Immunoassay for phenethylamines of the 2C and DO sub-families

Patent metadata

Field	Value
Publication number	US-10775394-B2
Application number	US-201815928595-A
Country	US
Kind code	B2
Filing date	Mar 22, 2018
Priority date	Aug 5, 2013
Publication date	Sep 15, 2020
Grant date	Sep 15, 2020

Abstract

Immunoassay methods and their requisite components for the detection and determination of phenethylamines of the 2C and DO sub-families are described.

First claim

The invention claimed is:

1. A polyclonal antibody which specifically binds to 1-(4-bromo-2,5-dimethoxyphenyl)ethan-2-amine (2C-B), 1-(2,5-dimethoxy-4-propylphenyl)ethan-2-amine (2C-P), 1-(2,5-dimethoxy-4-ethylphenyl)ethan-2-amine (2C-E), 1-(2,5-dimethoxy-4-ethylthiophenyl)ethan-2-amine (2C-T2), 1-(4-chloro-2,5-dimethoxyphenyl)ethan-2-amine (2C-C), 1-(2,5-dimethoxy-4-propylthiophenyl)ethan-2-amine (2C-T-7), 1-(2,5-dimethoxy4-methylphenyl)ethan-2-amine (2C-D), and 1-(4-cyano-2,5-dimethoxyphenyl)ethan-2-amine (2C-CN), wherein the polyclonal antibody shows a cross-reactivity of less than 1.0% for each of 1-(4-bromo-2,5-dimethoxyphenyl)propan-2-amine (DOB), (+)-Phenethylamine, Dimethylamylamine, when compared to 100% cross-reactivity for 2C-B, wherein said cross reactivity is measured in the presence of a conjugate of structure:

wherein the polyclonal antibody is derived from an immunogen of structure:

2. The polyclonal antibody of claim 1 , wherein the polyclonal antibody shows a sensitivity (IC 50 ) of 24.618 ng/mL for 2C-B.

3. An immunoassay method of detecting phenethylamines of a 2C sub-family, the method comprising contacting a sample, with the polyclonal antibody of claim 1 and one or more detecting agents, and measuring a signal or signals produced by the one or more detecting agents; and deducing from a calibration curve the presence of, or amount of, phenethylamines of the 2C sub-family.

4. The immunoassay method of claim 3 , wherein the one or more detecting agents are selected from the group consisting of an enzyme, a substance having fluorescent properties and a substance having a radioactive label.

5. The immunoassay method of claim 3 , wherein the immunoassay method comprises a competitive assay format in which phenethylamines of the 2C sub-family compete with the one or more detecting agents in binding with the binding sites of the polyclonal antibody, wherein the more phenethylamines of the 2C sub-family present in the sample, the less detecting agent that binds to the polyclonal antibody and the lower the measured signal.

6. The immunoassay method of claim 5 , wherein the one or more detecting agents comprise a detecting agent that has a structure that is similar to phenethylamines of the 2C sub-family in which an enzyme or a substance having fluorescent properties has been conjugated to the structure of the detecting agent.

7. The immunoassay method of claim 6 , wherein the detecting agent has a structure of:

8. The immunoassay method of claim 3 , wherein the sample is a blood, serum, plasma, urine or saliva sample.

9. The immunoassay method of claim 8 , wherein the sample is a serum, plasma or urine sample.

10. The immunoassay method of claim 3 , wherein the sample is from a human subject.

11. A substrate which comprises the polyclonal antibody of claim 1 , wherein the polyclonal antibody is either absorbed to the substrate or attached to the substrate via covalent bonds.

12. The substrate of claim 11 , wherein the substrate is a biochip.

13. The substrate of claim 11 , wherein the substrate further comprises a second polyclonal antibody, wherein the second polyclonal antibody specifically binds to 1-(4-bromo-2,5-dimethoxyphenyl)propan-2-amine (DOB), 1-(2,5-dimethoxy-4-methylphenyl)propan-2-amine (DOM), and 1-(2,5-dimethoxy-4-nitrophenyl)propan-2-amine (DON), wherein the second polyclonal antibody shows a cross-reactivity of less than 1.0% for each of Dimethylamylamine, Methamphetamine, Amphetamine, (+)-Phenethylamine, 1-(4-bromo-2,5-dimethoxyphenyl)ethan-2-amine (2C-B), 1-(2,5-dimethoxy-4-propylphenyl)ethan-2-amine (2C-P), 1-(2,5-dimethoxy-4-ethylphenyl)ethan-2-amine (2C-E), 1-(2,5-dimethoxy-4-ethylthiophenyl)ethan-2-amine (2C-T2), 1-(4-chloro-2,5-dimethoxyphenyl)ethan-2-amine (2C-C), 1-(2,5-dimethoxy-4-propylthiophenyl)ethan-2-amine (2C-T-7), 1-(2,5-dimethoxy4-methylphenyl)ethan-2-amine (2C-D), 1-(4-cyano-2,5-dimethoxyphenyl)ethan-2-amine (2C-CN), when compared to 100% cross-reactivity for DOB, wherein said second polyclonal antibody cross reactivity is measured in the presence of a conjugate of structure

wherein the second polyclonal antibody is either absorbed to the substrate or attached to the substrate via covalent bonds, and wherein the second polyclonal antibody is derived from an immunogen of structure:

Assignees

• Randox Laboratories Ltd

Inventors

• Benchikh Elouard
• Fitzgerald Peter
• Mcconnell Ivan
• Lowry Philip

Classifications

• C07K16/44 Primary

  against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} · CPC title

• G01N33/946

  CNS-stimulants, e.g. cocaine, amphetamines · CPC title

• C07K2317/33

  Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity · CPC title

• G01N33/94 Primary

  involving narcotics {or drugs or pharmaceuticals, neurotransmitters or associated receptors} · CPC title

• G01N2430/00

  Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes · CPC title