# US 9,435,815 B2 · Method for rapid detection of hepatocyte growth factor in biological fluids

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- Number: US 9,435,815 B2
- Document ID: US9435815B2
- Status: Granted
- Kind: B2
- Application number: 14/380,153
- Filed: 2013-02-22
- Published: 2015-02-05
- Granted: 2016-09-06
- Earliest claimed: 2012-02-22
- Assignees: Peas Institut AB
- Applicants: PEAS Institut AB
- Inventors: Fariba Nayeri; Kamal Bahar
- Web page: https://patentgrep.com/patent/US9435815B2

## CPC

- G01N 33/74: involving hormones {or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors}
- G01N 33/52: Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)}
- G01N 2333/4753: Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present invention relates to a method for determining the presence, absence or amount of biologically active HGF in a sample, comprising the steps (i) bringing the sample in contact with a first porous solid phase comprising a HGF binding component of the extracellular matrix or cell membrane, and an indicator composition comprising bromothymol blue and a quaternary ammonium compound; and (ii) correlating the color of the porous solid phase with the presence, absence or amount of biologically active HGF in the sample. It further relates to a device comprising a first porous solid phase comprising at least one HGF-binding component of the extracellular matrix or cell membrane, and an indicator composition comprising bromothymol blue and a quaternary ammonium compound.

## Claims

### Claim 1

A method for determining the presence, absence or amount of biologically active Hepatocyte Growth Factor (HGF) in a fecal sample of a subject and correlating said presence, absence or amount of HGF with an acute inflammation of the bowel in the subject, said method comprising the steps of:
bringing the fecal sample in contact with a first and a second porous solid phase of a test strip, said first and second porous phases being fixed to a solid support, said first porous phase comprising at least one HGF binding component of the extracellular matrix or cell membrane, and an HGF indicator composition in contact with the HGF binding component, said HGF indicator composition comprising bromothymol blue and a quaternary ammonium compound, said second porous phase comprising a pH indicator composition fixed to said second porous phase, said pH indicator composition comprising bromothymol blue, a quaternary ammonium compound and methyl red; and
correlating the colour of the first porous solid phase with the color of a reference solution containing a known quantity of biologically active HGF and thereby determining presence, absence or amount of biologically active HGF in the fecal sample and determining the pH of the fecal sample from said second solid phase, wherein the presence of an amount or biologically active HGF in said fecal sample is indicative of an acute inflammation of the bowel in the subject when the pH of said fecal sample is >8 and wherein said HGF binding component is heparin sulphate proteoglycan or dextran sulphate, wherein the fecal sample is a fluid.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the correlation of the colour of the porous solid phase with the presence, absence or amount of biologically active HGF in the sample includes comparing the colour of the porous solid phase with a porous solid phase that has been brought into contact with at least one reference solution of known HGF content.

### Claim 3

A test strip comprising a solid support, a first porous solid phase fixed on solid support and a second porous solid phase fixed on said solid support, said first porous solid phase comprising at least one HGF-binding component of the extracellular matrix or cell membrane, wherein said component is heparan sulphate proteoglycan or dextran sulphate, and an HGF indicator composition in contact with said component, said HGF indicator composition comprising bromothymol blue and a quaternary ammonium compound, said second porous solid phase comprising a pH indicator composition, said pH indicator composition fixed to said second porous solid phase, said pH indicator composition comprising bromothymol blue, a quaternary ammonium compound and methyl red.

### Claim 4 (depends on 3)

The test strip according to claim 3, wherein at least one of said first and second porous solid phases comprises cellulose.