Bioluminescent bioreporter integrated circuit

• US 6,117,643 A
• Filed: 11/25/1997
• Issued: 09/12/2000
• Est. Priority Date: 11/25/1997
• Status: Expired due to Term

• Current Assignee: Lockheed Martin Energy Research Corporation (Martin Marietta Corporation), The University Of Tennessee Research Corporation (University of Tennessee)
• Original Assignee: The University Of Tennessee Research Corporation (University of Tennessee), UT-Battelle LLC
• Inventors: Simpson, Michael L., Paulus, Michael J., Sayler, Gary S.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/978,439
• Time in Patent Office: 1,022 Days
• Field of Search: 422/55, 422/57, 422/58, 422/82.01, 422/82.05, 422/82.06, 422/82.07, 422/82.08, 435/6, 435/7.32, 435/287.1, 435/287.2, 435/288.7, 435/808, 435/7.1, 436/518, 436/524, 436/525, 436/531, 436/805
• US Class Current: 435/7.1
• CPC Class Codes: A61B 5/0031 Implanted circuitry
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: C12Q 1/66 involving luciferase
• CPC Class Codes: G01N 21/763 Bioluminescence
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 435/849 Escherichia coli
• CPC Class Codes: Y10S 435/879 Salmonella
• CPC Class Codes: Y10S 435/909 Vibrio
• CPC Class Codes: Y10S 436/805 Optical property

Abstract

Disclosed are monolithic bioelectronic devices comprising a bioreporter and an OASIC. These bioluminescent bioreporter integrated circuit are useful in detecting substances such as pollutants, explosives, and heavy-metals residing in inhospitable areas such as groundwater, industrial process vessels, and battlefields. Also disclosed are methods and apparatus for environmental pollutant detection, oil exploration, drug discovery, industrial process control, and hazardous chemical monitoring.

Claims

1. An apparatus for detecting the concentration of a selected substance, comprising:

(a) a substrate;

(b) a bioreporter capable of metabolizing or interacting with a selected substance to emit light;

(c) a selectively permeable container affixed to said substrate capable of holding said bioreporter; and

(d) an integrated circuit on said substrate, including a phototransducer operative to generate a signal in response to said light.

2. The apparatus of claim 1 wherein the signal of step (d) indicates the presence of said substance.

3. The apparatus of claim 2, wherein said bioresistant/biocompatible material comprises silicon nitride.

4. The apparatus of claim 1, further comprising a layer of bioresistant and biocompatible material between said substrate and said container.

5. The apparatus of claim 1, wherein said integrated circuit is a Complementary Metal Oxide Semiconductor (CMOS) integrated circuit.

6. The apparatus of claim 1, wherein said phototransducer comprises a photodiode.

7. The apparatus of claim 1, wherein said integrated circuit further comprises a current to frequency converter and a digital counter.

8. The apparatus of claim 1, wherein said integrated circuit further comprises a photodiode and a current to frequency converter.

9. The apparatus of claim 1, wherein said integrated circuit further comprises a transmitter.

10. The apparatus of claim 9, wherein said transmitter is wireless.

11. The apparatus of claim 9, further comprising a receiver capable of receiving transmissions from said transmitter.

12. The apparatus of claim 11, wherein transmissions comprise digital data.

13. The apparatus of claim 1, further comprising a fluid and nutrient reservoir and a microfluidic pump on said substrate.

14. The apparatus of claim 1, wherein said bioreporter is selected from a group consisting of a genetically-engineered yeast, bacterium, fungal, animal and plant cell.

15. The apparatus of claim 1, wherein said selectively permeable container comprises a polymer matrix.

16. The apparatus of claim 15 wherein said polymer matrix is capable of allowing gas or fluid to reach said bioreporter.

17. The apparatus of claim 15, wherein said polymer matrix is optically clear.

18. The apparatus of claim 1, wherein said integrated circuit further comprises a global positioning system.

19. A monolithic bioelectronic device for detecting a substance, which comprises:

(a) a bioreporter atop a substrate on an integrated circuit, said bioreporter being capable of metabolizing the substance and emitting light consequent to such metabolism when in contact with said substance; and

,

(b) a sensor closely positioned to said integrated circuit to generate an electrical signal in response to receiving the emitted light.

20. The monolithic bioelectronic device of claim 19, wherein the bioreporter comprises a nucleic-acid segment encoding a bioluminescent marker polypeptide expressed in the presence of the substance.

21. The monolithic bioelectronic device of claim 20, wherein the sequence of the nucleic-acid segment encodes a luminescent reporter molecule.

22. The monolithic bioelectronic device of claim 19, further comprising a transparent, bioresistant and biocompatible separator positioned between the bioreporter and the sensor.

23. The monolithic bioelectronic device of claim 19, further comprising a polymer matrix encasing the bioreporter and enabling contact between the substance and the bioreporter.

24. The monolithic bioelectronic device of claim 23, wherein the polymer matrix is permeable to the substance.

25. The monolithic bioelectronic device of claim 19, wherein the bioreporter is identified as a microorganism.

26. The monolithic bioelectronic device of claim 25, which further comprises a source of water and nutrients suitable for sustaining the bioreporter.

27. Apparatus for detecting a substance which comprises:

(a) an integrated circuit including a phototransducer that produces a signal in response to light;

(b) a bioreporter capable of metabolizing the substance and emitting light consequent to such metabolism, wherein said reporter is in contact with said substance; and

(c) a transparent, biocompatible, and bioresistant separator positioned between the phototransducer and the bioreporter.

28. Apparatus as defined in claim 27, wherein the bioreporter comprises a nucleic-acid sequence encoding a lux gene, wherein said gene expresses a light-emitting polypeptide concurrently with the metabolism of said substance.

29. Apparatus as defined in claim 27 which further comprises a plastic matrix encasing the bioreporter and enabling contact between the substance and the bioreporter.

30. Apparatus as defined in claim 29 wherein the plastic matrix is permeable to the substance.

31. Apparatus as defined in claim 30 wherein the substance comprises a fluid.

32. Apparatus as defined in claim 27 in which the bioreporter comprises bacteria.