Multiplexed Olfactory Receptor-Based Microsurface Plasmon Polariton Detector

• US 20120021932A1
• Filed: 01/28/2011
• Published: 01/26/2012
• Est. Priority Date: 07/31/2008
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Mershin, Andreas, Miura, Yoshikatsu, Zhang, Shuguang, Cook, Brian, Kaiser, Liselotte, Bikker, Johanna F., Niwa, Daisuke, Ohnishi, Dai, Tazuke, Atsushi
• Granted Patent: US 8,748,111 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07K 2/00 Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 21/554 detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
• CPC Class Codes: G01N 2201/06113 Coherent sources; lasers
• CPC Class Codes: G01N 2201/08 Optical fibres; light guides
• CPC Class Codes: G01N 33/0001 by organoleptic means
• CPC Class Codes: G01N 33/0031 comprising two or more sensors, e.g. a sensor array
• CPC Class Codes: G01N 33/02 Food
• CPC Class Codes: G01N 33/497 of gaseous biological material, e.g. breath
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

The invention provides a bio-sensing nanodevice comprising: a stabilized G-protein coupled receptor on a support, a real time receptor-ligand binding detection method, a test composition delivery system and a test composition recognition program. The G-protein coupled receptor can be stabilized using surfactant peptide. The nanodevice provides a greater surface area for better precision and sensitivity to odorant detection. The invention further provides a microfluidic chip containing a stabilized G-protein coupled receptor immobilized on a support, and arranged in at least two dimensional microarray system. The invention also provides a method of delivering odorant comprising the step of manipulating the bubbles in complex microfluidic networks wherein the bubbles travel in a microfluidic channel carrying a variety of gas samples to a precise location on a chip. The invention further provides method of fabricating hOR17-4 olfactory receptor.

Claims

1. A bio-sensing nanodevice comprising:

a stabilized G-protein coupled receptor [2] on a support comprising a receptor binding surface, a real time receptor-ligand binding detection method [6], a test composition delivery system [5] and a test composition recognition program [7] wherein the support comprises a dielectric/metal/dielectric sandwich structure.

2. The bio-sensing device of claim 1, wherein a laser light source is irradiated across the metal layer on a horizontal plane causing Plasmon to be generated on both the top and bottom of the metal layer.

3. The bio-sensing device of claim 2, wherein the laser light source is irradiated to the olfactory receptor via light fiber.

4. The bio-sensing nanodevice of claim 1, wherein the stabilized G-protein coupled receptor [2] is stabilized with surfactant peptides or with a detergent selected from the group consisting of n-Tetradecylphosphocholine 12, 13, 14, 15, 16 (also called FC12, FC13, FC14, FC15, FC16), digitonin and trimix [CHAPS (3-cholamidopropyl-dimethylammonio-1-propane sulfonate), CHS (Cholesteryl hemisuccinate), and DDM [n-dodecyl-beta-D-maltopyranoside)].

5. The bio-sensing nanodevice of claim 4, wherein the surfactant peptides have a formula selected from the group consisting of:

a. (Φ

)_m(+)_n 

 

(Formula (1)),

b. (+)_n(Φ

)_m 

 

(Formula (2)),

c. (Φ

)_m(−

)_n 

 

(Formula (3)),

d. (−

)_n(Φ

)_m 

 

(Formula (4)),

e. (−

)_n(Φ

)_m(−

)_n 

 

(Formula (5)),

f. (+)_n(Φ

)_m(+)_n 

 

(Formula (6)),

g. (Φ

)_m(−

)_n(Φ

)_m 

 

(Formula (7)),

h. (Φ

)_m(+)_n(Φ

)_m 

 

(Formula (8)),

i. (+)_n(Φ

)_m(−

)_n 

 

(Formula (9)),

j. (−

)_n(Φ

)_m(+)_n 

 

(Formula (10)),

k. (τ

)_m(Φ

)_m 

 

(Formula (11)),

l. (Φ

)_m(τ

)_n 

 

(Formula (12)),

m. (τ

)_n(Φ

)_m(τ

)_n, 

 

(Formula (13)), and

n. (Φ

)_m(τ

)_n(Φ

)_m, 

 

(Formula (14)),

wherein;

(Φ

) represents independently for each occurrence a natural or non-natural amino acid comprising a hydrophobic sidechain;

preferably alanine, valine, leucine, isoleucine or proline;

(+) represents independently for each occurrence a natural or non-natural amino acid comprising a sidechain that is cationic at physiological pH;

preferably histidine, lysine or arginine;

(−

) represents independently for each occurrence a natural or non-natural amino acid comprising a sidechain that is anionic at physiological pH;

preferably aspartic acid or glutamic acid;

(τ

) represents independently a polar amino acid containing a non-charged side chain at physiological pH, for example, serine, threonine, asparagine and glutamine;

wherein the terminal amino acids are optionally substituted;

m for each occurrence represents an integer greater than or equal to 5; and

n for each occurrence represents an integer greater than or equal to 1.

