Haptens, hapten conjugates, compositions thereof and method for their preparation and use

Patent metadata

Field	Value
Publication number	US-11092594-B2
Application number	US-201715665291-A
Country	US
Kind code	B2
Filing date	Jul 31, 2017
Priority date	Nov 1, 2006
Publication date	Aug 17, 2021
Grant date	Aug 17, 2021

Abstract

A method for performing a multiplexed diagnostic assay, such as for two or more different targets in a sample, is described. One embodiment comprised contacting the sample with two or more specific binding moieties that bind specifically to two or more different targets. The two or more specific binding moieties are conjugated to different haptens, and at least one of the haptens is an oxazole, a pyrazole, a thiazole, a nitroaryl compound other than dinitrophenyl, a benzofurazan, a triterpene, a urea, athiourea, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azo aryl, or a benzodiazepine. The sample is contacted with two or more different anti-hapten antibodies that can be detected separately. The two or more different anti-hapten antibodies may be conjugated to different detectable labels.

First claim

We claim:

1. A method of forming a first target-hapten complex in a sample including at least a first target, comprising:
contacting the sample with a first conjugate comprising (a) a first antibody that is specific to the first target, and (b) a first hapten coupled directly or indirectly to the first antibody, wherein the first hapten has a formula

2. The method of claim 1 , wherein the first hapten is derived from a compound having the formula:

3. The method of claim 1 , further comprising forming a second target hapten complex in the sample by contacting the sample with a second conjugate comprising (a) a second antibody that is specific to the second target, and (b) a second hapten coupled directly or indirectly to the second antibody; wherein the second hapten is selected from the group consisting of an oxazole, a pyrazole, a thiazole, a benzofurazan, a triterpene, a urea, a thiourea, a nitroaryl, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azoaryl and a benzodiazepine.

4. The method of claim 3 , wherein the second hapten is indirectly coupled to the second antibody through an alkylene oxide linker.

5. The method of claim 1 , further comprising forming a second target hapten complex in the sample by contacting the sample with a second conjugate comprising (a) a second antibody that is to a second in the sample, and (b) a second hapten, wherein the second hapten is a benzofurazan having a formula:

wherein one of R 5 , R 6 , R 7 and R 8 comprises a reactive group suitable for coupling to a linker such that the second hapten may be indirectly coupled to the second antibody; and wherein the other three of R 5 , R 6 , R 7 and R 8 are hydrogen.

6. The method of claim 1 , further comprising forming a second target hapten complex in the sample by contacting the sample with a second conjugate comprising (a) a second antibody that is specific to a second target present in the sample, and (b) a second hapten, wherein the second hapten is a benzofurazan having a formula:

wherein R 5 , R 6 , and R 8 are each hydrogen; and wherein R 7 is a reactive group suitable for coupling to a linker such that the second hapten may be indirectly coupled to the second antibody.

7. The method of claim 1 , further comprising forming a second target hapten complex in the sample by contacting the sample with a second conjugate comprising (a) a second antibody that is specific to a second target in the sample, and (b) a second hapten, wherein the second hapten is a nitropyrazole derived from a compound a formula:

8. A method of forming a first target-hapten complex in a sample including at least one target, comprising:
contacting the sample with a first antibody conjugated directly or indirectly to a first hapten, wherein the first antibody is specific to a first target within the sample; and
wherein the first hapten is derived from a heterobicyclic compound having a formula:

9. A method of forming a first target-hapten complex in a sample including at least one target, comprising:
contacting the sample with a first antibody conjugated directly or indirectly to a first hapten, wherein the first antibody is specific to a first target within the sample; and
wherein the first hapten has a formula

wherein R 1 and R 2 are independently selected from a hydroxyl group or a carboxyl group, and each Y is independently C, O, N, or S.

10. The method of claim 9 , wherein the first hapten has the formula:

11. The method of claim 9 , wherein the first hapten has the formula:

12. The method of claim 9 , wherein the first hapten is derived from a heterobicyclic compound having the formula:

13. The method of claim 9 , wherein at least one of the R 1 or R 2 group is bonded to a linker or directly to a carrier.

14. The method of claim 9 , further comprising forming a second target-hapten complex in the sample by contacting the sample with a second antibody that is conjugated to a second hapten; wherein the second antibody binds specifically to a second target present in the sample, and wherein the second hapten is a benzofurazan derived from a compound having a formula:

wherein one of R 5 , R 6 , R 7 and R 8 is a reactive group suitable for coupling to a linker; and wherein the other three of R 5 , R 6 , R 7 and R 8 are hydrogen.

15. The method of claim 9 , further comprising forming a second target hapten complex in the sample by contacting the sample with a second specific antibody that is conjugated to a second hapten; wherein the second antibody moiety binds specifically to a second target present in the sample, and wherein the second hapten is a benzofurazan derived from a compound having a formula:

wherein R 5 , R 6 , and R 8 are each hydrogen; and wherein R 7 is a reactive group suitable for coupling to a linker or a carrier molecule.

16. The method of claim 9 , further comprising forming a second target hapten complex in the sample by contacting the sample with a second specific antibody that is conjugated to a second hapten; wherein the second antibody binds specifically to a second target present in the sample, and wherein the second hapten is a nitropyrazole derived from a compound having a formula:

Assignees

• Ventana Med Syst Inc

Inventors

• Kosmeder Ii Jerome W
• Lefever Mark
• Johnson Donald
• Farrell Michael
• Zhilina Zhanna
• Bieniarz Christopher

Classifications

• C07D495/04 Primary

  Ortho-condensed systems · CPC title

• C07D277/76

  Sulfur atoms attached to a second hetero atom · CPC title

• C07D311/12

  substituted in position 3 and unsubstituted in position 7 · CPC title

• C07D271/08

  1,2,5-Oxadiazoles; Hydrogenated 1,2,5-oxadiazoles · CPC title

• C07D277/48

  by radicals derived from carbonic acid, or sulfur or nitrogen analogues thereof, e.g. carbonylguanidines · CPC title