Bioluminescent bioreporter integrated circuit devices and methods for detecting estrogen

• US 7,090,992 B2
• Filed: 06/14/2005
• Issued: 08/15/2006
• Est. Priority Date: 11/25/1997
• Status: Expired due to Fees

• Current Assignee: UT-Battelle LLC
• Original Assignee: UT-Battelle LLC
• Inventors: Simpson, Michael L., Paulus, Michael J., Ripp, Steven A., Sayler, Gary S., Applegate, Bruce M.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US11/152,138
• Publication Number:
  US 20060014228A1
• Time in Patent Office: 427 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, 435/287.1, 435/287.2, 435/288.7, 435/808, 435/7.32, 436/805
• US Class Current: 435/7.32
• 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

Bioelectronic devices for the detection of estrogen include a collection of eukaryotic cells which harbor a recombinant lux gene from a high temperature microorganism wherein the gene is operably linked with a heterologous promoter gene. A detectable light-emitting lux gene product is expressed in the presence of the estrogen and detected by the device.

Claims

1. An apparatus for the detection of an estrogen or a xenoestrogen, comprising:

an integrated circuit chip, wherein said chip comprises an integrated light detection system, and

a biosensor comprising a collection of eukaryotic cells harboring a recombinant lux gene from a high temperature microorganism wherein said gene is operably linked with a heterologous promoter, wherein a detectable light-emitting lux gene product is expressed in the presence of said estrogen or xenoestrogen, light from said light-emitting lux gene product being detected by said light detection system.

2. The biosensor of claim 1 wherein the high temperature microorganism is a bioluminescent microorganism.

3. The biosensor of claim 2 wherein the bioluminescent microorganism is Xenorhabdus luminescens, Pseudomonas phosphoreum, or Photobacterium phosphoreum.

4. The biosensor of claim 1 wherein the estrogen or xenoestrogen is estrone, estradiol, estriol or an esterified estrogen.

5. A method for the detection of an estrogen or xenoestrogen compound comprising the steps of:

contacting a sample suspected of containing an estrogen or xenoestrogen with the apparatus of claim 1, and

detecting the presence of emitted light from a product expressed by the lux gene wherein said product expressed is induced in the presence of the estrogen or xenoestrogen compound.

6. The method of claim 5 wherein an intensity of the emitted light is directly related to a concentration of the estrogen or xenoestrogen compound.

7. A luminometer for the detection of an estrogen or xenoestrogen compound comprising the apparatus of claim 1, wherein said eukaryotic cell collection is held on the integrated chip surface and responds to the presence of estrogen or xenoestrogen by expressing a bioluminescent protein from the lux gene cassette, wherein bioluminescent protein is detected by the integrated light detection system.

8. The luminometer of claim 7 wherein the eukaryotic cell collection is encapsulated in a sol-gel matrix held on the integrated chip surface.

9. The luminometer of claim 8 wherein the sol-gel encapsulation matrix is selected from the group consisting of polydimethylsiloxane, polyvinyl alcohol/polyvinylpryidine copolymer, latex copolymer, agar/agarose, carrageenan, polyacrylamide, alginate, polyurethane/polycarbonyl sulfonate, polyvinyl alcohol and silicon glass.

10. The luminometer of claim 9 wherein the lux gene cassette is a luxCDABE gene.

11. A kit for detection of an estrogen or a xenoestrogen, said kit comprising:

at least one container containing at least one apparatus for the detection of an estrogen or a xenoestrogen according to claim 1, an instrument to place the biosensors upon the integrated light detection system, and instructions for use.