6. The bio-sensing nanodevice of claim 5, wherein the surfactant peptide is selected from the group consisting of:

7. The bio-sensing nanodevice of claim 5, wherein the surfactant peptide is AAAAAAD (SEQ ID NO:

1), VVVVVVD (SEQ ID NO;

     2) or AAAAAAK (SEQ ID NO;

     47).

8. The bio-sensing nanodevice of claim 1, wherein the G-protein coupled receptor [2] is an olfactory receptor and the test composition is an odorant.

9. The bio-sensing nanodevice of claim 8, wherein the olfactory receptors is a mammalian receptor.

10. The bio-sensing nanodevice of claim 8, wherein the olfactory receptor is hOR17-4.

11. The bio-sensing nanodevice of claim 1, wherein the support [15] comprises a material selected from gold, silver, platinum, glass, silicon, silica, polystyrene and polymer films, which is coated or non-coated.

12. The bio-sensing nanodevice of claim 1, wherein the receptor-ligand binding detection method [6] is selected from Surface Plasmon Resonance (SPR), micro-slot nuclear magnetic resonance (microNMR), scanning tunneling microscopy (STM)/impedance spectroscopy, ZnO nanowire surface passivation, optical microscope(s), confocal microscope(s), and reading device(s) using a laser light source.

13. The bio-sensing nanodevice of claim 12, wherein the optical microscope is selected from ultraviolet-visible absorption and fluorescence resonant energy transfer.

14. The bio-sensing nanodevice of claim 1, wherein the test composition delivery system [5] is passive exposure to air or microfluidic bubble logic operation.

15. The bio-sensing nanodevice of claim 14, wherein the microfluidic bubble logic operation comprises micron-sized droplets and bubbles of chemical in a microfluidic chip [11].

16. The bio-sensing nanodevice of claim 1, wherein the test composition recognition program [7] comprises a 1 dimensional peak-recognition program.

24. The microfluidic chip [11] of claim 17, comprising a plurality of sensors each including a nanodevice in accordance with any one of claims 1 through 15, wherein the number of sensors is between about 36 and about 900.

25. The microfluidic chip [11] of claim 24, wherein the microfluidic chip contains a plurality of receptors and each sensor contains one receptor and is connected by a microfluidic channel configured to permit a bubble containing a test compound to be delivered.

26. The microfluidic chip [11] of claim 25, wherein the sensors are connected serially, in parallel or a combination thereof.

17. A microfluidic chip [11] for use in detecting a test composition comprises at least one stabilized G-protein coupled receptor [2] immobilized on a support comprising a receptor binding surface, and arranged in at least two dimensional microarray system wherein the support comprises a dielectric/metal/dielectric sandwich structure.

18. The microfluidic chip [11] of claim 17, wherein a laser light source is irradiated across the metal layer on a horizontal plane causing Plasmon to be generated on both the top and bottom of the metal layer.

19. The microfluidic [11] chip of claim 18, wherein the laser light source is irradiated to the olfactory receptor via light fiber.

20. The microfluidic chip [11] of claim 17, wherein the stabilized G-protein coupled receptors [2] are stabilized with surfactant peptides.

21. The microfluidic chip [11] of claim 17, wherein the support comprises a material selected from gold, silver, glass, silicon, silica, polystyrene and polymer films, which is coated or non-coated.

22. The microfluidic chip [11] of claim 17, wherein the G-protein coupled receptor [2] is an olfactory receptor.

23. The microfluidic chip [11] of claim 17, wherein the olfactory receptor is hOR17-4.

27. The microfluidic chip of claim 17, wherein the microfluidic chip is disposable or non-disposable.

28. A method of delivering odorant comprising the step of manipulating the bubbles in complex microfluidic networks wherein the bubbles travel in a microfluidic channel carrying a variety of gas samples to a precise location on a chip, using a microfluidic chip [11] in accordance with claim 17.

29. A method of identifying an odor from a gas sample comprising the steps of:

(a) contacting a bubble containing the gas sample to a microfluidic chip [11] in accordance with claim 17;

(b) identifying the receptors that are and/or are not activated by the gas sample; and

(c) comparing the receptors identified in step (b) with a standard.

30. The method of claim 29, wherein the standard is selected from the group consisting of a chemical compound, biological substances (including bacteria, mold, mildew, fungi, and viruses), illegal substances, controlled substances, pathogens, toxins, contaminants, foods, perfumes, and human odors (including breath, body odors indicative of a disease or condition).