Haptens, immunogens, antibodies and conjugates to 2-oxo-3-hydroxy-LSD
View Patent ↗The invention provides a hapten derivatized with a crosslinker at the nitrogen of the 8β-carboxamide of 2-oxo-3-hydroxy LSD. The invention also provides an immunogen comprising the aforementioned hapten coupled to an antigenicity-conferring carrier material; a conjugate comprising the aforementioned hapten coupled to a labelling agent, as well as, antibodies raised against the aforementioned immunogen and capable of binding with at least the 3-hydroxy-2-pyrrolidone structural epitope of 2-oxo-3-hydroxy LSD.
1. A hapten having the following structural formula:
wherein R is a bivalent link and X is a functional group for coupling to an antigenicity-conferring carrier material or to a detectable labeling agent.
2. The hapten of claim 1 , wherein R is selected from the group consisting of an alkylene moiety; a cycloalkylene moiety; and an arylene moiety; and X is independently selected from the group consisting of a carboxylic acid or an ester thereof, an aldehyde, a thiocarboxylic acid or an ester thereof, and a halocarboxylic acid or an ester thereof.
3. The hapten of claim 2 , wherein X is independently selected from thioacetyl and haloacetyl.
4. The hapten of claim 2 , wherein R is a C 1-6 straight chain, saturated alkylene moiety.
5. The hapten of claim 4 , wherein R is a C 2-3 straight chain, saturated alkylene moiety.
6. The hapten of claim 2 , wherein X is carboxylic acid.
7. The hapten of claim 1 , the hapten being the following hapten derivative of 2-oxo-3-hydroxy-LSD:
8. The hapten of claim 5 , wherein R is a saturated, unsubstituted straight chain alkylene group having 2 carbon atoms.
9. An immunogen having the following structural formula:
wherein R is a first bivalent group, X 1 is a second bivalent group and G is an antigenicity-conferring carrier material.
10. The immunogen of claim 9 , wherein R is selected from the group consisting of an alkylene moiety; a cycloalkylene moiety; and an arylene moiety; and X 1 is independently selected from the group consisting of a carbonyl (CO), a methylene (CH 2 ), a sulphur atom (S) and a COCH 2 group.
11. The immunogen of claim 9 , having the following structural formula:
12. An antibody raised against the immunogen of claim 9 , wherein the antibody binds at least the 3-hydroxy-2-pyrrolidone structural epitope of 2-oxo-3-hydroxy-LSD and wherein the antibody has a cross-reactivity of less than 25% for LSD and nor LSD, when compared to 100% for 2-oxo-3-hydroxy-LSD.
13. An antibody raised against the immunogen of claim 10 , wherein the antibody binds at least the 3-hydroxy-2-pyrrolidone structural epitope of 2-oxo-3-hydroxy-LSD and wherein the antibody has a cross-reactivity of less than 25% for LSD and nor LSD, when compared to 100% for 2-oxo-3-hydroxy-LSD.
14. An antibody raised against the immunogen of claim 11 , wherein the antibody binds at least the 3-hydroxy-2-pyrrolidone structural epitope of 2-oxo-3-hydroxy-LSD and wherein the antibody has a cross-reactivity of less than 25% for LSD and nor LSD, when compared to 100% for 2-oxo-3-hydroxy-LSD.
15. A conjugate having the following structural formula:
wherein R is a first bivalent group, X 1 is a second bivalent group and H that is attached to X 1 is a detectable labeling agent.
16. The conjugate of claim 15 , wherein R is selected from the group consisting of an alkylene moiety; a cycloalkylene moiety; and an arylene moiety; and X 1 is independently selected from the group consisting of a carbonyl (CO), a methylene (CH 2 ) a sulphur atom (S) and a COCH 2 group.
17. The conjugate of claim 15 , having the following structural formula:
18. The conjugate of claim 15 , wherein the labelling agent is selected from the group consisting of an enzyme, a luminescent substance and a radioactive substance.
19. A process of preparing the antibody of claim 12 , the process comprising the steps of immunising an animal by repeated administration of the immunogen of claim 9 , and collecting the resulting serum from the immunised animal.
20. A process of preparing the antibody of claim 13 , the process comprising the steps of immunising an animal by repeated administration of the immunogen of claim 10 , and collecting the resulting serum from the immunised animal.
21. A process of preparing the antibody of claim 14 , the process comprising the steps of imrnunising an animal by repeated administration of the immunogen of claim 11 , and collecting the resulting serum from the immunised animal.
22. A method for detecting or determining 2-oxo-3-hydroxy-LSD in a sample, the method comprising contacting the sample with the conjugate of claim 15 and with at least one antibody of claim 12 ; detecting or determining bound conjugate; and deducing from a calibration curve the presence of, or the amount of, 2-oxo-3-hydroxy-LSD in the sample.
23. A method for detecting or determining 2-oxo-3-hydroxy-LSD in a sample, the method comprising contacting the sample with at least one conjugate of claim 16 and with at least one antibody of claim 13 ; detecting or determining bound conjugate; and deducing from a calibration curve the presence of, or the amount of, 2-oxo-3-hydroxy-LSD in the sample.
24. A kit for detecting or determining 2-oxo-3-hydroxy-LSD, the kit comprising at least one conjugate of claim 15 ; and at least one antibody of claim 12 .
25. A kit for detecting or determining 2-oxo-3-hydroxy-LSD, the kit comprising at least one conjugate of claim 16 ; and at least one antibody of claim 13 .
26. An antibody raised against an immunogen of claim 9 having specificity for 2-oxo-3-hydroxy LSD, characterized by having cross reactivity of less than 5% for both LSD and nor LSD, when compared to 100% for 2-oxo-3-hydroxy-LSD.
27. An antibody raised against an immunogen of claim 10 having specificity for 2-oxo-3-hydroxy LSD, characterized by having cross reactivity of less than 5% for both LSD and nor LSD, when compared to 100% for 2-oxo-3-hydroxy-LSD.
28. An antibody raised against an immunogen of claim 11 having specificity for 2-oxo-3-hydroxy LSD, charaterized by having cross reactivity of les than 5% for both LSD and nor LSD, when compared to 100% for 2-oxo-3-hydroxy-LSD.
29. A method for detecting or determining 2-oxo-3-hydroxy-LSD in a sample, the method comprising contacting the sample with conjugate of claim 17 , and with at least one antibody of claim 14 ; detecting or determining bound conjugate; and deducing from a calibration curve the presence of, or the amount of, 2-oxo-3hydroxy-LSD in the sample.
30. A kit for detecting or determining 2-oxo-3-hydroxy-LSD, the kit comprising the conjugate of claim 17 ; and at least one antibody of claim 14 .