Colors for chromogenic IHC and ISH staining with multi-dye quinone methide and tyramide conjugates

Patent metadata

Field	Value
Publication number	US-12287329-B2
Application number	US-202017081523-A
Country	US
Kind code	B2
Filing date	Oct 27, 2020
Priority date	Jun 28, 2016
Publication date	Apr 29, 2025
Grant date	Apr 29, 2025

Abstract

Disclosed herein are novel chromogenic conjugates, the conjugates comprising at least two detectable moieties.

First claim

The invention claimed is:

1. A two-chromogen conjugate including a terminal reactive group, comprising: (a) a first chromogen, and (b) a second chromogen, wherein the first and second chromogens are coupled via a lysine, and wherein the conjugate further comprises a single terminal reactive functional group coupled to the carboxyl group of the lysine, wherein the first chromogen is coupled to a first amine group of the lysine, wherein the second chromogen is coupled to the second amine group of the lysine, wherein the first and second chromogens are independently selected from the group consisting of a rhodamine or a derivative or an analog thereof, Dabsyl or a derivative or an analog thereof, Dabcyl or a derivative or an analog thereof, Cy3 or a derivative or an analog thereof, Cy7 or a derivative or an analog thereof, Cy3.5 or a derivative or an analog thereof, Cy3B or a derivative or an analog thereof, Cy5 or a derivative or an analog thereof, Cy5.5 or a derivative or an analog thereof, fluorescein or a derivative or an analog thereof, or TAMRA or a derivative or an analog thereof.

2. The two-chromogen conjugate of claim 1 , wherein the two-chromogen conjugate produces a blue or a blue-like color.

3. The two-chromogen conjugate of claim 1 , wherein the two-chromogen conjugate produces a red or a red-like color.

4. The two-chromogen conjugate of claim 1 , wherein one of the first and second chromogens comprises rhodamine or a derivative or an analog thereof, Dabsyl or a derivative or an analog thereof, or Dabcyl or a derivative or an analog thereof.

5. The two-chromogen conjugate of claim 1 , wherein the two-chromogen conjugate produces a green or a green-like color.

6. The two-chromogen conjugate of claim 1 , wherein the one of the first and second chromogens comprises rhodamine or a derivative or an analog thereof.

7. The two-chromogen conjugate of claim 6 , wherein another of the first and second chromogens comprises a cyanine-based-dye.

8. The two-chromogen conjugate of claim 7 , wherein the cyanine-based dye comprises Cy5 or a derivative or analog thereof.

9. The two-chromogen conjugate of claim 1 , wherein the two-conjugate chromogen consists of the first chromogen, the second chromogen, the linker, and the terminal reactive functional group.

10. The two-chromogen conjugate of claim 9 , wherein the first chromogen comprises rhodamine and wherein the second chromogen comprises a cyanine-based dye, and wherein the rhodamine is rhodamine 800.

11. A two-chromogen conjugate including a terminal reactive group, comprising: (a) a first chromogen, and (b) a second chromogen, wherein the first and second chromogens are coupled via a H 2 N—CH 2 —CH 2 —NH 2 linker, wherein the conjugate further comprises a single terminal reactive functional group, wherein the single terminal reactive group is included on one of the first or second chromogens, wherein the first and second chromogens are independently selected from the group consisting of (i) a rhodamine or a derivative or an analog thereof, (ii) Dabsyl or a derivative or an analog thereof, (iii) Dabcyl or a derivative or an analog thereof, (iv) Cy3 or a derivative or an analog thereof, (v) Cy7 or a derivative or an analog thereof, (vi) Cy3.5 or a derivative or an analog thereof, (vii) Cy3B or a derivative or an analog thereof, (viii) Cy5 or a derivative or an analog thereof, (ix) Cy5.5 or a derivative or an analog thereof; (x) TAMA or a derivative or an analog thereof, and (xi) Rhodamine 800 or a derivative or an analog thereof; and wherein the first and second chromogens are different; and where the first chromogen is coupled to a first amine group of the H 2 N—CH 2 —CH 2 —NH 2 linker and the second chromogen is coupled to the second amine group of the H 2 N—CH 2 —CH 2 —NH 2 linker.

Assignees

• Ventana Med Syst Inc

Inventors

• Ashworth-Sharpe Julia
• Kelly Brian D
• Lefever Mark
• Polaske Nathan W

Classifications

• C09B23/083

  five >CH- groups · CPC title

• C09B11/24 Primary

  Phthaleins containing amino groups {; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes} · CPC title

• G01N33/532

  Production of labelled immunochemicals · CPC title

• G01N33/583

  with non-fluorescent dye label · CPC title

• G01N33/582

  with fluorescent label · CPC title