Immunoassay for phenethylamines of the 2c and do sub-families

Patent metadata

Field	Value
Publication number	US-2018209998-A1
Application number	US-201815928595-A
Country	US
Kind code	A1
Filing date	Mar 22, 2018
Priority date	Aug 5, 2013
Publication date	Jul 26, 2018
Grant date	—

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

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:

2 . The antibody of claim 1 further characterised by being derived from an immunogen of structure:

3 . The antibody of claim 1 , wherein the antibody shows a sensitivity (IC 50 ) of less than 25 ng/mL for 2C-B.

4 . 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.

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

6 . The immunoassay method of claim 4 , 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.

7 . The immunoassay method of claim 6 , 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.

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

9 . The immunoassay method of claim 4 , wherein the sample is a blood, serum, plasma, urine or saliva sample.

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

11 . The immunoassay method of claim 4 , wherein the sample is from a human subject.

12 . 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.

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

14 . The substrate of claim 12 , wherein the substrate further comprises a second polyclonal a which 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 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 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.

Assignees

• Randox Laboratories Ltd

Inventors

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

Classifications

• C07K2317/33

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

• C07K16/44 Primary

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

• G01N33/94 Primary

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

• G01N33/946

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

• G01N2430/00

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