In vivo biosensor apparatus and method of use

• US 6,673,596 B1
• Filed: 12/02/1999
• Issued: 01/06/2004
• Est. Priority Date: 11/25/1997
• Status: Expired due to Fees

• 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., Ripp, Steven A., Sayler, Gary S., Applegate, Bruce M.
• Primary Examiner(s): Ponnaluri, Padmashri
• Assistant Examiner(s): TRAN, MY CHAU T
• Application Number:
  US09/454,071
• Time in Patent Office: 1,496 Days
• Field of Search: 436/518, 436/524, 436/525, 436/531, 436/805, 424/9.1, 424/93.21, 424/172.1, 424/152.1, 424/1.45, 435/8, 435/189, 435/336, 435/6, 435/7.1, 435/7.32, 435/287.1, 435/287.2, 435/288.7, 435/808, 514/3, 422/55, 422/57, 422/58, 422/82.01, 422/82.05, 422/82.06, 422/82.07, 422/82.08
• US Class Current: 435/288.7
• 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 bioluminescent bioreporter integrated circuit devices that detect selected analytes in fluids when implanted in the body of an animal. The device comprises a bioreporter that has been genetically engineered to contain a nucleic acid segment that comprises a cis-activating response element that is responsive to the selected substance operably linked to a gene encoding a bioluminescent reporter polypeptide. In preferred embodiments, the target analyte is glucose, glucagons, or insulin. Exposure of the bioreporter to the target substance causes the response element to up-regulate the nucleic acid sequence encoding the reporter polypeptide to produce a luminescent response that is detected and quantitated. In illustrative embodiments, the bioreporter device is encapsulated on an integrated circuit that is capable of detecting the emitted light, processing the resultant signal, and then remotely reporting the results. Also disclosed are controlled drug delivery systems capable of being directly or indirectly controlled by the detection device that provide drugs such as insulin to the animal in reponse to the amount of target analyte present in the body fluids.

Claims

1. An implantable monolithic bioelectronic device for detecting at least one analyte within the body of an animal, said device comprising:

an integrated circuit including at least one transducer for generating an electrical signal in response to light incident thereon,

a bioreporter, said bioreporter for emitting light when exposed to said analyte, said bioreporter positioned so that at least a portion of said emitted light reaches said transducer, and

a biocompatible housing surrounding said integrated circuit and said bioreporter, said biocompatible housing including a semi-permeable membrane region for allowing passage of said analyte to said bioreporter but restricting passage of said bioreporter out from said housing.

2. The implantable monolithic bioelectronic device of claim 1, wherein said bioreporter is disposed in a polymeric matrix, said polymer matrix for keeping said bioreporter in place over said integrated circuit.

3. The implantable monolithic bioelectronic device of claim 2, wherein said polymeric matrix comprises polyvinyl alcohol, poly-L-lysine, or alginate.

4. The implantable monolithic bioelectronic device of claim 2, wherein said polymeric matrix comprises a microporous, mesh-reinforced or filter-supported hydrogel.

5. The implantable monolithic bioelectronic device of claim 1, wherein said integrated circuit comprises a phototransducer.

6. The implantable monolithic bioelectronic device of claim 5, further comprising a transparent, biocompatible, bioresistant separator operably positioned between the phototransducer and the bioreporter.

7. The implantable monolithic bioelectronic device of claim 1, wherein said bioreporter comprises a plurality of genetically engineered eukaryotic or prokaryotic cells, “

wherein said genetically engineered cells has Lux gene, such that the Lux gene”

permits said cells to produce a bioluminescent reporter polypeptide in response to the presence of said analyte.

8. The implantable monolithic bioelectric device of claim 7, wherein said plurality of eukaryotic cells comprise mammalian cells.

9. The implantable monolithic bioelectronic device of claim 8, wherein said plurality of eukaryotic cells comprise islet β

-cells, immortal stem cells, or hepatic cells.

10. The implantable monolithic bioelectronic device of claim 9, wherein said plurality of eukaryotic cells comprise recombinant human immortal stem cells.

11. The implantable monolithic bioelectronic device of claim 7, wherein said plurality of cells comprise a nucleic acid segment that encodes a luciferase polypeptide or a green fluorescent protein that is produced by said cells in response to the presence of said analyte.

12. The implantable monolithic bioelectronic device of claim 1, wherein said analyte is glucose, glucagon or insulin.

13. The implantable monolithic bioelectronic device of claim 1, further comprising a source of nutrients for sustaining cells disposed inside said biocompatible housing.

14. The implantable monolithic bioelectronic device of claim 1, further comprising a wireless transmitter, said wireless transmitter disposed inside said biocompatible housing and communicably connected to said transducer.

15. The implantable monolithic bioelectronic device of claim 1, further comprising an antenna, said antenna disposed inside said biocompatible housing and communicably connected to said transducer.

16. The implantable monolithic bioelectronic device of claim 1, further comprising an implantable drug delivery pump capable of being controlled by said device and a drug reservoir for holding at least one drug, said pump disposed inside said biocompatible housing, said pump for delivering said drug to the body of said animal.

17. The implantable monolithic bioelectronic device of claim 1, wherein said bioreporter expresses said light-emitting polypeptide following the metabolism of said analyte by said bioreporter.

18. The implantable monolithic bioelectronic device of claim 6, wherein said bioresistant separator comprises silicon nitride or silicon dioxide.

19. The implantable monolithic bioelectronic device of claim 1, wherein said integrated circuit is a complementary metal oxide semiconductor (CMOS) integrated circuit.

20. The implantable monolithic bioelectronic device of claim 5, wherein said phototransducer comprises a photodiode.

21. The implantable monolithic bioelectronic device of claim 1, wherein said integrated circuit further comprises a photodiode and a current to frequency converter.

22. The implantable monolithic bioelectronic device of claim 1, wherein said integrated circuit further comprises a current to frequency converter and a digital counter.

23. The implantable monolithic bioelectronic device of claim 1, further comprising a wireless transmitter, said wireless transmitter disposed inside said biocompatible housing and communicably connected to said transducer.

24. The implantable monolithic bioelectronic device of claim 23, wherein said transmitter is for transmitting digital data.

25. An implantable controlled drug delivery system, comprising the device of claim 1, further comprising an implantable drug delivery pump and a drug reservoir disposed inside said biocompatible housing for providing at least one drug to said animal, said bioelectric device controlling a level of said drug provided to said animal based on a measured concentration of said analyte.

26. A kit for the detection of an analyte comprising the device of claim 1 and instructions for using said device.

27. The kit of claim 26, further comprising a standardized reference solution.