U-47700 immunoassay

Patent metadata

Field	Value
Publication number	US-10815192-B2
Application number	US-201815903361-A
Country	US
Kind code	B2
Filing date	Feb 23, 2018
Priority date	Feb 23, 2017
Publication date	Oct 27, 2020
Grant date	Oct 27, 2020

Abstract

Antibodies, immunoassay methods and kits for the detection and determination of 3,4,-dichloro-N-[2-(dimethylamino)cyclohexyl]-N-methylbenzamide and 3,4,-dichloro-N-[2-(methylamino)cyclohexyl]-N-methylbenzamide, as well as the precursory immunogens, are described.

First claim

The invention claimed is:

1. A polyclonal antibody having a cross-reactivity to 3,4-dichloro-N-[2-(dimethylamino)cyclohexyl]-N-methylbenzamide (U-47700) of 100.00%, <<3.610% for AH-7921, 20% for desmethyl U-47700, <<3.610% for MT-45, <<3.610% for nor AH-7921 and <<3.610% for U-50488, wherein the polyclonal antibody is raised to an immunogen comprising a conjugate of N-desmethyl U-47700 N-propionic acid with Keyhole Limpet Hemocyanin (KLH); and
wherein said cross-reactivity is measured in the presence of a conjugate of N-desmethyl U-47700 acetic acid homocysteinethiolactone with horseradish peroxidase (HRP) as a tracer.

2. The polyclonal antibody of claim 1 , wherein the polyclonal antibody has an IC50 of 0.722 ng/ml for U-47700, >>20.00 ng/ml for AH-7921, 3.61 ng/ml for desmethyl U-47700, >>20.00 ng/ml for MT-45, >>20.00 ng/ml for nor AH-7921 and >>20.00 ng/ml for U-50488.

3. An immunoassay method of detecting or determining 3,4-dichloro-N-[2-(dimethylamino)cyclohexyl]-N-methylbenzamide and 3,4-dichloro-N-[2-(methylamino) cyclohexyl]-N-methylbenzamide in a solution or an in vitro sample taken from an individual with one or more detecting agents and the polyclonal antibody of claim 1 ; measuring the signal or signals produced by the one or more detecting agents; and deducing the presence of, or amount of, 3,4-dichloro-N-[2-(dimethylamino)cyclohexyl]-N-methyl benzamide and 3,4-dichloro-N-[2-(methylamino)cyclohexyl]-N-methylbenzamide.

4. A substrate comprising the polyclonal antibody of claim 1 .

5. The substrate of claim 4 , wherein the substrate is a microtitre plate or a biochip.

6. The substrate of claim 5 , wherein the biochip is a ceramic biochip.

7. A kit comprising the polyclonal antibody of claim 1 .

8. A method of making an antibody comprising:
administering an immunogen comprising a conjugate of N-desmethyl U-47700 N-propionic acid with Keyhole Limpet Hemocyanin (KLH) to a mammal; and purifying the resultant antibody.

9. The method of claim 8 , wherein the antibody is polyclonal.

Assignees

• Randox Laboratories Ltd

Inventors

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

Classifications

• C07K16/44 Primary

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

• C07K16/16

  against material from plants · CPC title

• C07K2317/33

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

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title

• C07K16/18

  against material from animals or humans · CPC title