Compositions and methods for determining the presence of amphetamines in a sample suspected of containing amphetamine and/or methamphetamine

• US 5,328,828 A
• Filed: 07/20/1992
• Issued: 07/12/1994
• Est. Priority Date: 12/23/1988
• Status: Expired due to Term

• Current Assignee: Dade Behring Marburg GmbH (Siemens AG)
• Original Assignee: Syntec Incorporated (Atos SE)
• Inventors: Freeman, James V., Hsu, Chen-jung, Ullman, Edwin F., Lin, Cheng-I, Pirio, Marcel, Hu, Mae W.
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US07/916,568
• Time in Patent Office: 722 Days
• Field of Search: 435/7.9, 435/7.92, 436/536, 436/537, 436/538, 436/815, 436/816, 436/822, 436/548
• US Class Current: 435/7.9
• CPC Class Codes: C07C 233/18 having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
• CPC Class Codes: C07C 323/12 the carbon skeleton being acyclic and saturated
• CPC Class Codes: C07C 323/32 having at least one of the nitrogen atoms bound to an acyclic carbon atom of the carbon skeleton
• CPC Class Codes: C07C 323/41 Y being a hydrogen or an acyclic carbon atom
• CPC Class Codes: G01N 33/535 with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
• CPC Class Codes: G01N 33/946 CNS-stimulants, e.g. cocaine, amphetamines
• CPC Class Codes: Y10S 435/964 including enzyme-ligand conjugate production, e.g. reducing rate of nonproductive linkage
• CPC Class Codes: Y10S 436/816 Alkaloids, amphetamines, and barbiturates
• CPC Class Codes: Y10S 436/822 Identified hapten

Abstract

The instant invention is directed toward an immunoassay which can determine the presence of amphetamines in a sample suspected of containing amphetamine and/or methamphetamine by employing at least two conjugates, each comprised of a functionally similar label bound to an amphetamine analog and a methamphetamine analog respectively and an antibody to amphetamine and an antibody to methamphetamine wherein at least one of the antibodies is a monoclonal antibody.

Claims

1. A reagent system comprising (i) at least two conjugates, wherein in one conjugate an enzyme is bound to an amphetamine ligand analog and in the other conjugate said enzyme is bound to a methamphetamine ligand analog, (ii) an antibody to amphetamine, and (iii) an antibody to methamphetamine, wherein at least one of said antibodies is a monoclonal antibody and each of said antibodies is moderately cross-reactive with that conjugate which binds more strongly with the other antibody.

2. A reagent system according to claim 1 which further comprises any other necessary components of the signal producing system wherein said producing system is capable of producing a detectable signal wherein the amount of signal is related to the amount of amphetamines in the solution.

3. A reagent system according to claim 1 wherein both of said antibodies are monoclonal.

4. A method for the assay of amphetamines in a sample suspected of containing amphetamine and/or methamphetamine which comprises

A) combining in aqueous medium (a) said sample;

(b) two conjugates wherein in one conjugate an enzyme is conjugated to a ligand analogue of amphetamine and in the other conjugate said enzyme is conjugated to a ligand analogue of methamphetamine; and

(c) an antibody to amphetamine and an antibody to methamphetamine wherein at least one of said antibodies is a monoclonal antibody and each of said antibodies is moderately cross-reactive with that conjugate which binds more strongly with the other antibody under conditions allowing said antibodies to bind to said conjugates;

B) with or without separating the bound or unbound conjugates from the medium obtained from step A, contacting the conjugates obtained from step A with any necessary components of a signal producing system wherein a signal is produced that is related to the presence of amphetamines in said sample; and

C) determining the signal and correlating the signal to the presence of amphetamine and/or methamphetamine in the sample.

5. A method according to claim 4 wherein said enzyme is glucose-6-phosphate dehydrogenase.

6. A method according to claim 4 wherein neither bound nor unbound conjugates are separated from the medium in Step B.

7. A method according to claim 4 wherein either bound or unbound conjugates are separated from the medium in Step B.

8. A method according to claim 4 wherein both of said antibodies are monoclonal.

9. A kit for use in an assay for amphetamines in a sample suspected of containing amphetamine and/or methamphetamine which comprises the following components:

(a) two conjugates each comprised of an enzyme wherein in one conjugate enzyme is bound to an amphetamine ligand analogue and in the other conjugate enzyme is bound to a methamphetamine ligand analogue, and wherein said enzymes are derived from the same enzyme; and

(b) an antibody to amphetamine and an antibody to methamphetamine wherein at least one of said antibodies is a monoclonal antibody and each of said antibodies is moderately cross-reactive with that conjugate which binds more strongly with the other antibody.

10. A kit as defined in claim 9 which further comprises any other necessary members of the signal producing system.

11. A kit as defined in claim 9 wherein both of said antibodies are monoclonal.

12. A kit as defined in claim 9 wherein said enzyme is glucose-6-phosphate dehydrogenase.