocatalyzed reaction.

49. The method of claim 1, comprising:

operating the network of enzyme-biocatalyzed logic gates to process the biomarker inputs that includes one or a combination of multiple biomarkers associated with a particular type of a medical condition.

50. The method of claim 1, comprising:

operating the network of enzyme-biocatalyzed logic gates to perform enzyme-biocatalyzed reactions in parallel and generate outputs of the enzyme-biocatalyzed reactions in parallel; and

operating the signal processing unit to generate digital binary outputs in parallel in response to the parallelly generated outputs of the enzyme-biocatalyzed reactions to detect multiple medical conditions.

51. The method of claim 1, comprising:

operating the signal processing unit to generate an injury code comprising a multi-bit digital word.

52. The method of claim 1, comprising:

operating the network of enzyme-biocatalyzed logic gates to process multiple biomarker inputs associated with a given medical condition.

53. The method of claim 1, comprising:

operating the network of enzyme-biocatalyzed logic gates to switch between multiple modes of operation.

46. A method of detecting a medical condition using a biochemical sensing system comprising:

operating a network of enzyme-biocatalyzed logic gates in a biochemical sensing system to perform an enzyme-biocatalyzed reaction resembling a Boolean logic operation on biomarker inputs to generate an output of the enzyme-biocatalyzed reaction;

operating a signal processing unit in the biochemical sensing system to generate a digital binary output based on the generated output of the enzyme-biocatalyzed reaction, wherein the digital binary output indicates presence or absence of a medical condition.

47. The method of claim 46, comprising:

operating a drug deliver device to determine a dosage of a drug to deliver based on the digital binary output from the signal processing unit.