Haptens and immunoreactive agents and use thereof for producing family antibodies and immunoassays for quinolones

Patent metadata

Field	Value
Publication number	US-10150736-B2
Application number	US-201113988826-A
Country	US
Kind code	B2
Filing date	Nov 21, 2011
Priority date	Nov 23, 2010
Publication date	Dec 11, 2018
Grant date	Dec 11, 2018

Abstract

The invention relates to haptens, immunogens and secondary immunoreactive agents, to the use thereof for producing wide-spectrum antibodies against quinolone-type antibiotics, to the application thereof to immunochemical analysis techniques, and to a kit enabling the detection of said antibiotics in biological samples from food products of animal origin.

First claim

The invention claimed is:

1. A method for obtaining a polyclonal antibody against quinolone antibiotics comprising the following steps:
a) mixing an immunogenic carrier with a bifunctional linking compound in order to get an activated immunogenic carrier;
b) reacting the activated immunogenic carrier with a quinolone compound in order to get a conjugate, wherein said conjugate is 1-[3-(2-acetamide HCH)thiopropyI]-6-fluoro-7-(N-piperazinyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid, wherein HCH is horseshoe crab hemocyanin;
c) eliciting an in vivo humoral response against said conjugate obtained in (b) to generate the polyclonal antibody; and
d) isolating the polyclonal antibody generated in (c).

2. A method for detection and/or quantification of quinolone antibiotics, comprising the following steps:
obtaining a polyclonal antibody against the quinolone antibiotics according to the method of claim 1 ;
performing a competitive immunoassay, comprising:
(a) contacting said polyclonal antibody, a secondary immunoreagent, and a sample potentially containing the quinolone antibiotics, wherein the secondary immunoreagent is 7-[2-(BSA acetamide)ethylamino]-6-fluoro-4-oxo-1-propyl-1,4-dihydroquinoline-3-carboxylic acid or 7-[2(4-chloro-6-HRP-1,3,5,-triazine-2-ylamino)ethylamino]-6-fluoro-4-oxo-1-propyl-1,4-dihydroquinoline-3-carboxylic acid, wherein BSA is bovine serum albumin and HRP is horseradish peroxidase;
(b) executing a competition reaction between the secondary immunoreagent and the quinolone antibiotics in the sample for the polyclonal antibody;
(c) introducing components and/or reagents necessary to generate a measurable signal;

performing an analysis on the measurable signal for detecting and/or quantifying the presence of the quinolone antibiotics according to the results of the competition reaction.

3. The method according to claim 2 , wherein the sample is a product of animal origin intended for food consumption.

4. The method of claim 3 wherein the product of animal origin is milk.

Assignees

• Marco Colas Maria Pilar
• Sanchez Baeza Francisco Jose
• Gonzalez Pinacho Daniel
• Consejo Superior Investigacion

Inventors

• Marco Colas Maria Pilar
• Sánchez Baeza Francisco José
• González Pinacho Daniel

Classifications

• C07D215/233 Primary

  only one oxygen atom which is attached in position 4 · CPC title

• C07D401/12

  linked by a chain containing hetero atoms as chain links · CPC title

• G01N33/9446

  Antibacterials · CPC title

• C07D215/56 Primary

  with oxygen atoms in position 4 · CPC title

• C07K16/44

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