Chemiluminescent and fluorescent substrate pads and uses thereof

Patent metadata

Field	Value
Publication number	US-10722882-B2
Application number	US-201715667864-A
Country	US
Kind code	B2
Filing date	Aug 3, 2017
Priority date	Aug 3, 2016
Publication date	Jul 28, 2020
Grant date	Jul 28, 2020

Abstract

The disclosure provides compositions, kits and methods comprising an absorbent pad comprising a fibrous material and a liquid formulation comprising a luminescent substrate for identifying and quantifying biomolecule analytes.

First claim

What is claimed is:

1. A composition comprising:
a) an absorbent pad comprising (a) a fibrous material and (b) a liquid formulation which comprises a chemiluminescent substrate; wherein the pad: (i) has a thickness from 0.2 mm to 3 mm; (ii) has an extrinsic absorbency from 150 mL/m 2 to 500 mL/m 2 ; and (iii) is impregnated with the liquid formulation, wherein the fibrous material comprises a polyester/cellulose polymer blend, a hydroentangled polyester/cellulose blend, nylon, or a polyamide, and
b) a blot membrane in fluid contact with the absorbent pad, wherein the blot membrane comprises a biomolecule analyte bound to a detection agent, wherein the detection agent is reactive with the chemiluminescent substrate on the pad.

2. The composition of claim 1 , wherein the detection agent comprises an enzyme reactive with the chemiluminescent substrate.

3. The composition of claim 1 , wherein the absorbent pad has a basis weight from 25 g/m 2 to 100 g/m 2 .

4. The composition of claim 3 , wherein the absorbent pad has a basis weight from 50 g/m 2 to 70 g/m 2 .

5. The composition of claim 1 , wherein the absorbent pad has: (i) the thickness of the pad is from 0.2 mm to 1 mm; and (ii) the extrinsic absorbency of the pad is from 250 mL/m 2 to 400 mL/m 2 .

6. The composition of claim 5 , wherein the thickness of the pad is from 0.3 mm to 0.7 mm.

7. The composition of claim 1 , wherein the polyester/cellulose polymer blend comprises 40 wt % to 50 wt % polyester and 50 wt % to 60 wt % cellulose.

8. The composition of claim 1 , wherein the polyester/cellulose polymer blend comprises 45 wt % polyester and 55 wt % cellulose.

9. The composition of claim 1 , wherein the chemiluminescent substrate is a substrate for horseradish peroxidase, alkaline phosphatase, β-galactosidase, β-glucosidase, β-glucuronidase, arylesterase, sulfatase, or a combination of one or more thereof.

10. The composition of claim 1 , wherein the liquid formulation further comprises an oxidizing agent, a buffer, a stabilizing agent, an enhancer, or a combination of one or more thereof.

11. The composition of claim 10 , wherein the oxidizing agent is a peroxide compound, a compound that produces a peroxide in situ, hydrogen peroxide, urea hydrogen peroxide, sodium carbonate hydrogen peroxide, a perborate salt, or a combination of one or more thereof.

12. The composition of claim 10 , wherein the stabilizing agent is cyclodextrin, dextrin, a sulfate, a sugar, a nonionic surfactant, an anionic surfactant, an ethylene oxide/propylene oxide adduct, polyoxyethylenesorbitan monolaurate, polyoxyethylenesorbitan monopalmitate, polyethylene glycol sorbitan monostearate, polyethylene glycol sorbitan monooleate, polyoxyethylenesorbitan trioleate, t-octylphenoxypolyethoxyethanol, polyethylene glycol nonylphenyl ether, polyethylene glycol tert-octylphenyl ether, polyethylene glycol tert-octylphenyl ether, polyethylene glycol dodecyl ether, or a combination of one or more thereof.

