# US 10,768,182 B2 · Method for detecting nucleosomes containing histone modifications and variants

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- Number: US 10,768,182 B2
- Document ID: US10768182B2
- Status: Granted
- Kind: B2
- Application number: 15/768,981
- Filed: 2016-10-21
- Published: 2019-02-28
- Granted: 2020-09-08
- Earliest claimed: 2015-10-21
- Assignees: Belgian Volition SPRL
- Applicants: Belgian Volition Sprl
- Inventors: Mark Edward Eccleston; Jacob Vincent Micallef
- Web page: https://patentgrep.com/patent/US10768182B2

## CPC

- G01N 33/6875: {Nucleoproteins}
- G01N 33/57585: {involving compounds identifiable in body fluids}
- G01N 33/6893: {related to diseases not provided for elsewhere}
- 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 detecting and measuring the presence of cell free mono-nucleosomes and oligo-nucleosomes that contain histone H1 or a histone H1 modification, variant or isoform, and the use of such measurements for the detection and diagnosis of disease.

## Claims

### Claim 1

A method for detecting the presence of a cell free nucleosome containing histone H1 or a histone H1 modification, variant or isoform in a sample which comprises the steps of:
(i) contacting the sample with a first binding agent which binds to a non-histone H1 nucleosome epitope;
(ii) contacting the nucleosomes or sample with a second binding agent which binds to histone H1 or the histone H1 modification, variant or isoform;
(iii) detecting or quantifying the binding of said second binding agent to histone H1 or the histone H1 modification, variant or isoform in the sample; and
(iv) using the presence or degree of binding of said second binding agent as a measure of the presence of nucleosomes containing histone H1 or the histone H1 modification, variant or isoform in the sample.

### Claim 2

A method for detecting the presence of a cell free nucleosome containing histone H1 or a histone H1 modification, variant or isoform in a sample which comprises the steps of:
(i) contacting the sample with a first binding agent which binds to histone H1 or the histone H1 modification, variant or isoform;
(ii) contacting the nucleosomes or sample with a second binding agent which binds to a non-histone H1 nucleosome epitope;
(iii) detecting or quantifying the binding of said second binding agent to the non-histone H1 nucleosome epitope in the sample; and
(iv) using the presence or degree of binding of said second binding agent as a measure of the presence of nucleosomes containing histone H1 or the histone H1 modification, variant or isoform in the sample.

### Claim 3 (depends on 1)

A method as defined in claim 1, wherein the binding agent is an antibody.

### Claim 4 (depends on 1)

A method according to claim 1, wherein the sample is a body fluid.

### Claim 5 (depends on 1)

A method according to claim 1, wherein the sample is blood, serum or plasma.

### Claim 6 (depends on 2)

The method according to claim 2, wherein the binding agent is an antibody.

### Claim 7 (depends on 2)

The method according to claim 2, wherein the sample is a body fluid.

### Claim 8 (depends on 2)

The method according to claim 2, wherein the sample is blood, serum or plasma.