Method and kit for detecting, or determining, 3,4-methylenedioxymethamphetamine
View Patent ↗The invention provides a hapten, an immunogen comprising the hapten coupled to an antigenicity-conferring carrier material, a conjugate comprising the hapten coupled to a labelling agent, as well as, antibodies raised against the aforementioned immunogen and capable of binding with MDMA.
1. An immunogen of the following structural formula:
wherein a crosslinker comprises —R—X—, wherein R is a first bivalent group and X is a second bivalent group, and wherein W 1 is an antigenicity-conferring carrier material.
2. The immunogen of claim 1 , wherein R comprises:
—Z a —Y b —
wherein Z is selected from the group consisting of a substituted or unsubstituted, straight or branched chain, saturated or unsaturated alkylene moiety and a substituted or unsubstituted arylene moiety; Y is a substituted or unsubstituted, straight or branched chain, saturated or unsaturated alkylene moiety; a is 0 or 1 and b is 0 or 1, with the proviso that a and b cannot both be 0.
3. The immunogen of claim 2 , wherein Z is a substituted or unsubstituted phenylene moiety.
4. The immunogen of claim 2 , wherein b is 0, a is 1 and Z is a C 1-6 , substituted or unsubstituted, straight or branched chain, saturated or unsaturated alkylene moiety.
5. The immunogen of claim 4 , wherein Z is a C 3-4 unsubstituted, straight chain, saturated alkylene moiety.
6. The immunogen of claim 1 , wherein X is selected from the group consisting of a carboxylic acid or an ester thereof, an amine, a maleimide, a halocarboxylic acid or an ester thereof, a thiocarboxylic acid or an ester thereof, an aldehyde, a pyridyldithio and a vinylsulphone moiety.
7. The immunogen of claim 6 , wherein X is selected from the group consisting of a carboxylic acid (—COOH), an amine (—NH 2 ) and a thiocarboxylate ester (—S—CO—CH 3 ).
8. The immunogen of claim 1 , wherein the carrier material is selected from the group consisting of a protein, a protein fragment, a synthetic polypeptide and a semi-synthetic polypeptide.
9. Antibodies raised against the immunogen of claim 1 , the antibodies having specificity for MDMA and other methylenedioxy analogs.
10. The antibodies of claim 9 , wherein the antibodies bind a methylenedioxy structural epitope.
11. A process of preparing the antibodies according to claim 9 , the process comprising the steps of immunising an animal by repeated administration of the immunogen of claim 1 , and collecting the resulting serum from the immunised animal.
12. The process of claim 11 , the process comprising the steps of immunising a vertebrate animal by repeated administration of the immunogen of claim 1 , and collecting the resulting serum from the immunised animal.
13. The process of claim 11 , the process comprising the steps of immunising a mammalian animal by repeated administration of the immunogen of claim 1 , and collecting the resulting serum from the immunised animal.
14. A conjugate of the following structural formula:
wherein a crosslinker comprises —R—X—, wherein R is a first bivalent group and X is a second bivalent group, and wherein W 2 is a detectable labelling agent.
15. The conjugate of claim 14 , wherein R comprises:
—Z a —Y b —
wherein Z is selected from the group consisting of a substituted or unsubstituted, straight or branched chain, saturated or unsaturated alkylene moiety and a substituted or unsubstituted arylene moiety; Y is a substituted or unsubstituted, straight or branched chain, saturated or unsaturated alkylene moiety; a is 0 or 1 and b is 0 or 1, with the proviso that a and b cannot both be 0.
16. The conjugate of claim 15 , wherein Z comprises a substituted or unsubstituted phenylene moiety.
17. The conjugate of claim 15 , wherein b is 0, a is 1 and Z is a C 1-6 , substituted or unsubstituted, straight or branched chain, saturated or unsaturated alkylene moiety.
18. The conjugate of claim 17 , wherein Z is a C 3-4 unsubstituted, straight chain, saturated alkylene moiety.
19. The conjugate of claim 14 , wherein X is selected from the group consisting of a carboxylic acid or an ester thereof, an amine, a maleimide, a halocarboxylic acid or an ester thereof, a thiocarboxylic acid or an ester thereof, an aldehyde, a pyridyldithio and a vinylsulphone moiety.
20. The conjugate of claim 19 , wherein X is selected from the group consisting of a carboxylic acid (—COOH), an amine (—NH 2 ) and a thiocarboxylate ester (—S—CO—CH 3 ).
21. The conjugate of claim 14 , wherein the labelling agent is selected from the group consisting of an enzyme, a luminescent substance and a radioactive substance; or a mixture thereof.
22. A method for detecting or determining MDMA and its methylenedioxy analogs in a sample, the method comprising contacting the sample with at least one conjugate of claim 14 , and with at least one antibody of claim 9 ; detecting, or determining the quantity of, bound conjugate; and deducing from a calibration curve the presence of, or the amount of, MDMA and its methylenedioxy analogs in the sample.
23. A method for detecting or determining MDMA and its methylenedioxy analogs in a sample, the method comprising contacting the sample with at least one conjugate of claim 15 , and with at least one antibody of claim 10 ; detecting, or determining the quantity of, bound conjugate; and deducing from a calibration curve the presence of, or the amount of, MDMA and its methylenedioxy analogs in the sample.
24. A kit for detecting or determining MDMA and its methylenedioxy analogs, the kit including at least one conjugate of claim 14 , and at least one antibody of claim 9 .
25. A kit for detecting or determining MDMA and its methylenedioxy analogs, the kit including at least one conjugate of claim 15 , and at least one antibody of claim 10 .