13. The composition of claim 10 , wherein the buffer is ethylenediamine tetraacetic acid, succinate, citrate, aspartic acid, glutamic acid, maleate, cacodylate, 2-(N-morpholino)-ethanesulfonic acid, N-(2-acetamido)-2-aminoethanesulfonic acid, piperazine-N,N′-2-ethanesulfonic acid, 2-(N-morpholino)-2-hydroxy-propanesulfonic acid, N,N-bis-(hydroxyethyl)-2-aminoethanesulfonic acid, 3-(N-morpholino)-propanesulfonic acid, N-2-hydroxyethyl-piperazine-N-2-ethanesulfonic acid, 3-(N-tris-(hydroxymethyl)methylamino)-2-hydroxypropanesulfonic acid, 3-(N,N-bis[2-hydroxyethyl]amino)-2-hydroxypropanesulfonic acid, N-(2-hydroxyethyl)piperazine-N′-(2-hydroxypropanesulfonic acid), 4-(2-hydroxyethyl)-1-piperazine propanesulfonic acid, N-[tris(hydroxymethyl)-methyl]glycine, N,N-bis(2-hydroxyethyl)glycine, (2-hydroxy-1,1-bis(hydroxymethyl)ethyl)amino]-1-propanesulfonic acid, N-(1,1-dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid, tris(hydroxy methyl)amino-methane, and bis[2-hydroxyethyl]iminotris-[hydroxymethyl]methane, or a combination of one or more thereof.

14. The composition of claim 10 , wherein the enhancer is a halogenated phenol; an alkylated phenol; 4-benzylphenol; 4-(2′,4′-dinitrostyryl) phenol; 2,4-dichlorophenol; p-hydroxycinnamic acid; p-fluorocinnamic acid; p-nitroicinnamic acid; p-aminocinnamic acid; m-hydroxycinnamic acid; o-hydroxycinnamic acid; 4-phenoxyphenol; 4-(4-hydroxyphenoxy) phenol; p-phenylphenol; 2-chloro-4-phenylphenol; 4′-(4′-hydroxyphenyl) benzophenone; 4-(phenylazo) phenol; 4-(2′-carboxyphenylaza) phenol; 1,6-dibromonaphtho-2-ol; 1-bromonaphtho-2-ol; 2-naphthol; 6-bromonaphth-2-ol; 6-hydroxybenzothiazole; 2-amino-6-hydroxybenzothiazole; 2,6-dihydroxybenzothiazole; 2-cyano-6-hydroxybenzothiazole; dehydroluciferin; firefly luciferin; phenolindophenol; 2,6-dichlorophenolindophenol; 2,6-dichlorophenol-o-cresol; phenolindoaniline; a substituted or unsubstituted N-alkylphenoxazine; a substituted or unsubstituted N-alkylphenothiazine; a substituted or unsubstituted N-alkylpyrimidylphenoxazine; N-alkylpyridylphenoxazine; a substituted or unsubstituted 2-hydroxy-9-fluorenone; a substituted or unsubstituted 6-hydroxybenzoxazole, or a combination of one or more thereof.

15. The composition of claim 1 , wherein the chemiluminescent substrate is luminol, isoluminol, acridane, phenyl-10-methylacridane-9-carboxylate, 2,4,6-trichiorophenyl-10-methylacndane-9-carboxylate, pyrogallol, phloroglucinol, resorcinol, or a combination thereof.

16. A device system comprising: 1) a composition of claim 1 ; and 2) an imaging device.

17. A method for detecting a biomolecule analyte in a biological sample comprising:
(a) separating biomolecular components of a biological sample using gel electrophoresis thereby forming a separation gel comprising separated biomolecular components, wherein the separated biomolecular components comprise a biomolecule analyte;
(b) transferring the separated biomolecular components to a blot membrane;
(c) contacting the blot membrane with a detection agent and allowing the detection agent to bind to the biological analyte, wherein the detection agent is reactive with a luminescent substrate;
(d) contacting the blot membrane with an absorbent pad to provide the composition of claim 1 , wherein the luminescent substrate reacts with the detection agent thereby forming a luminescent signal; and
(e) detecting the luminescent signal thereby detecting the biomolecule analyte.

Assignees

• Pierce Biotechnology Inc

Inventors

• Desai Surbhi
• Severson Kari

Classifications

• C12Q1/28 Primary

  involving peroxidase · CPC title

• G01N33/54386

  Analytical elements · CPC title

• C09K11/06

  containing organic luminescent materials · CPC title

• F21K2/06

  using chemiluminescence · CPC title

• G01N21/76

  Chemiluminescence; Bioluminescence · CPC title