US2014364330A1PendingUtilityA1
Multiplexed Olfactory Receptor-Based Microsurface Plasmon Polariton Detector
Assignee: MASSACHUSETTS INST TECHNOLOGYPriority: Jul 31, 2008Filed: Jun 9, 2014Published: Dec 11, 2014
Est. expiryJul 31, 2028(~2.1 yrs left)· nominal 20-yr term from priority
Inventors:Andreas MershinBrian CookLiselotte KaiserJohanna F. BikkerYoshikatsu MiuraDaisuke NiwaDai OhnishiAtsushi TazukeShuguang Zhang
G01N 2201/06113G01N 2201/08G01N 21/553G01N 33/0031G01N 21/554B82Y 30/00C07K 2/00G01N 33/02G01N 33/497G01N 33/0001G01N 33/54373G01N 33/54393G01N 33/566
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Claims
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
exact text as granted — not AI-modified1 . 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) n (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:
(SEQ ID NO: 1)
AAAAAAD,
(SEQ ID NO: 2)
VVVVVVD,
(SEQ ID NO: 3)
AAAAAADD,
(SEQ ID NO: 4)
VVVVVVDD,
(SEQ ID NO: 5)
LLLLLLDD,
(SEQ ID NO: 6)
KKIIIIII,
(SEQ ID NO: 7)
KKLLLLLL,
(SEQ ID NO: 8)
KKAAAAAAA,
(SEQ ID NO: 9)
KKVVVVVV,
(SEQ ID NO: 10)
DDDDDDDDDDAAAAAAAAAA,
(SEQ ID NO: 11)
AAAAAAAAAADDDDDDDDDD,
(SEQ ID NO: 12)
EEEEEEEEEEAAAAAAAAAA,
(SEQ ID NO: 13)
AAAAAAAAAAEEEEEEEEEE,
(SEQ ID NO: 14)
DDDDDDDDDDVVVVVVVVVV,
(SEQ ID NO: 15)
VVVVVVVVVVDDDDDDDDDD,
(SEQ ID NO: 16)
DDDDDDDDDDPPPPPPPPPP,
(SEQ ID NO: 17)
PPPPPPPPPPDDDDDDDDDD,
(SEQ ID NO: 18)
AAAAAAAAAAHHHHHHHHHH,
(SEQ ID NO: 19)
HHHHHHHHHHAAAAAAAAAA,
(SEQ ID NO: 20)
KKKKKKKKKKAAAAAAAAAA,
(SEQ ID NO: 21)
AAAAAAAAAAKKKKKKKKKK,
(SEQ ID NO: 22)
RRRRRRRRRRAAAAAAAAAA,
(SEQ ID NO: 23)
AAAAAAAAAARRRRRRRRRR,
(SEQ ID NO: 24)
DDDDDDDDDDAAAAAAAAAADDDDDDDDDD,
(SEQ ID NO: 25)
EEEEEEEEEEAAAAAAAAAAEEEEEEEEEE,
(SEQ ID NO: 26)
DDDDDDDDDDVVVVVVVVVVDDDDDDDDDD,
(SEQ ID NO: 27)
DDDDDDDDDDPPPPPPPPPPDDDDDDDDDD,
(SEQ ID NO: 28)
HHHHHHHHHHAAAAAAAAAAHHHHHHHHHH,
(SEQ ID NO: 29)
KKKKKKKKKKAAAAAAAAAAKKKKKKKKKK,
(SEQ ID NO: 30)
RRRRRRRRRRAAAAAAAAAARRRRRRRRRR,
(SEQ ID NO: 31)
AAAAAAAAAADDDDDDDDDDAAAAAAAAAA,
(SEQ ID NO: 32)
AAAAAAAAAAEEEEEEEEEEAAAAAAAAAA,
(SEQ ID NO: 33)
VVVVVVVVVVDDDDDDDDDDVVVVVVVVVV,
(SEQ ID NO: 34)
PPPPPPPPPPDDDDDDDDDDPPPPPPPPPP,
(SEQ ID NO: 35)
AAAAAAAAAAHHHHHHHHHHAAAAAAAAAA,
(SEQ ID NO: 36)
AAAAAAAAAAKKKKKKKKKKAAAAAAAAAA,
(SEQ ID NO: 37)
AAAAAAAAAARRRRRRRRRRAAAAAAAAAA,
(SEQ ID NO: 38)
KKKKKKKKKKAAAAAAAAAADDDDDDDDDD,
(SEQ ID NO: 39)
KKKKKKKKKKAAAAAAAAAAEEEEEEEEEE,
(SEQ ID NO: 40)
RRRRRRRRRRVVVVVVVVVVDDDDDDDDDD,
(SEQ ID NO: 41)
KKKKKKKKKKPPPPPPPPPPDDDDDDDDDD,
(SEQ ID NO: 42)
HHHHHHHHHHAAAAAAAAAAEEEEEEEEEE,
and
(SEQ ID NO: 47)
AAAAAAK.
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.
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.
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.
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).Cited by (0)
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