# US 11,029,308 B2 · Methods for vapor detection and discrimination with mammalian odorant receptors expressed in heterologous cells

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- Number: US 11,029,308 B2
- Document ID: US11029308B2
- Status: Granted
- Kind: B2
- Application number: 16/345,878
- Filed: 2017-10-27
- Published: 2019-08-15
- Granted: 2021-06-08
- Earliest claimed: 2016-10-27
- Assignees: Duke University
- Applicants: Duke University
- Inventors: Hiro Matsunami
- Web page: https://patentgrep.com/patent/US11029308B2

## CPC

- G01N 33/5088: {of vertebrates}
- G01N 33/0001: {by organoleptic means}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present invention relates to an odorant receptor based odorant sensor system and related methods. In particular, systems and methods are provided permitting detection and discrimination of an odorant molecule in a vapor/gaseous phase using a panel of odorant receptors expressed in heterologous cells.

## Claims

### Claim 1

A method of identifying an odorant receptor ligand comprising the steps of:
a) providing a biological sample comprising cells expressing at least one odorant receptor and exposing the biological sample to at least one test compound in a vapor/gaseous phase,
wherein the one or more odorant receptors at least includes olfr145,
wherein the biological sample further expresses at least one of the following:
one or more proteins known to enhance cell surface localization of the odorant receptors selected from REEP1, RTP1 and RTP2,
one or more odorant receptor binding proteins selected from Lcn3, Lcn4, Lcn10, Lcn11, OBP1a, OBP1b, and OBP2b, and
one or more metabolic enzymes, wherein the one or more metabolic enzymes at least includes carboxyl esterase;
b) measuring a signal that is proportional to activation of one of the odorant receptors in said biological sample, and
c) comparing the measured level of activation of one of the odorant receptors determined in step b) to a reference activation level for the specific odorant receptor determined in the same conditions with a negative control where the biological sample has not been exposed to said at least one test compound in a vapor/gaseous phase.

### Claim 2 (depends on 1)

The method of claim 1,
wherein a level of activation determined in step b) in the biological sample after exposure to at least one test compound in a vapor/gaseous phase is higher than the level of activation determined in the same conditions with a negative control without exposure to the test compound in a vapor/gaseous phase indicates that said at least one test compound in a vapor/gaseous phase constitutes a ligand acting as an agonist for said at least one odorant receptor;
wherein a level of activation determined in step b) in the biological sample after exposure to at least one test compound in a vapor/gaseous phase is lower than the level of activation determined in the same conditions with a negative control without exposure to the test compound in a vapor/gaseous phase indicates that said at least one test compound in a vapor/gaseous phase constitutes a ligand acting as an antagonist for said at least one odorant receptor.

### Claim 3 (depends on 1)

The method of claim 1, wherein said biological sample comprises a tissue from the olfactory system; and/or wherein the biological sample comprises heterologous cells.

### Claim 4 (depends on 1)

The method of claim 1, wherein a luminometer is used for the step of measuring a signal that is proportional to activation of one of the odorant receptors in said biological sample.

### Claim 5

A method for identifying an odorant receptor ligand, comprising:
a) providing
i) providing a biological sample comprising cells expressing at least one odorant receptor and exposing the biological sample to at least one test compound in a vapor/gaseous phase,
wherein the one or more odorant receptors at least includes olfr145,
wherein the biological sample further expresses at least one of the following:
one or more proteins known to enhance cell surface localization of the odorant receptors selected from REEP1, RTP1 and RTP2,
one or more odorant receptor binding proteins selected from Lcn3, Lcn4, Lcn10, Lcn11, OBP1a, OBP1b, and OBP2b, and
one or more metabolic enzymes, wherein the one or more metabolic enzymes at least includes carboxyl esterase, and
ii) a test compound in a vapor/gaseous phase, wherein said test compound in a vapor/gaseous phase is an odiferous molecule;
b) exposing said test compound in a vapor/gaseous phase to said biological sample in an in vitro setting; and
c) detecting the activity of said odorant receptor, wherein said detecting comprises detecting a reporting agent.

### Claim 6 (depends on 5)

The method of claim 5, wherein one or more of said odorant receptors is a human odorant receptor, a murine odorant receptor, or a synthetic odorant receptor.

### Claim 7 (depends on 5)

The method of claim 5, further comprising the step of d) detecting the presence or absence of an odorant receptor ligand based upon said activity.