# US 8,846,328 B2 · Method for detecting the presence of a protease enzyme in a sample

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- Number: US 8,846,328 B2
- Document ID: US8846328B2
- Status: Granted
- Kind: B2
- Application number: 13/733,438
- Filed: 2013-01-03
- Published: 2013-07-25
- Granted: 2014-09-30
- Earliest claimed: 2006-02-23
- Assignees: Mologic Limited
- Applicants: Mologic Limited
- Inventors: Paul J. Davis; Mark J. Davis; Mark Burnapp; Sandra Hemmington
- Web page: https://patentgrep.com/patent/US8846328B2

## CPC

- G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)}
- B01L 3/5055: {Hinged, e.g. opposable surfaces}
- B01L 2300/069: Absorbents; Gels to retain a fluid
- B01L 2300/0809: rectangular shaped
- B01L 2300/0887: Laminated structure
- 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)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

An enzyme detection product for detecting the presence of an enzyme in a sample. The product includes: a reaction zone for receiving the sample; a visualization zone for presenting a signal in response to the detection of the activity of the enzyme; and a membrane. The membrane is interposable between the reaction zone and the visualization zone and prevents passage from the reaction zone to the visualization zone the components having a size greater than a threshold size. The reaction zone includes a reactant capable of reacting with the enzyme in order to generate a reaction product having a size less than a threshold size.

## Claims

### Claim 1

A method of detecting the presence of a protease in a sample comprising the steps of:
i) contacting the sample with a first reactant, the reactant being a peptide capable of being cleaved by the protease and producing a reaction product in the presence of the protease, wherein said reaction product is detectable as a consequence of cleavage by the protease;
ii) separating at least some of the components in the sample which are smaller than a threshold size into a visualisation zone, the reaction product being smaller than the threshold size; wherein said separating comprises using a selective membrane interposed between the reaction zone and visualization zone to separate components in the sample, the membrane preventing passage from the reaction zone to the visualization zone of components having a greater size than the threshold size; and
iii) detecting the presence of the reaction product in the visualisation zone in order to indicate the presence of the protease in the sample.

### Claim 2 (depends on 1)

A method according to claim 1 wherein step ii) further comprises observing the reaction product or a signal released by the reaction product.

### Claim 3 (depends on 1)

A method according to claim 1 wherein step i) is carried out on a reaction substrate and step iii) is carried out on a visualisation substrate.

### Claim 4 (depends on 3)

A method according to claim 3 and further comprising the step of, after step i), contacting the reaction substrate with the visualisation substrate.

### Claim 5 (depends on 1)

A method according to claim 1 wherein step iii) further comprises providing a second reactant, which reacts with the reaction product to generate a signal in order to indicate the presence of the protease in the sample.