Polymer stabilization of chromogen solutions

Patent metadata

Field	Value
Publication number	US-9618429-B2
Application number	US-201313744504-A
Country	US
Kind code	B2
Filing date	Jan 18, 2013
Priority date	Jan 23, 2012
Publication date	Apr 11, 2017
Grant date	Apr 11, 2017

Abstract

Disclosed embodiments concern a composition comprising DAB chromogen, and/or derivative thereof, a stabilizer, and polymer capable of preventing or reducing DAB precipitation relative to a composition that does not comprise the polymer. Also disclosed herein is a method for using the disclosed composition and embodiments of a kit.

First claim

We claim:

1. A composition for chromogenic immunohistochemistry comprising in a solution:
a) a solvent,
b) a DAB chromogen or a derivative thereof,
c) a stabilizer, and
d) a polyethyleneimine polymer soluble in the solvent.

2. The composition of claim 1 , wherein the polyethyleneimine polymer is a 4 kDa HCl salt of polyethyleneimine.

3. The composition of claim 1 , wherein the polyethyleneimine polymer has a number average molecular weight of between about 1 kDa and about 500 kDa.

4. The composition of claim 1 , wherein the polyethyleneimine polymer is about 0.05 percent to about 10 percent of the solution by weight.

5. The composition of claim 1 , wherein the DAB chromogen or the derivative thereof has a concentration of about 0.1 mM to about 100 mM.

6. The composition of claim 1 , further comprising an enhancer selected from the group consisting of imidazole, 2-hydroxypyridine, and combinations thereof.

7. The composition of claim 1 , wherein the stabilizer is sodium metabisulfite.

8. The composition of claim 1 , wherein the polyethyleneimine polymer is configured to form a complex with the DAB chromogen or the derivative thereof so as to maintain solubility of the DAB chromogen or the derivative thereof in the solvent.

9. The composition of claim 1 , wherein the stabilizer and the polyethyleneimine polymer substantially prevent the DAB chromogen or the derivative thereof from decomposing and/or being oxidized under storage conditions.

10. A composition for chromogenic immunohistochemistry detection comprising a DAB chromogen, a stabilizer, and a polymer; wherein the stabilizer is sodium metabisulfite or sodium bisulfite, and the polymer is polyethyleneimine, wherein the composition is configured to remain stable under storage conditions so that the polymer complexes the DAB chromogen or the derivative thereof so as to maintain the amount of the DAB chromogen or the derivative thereof in the solution, and wherein the composition is configured so that the DAB precipitates under detection conditions to produce a signal suitable for chromogenic immunohistochemistry.

11. The composition of claim 10 , wherein the DAB chromogen is 3,3′-diaminobenzidine, and the stabilizer is sodium metabisulfite, and the polymer is polyethyleneimine.

12. The composition of claim 11 , wherein the composition comprises about 1 to about 15 mM 3,3′-diaminobenzidine, about 0.1 to about 6 mM sodium metabisulfite and about 0.05% to about 0.5% (w/w) polyethyleneimine.

Assignees

• Ventana Med Syst Inc

Inventors

• May Eric J
• Kelly Brian D
• Kosmeder Jerome W
• Morrison Larry
• Bieniarz Christopher

Classifications

• C12Q1/28 Primary

  involving peroxidase · CPC title

• G01N33/532

  Production of labelled immunochemicals · CPC title

• G01N1/30 Primary

  Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} · CPC title

• G01N33/535

  with enzyme label {or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates} · CPC title

• G01N33/581

  with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates) · CPC title