# US 10,481,166 B2 · Microparticle fractionation

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- Number: US 10,481,166 B2
- Document ID: US10481166B2
- Status: Granted
- Kind: B2
- Application number: 15/970,952
- Filed: 2018-05-04
- Published: 2018-09-06
- Granted: 2019-11-19
- Earliest claimed: 2011-11-30
- Assignees: Singapore Health Services Pte Ltd; Agency for Science, Technology and Research (A*Star)
- Applicants: Agency for Science, Technology and Research; Singapore Health Services Pte Ltd.
- Inventors: Sai Kiang Lim; Kok Hian Tan
- Web page: https://patentgrep.com/patent/US10481166B2

## CPC

- G01N 33/689: {related to pregnancy or the gonads}
- G01N 33/5091: {for testing the pathological state of an organism}
- G01N 33/6872: {Intracellular protein regulatory factors and their receptors, e.g. including ion channels}
- G01N 33/6893: {related to diseases not provided for elsewhere}
- G01N 2333/47: Assays involving proteins of known structure or function as defined in the subgroups
- G01N 2405/10: Glycosphingolipids, e.g. cerebrosides, gangliosides
- G01N 2800/32: Cardiovascular disorders
- G01N 2800/368: Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour
- G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
- G01N 2800/7028: Cancer
- 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

We describe a method of monitoring the state of a cell, tissue, organ or organism. The method comprises establishing, for a sample of micro-particles from the cell, tissue, organ or organism, a ratio. The ratio is of a selected polypeptide in microparticles which comprise GM1 gangliosides, preferably which bind to Cholera Toxin B (CTB) (“GM1 ganglioside microparticle polypeptide”) to the selected polypeptide in microparticles which comprise exposed phos—photidylserine, preferably which bind to Annexin V (“Annexin V microparticle polypeptide”). The GM1 ganglioside microparticle polypeptide to Annexin V microparticle polypeptide ratio so established may be indicative of the state of the cell, tissue, organ or organism.

## Claims

### Claim 1

A method for selecting microparticles in a sample, the method comprising selecting microparticles by size; and one or both of:
(a) selecting microparticles in the sample which comprise GM1 gangliosides; and
(b) selecting microparticles in the sample which comprise exposed phosphotidylserine.

### Claim 2 (depends on 1)

The method of claim 1, in which step (a) comprises selecting microparticles in the sample which bind to Cholera Toxin subunit B (CTB); or in which step (b) comprises selecting microparticles in the sample which bind to Annexin V, or both.

### Claim 3

A method for selecting microparticles in a sample, the method comprising:
(a) selecting microparticles in the sample which comprise GM1 gangliosides; and/or
(b) selecting microparticles in the sample which comprise exposed phosphotidylserine;
wherein the microparticles comprise CD9+ microparticles.

### Claim 4 (depends on 3)

The method of claim 3, in which step (a) comprises selecting microparticles in the sample which bind to Cholera Toxin subunit B (CTB); or in which step (b) comprises selecting microparticles in the sample which bind to Annexin V, or both.

### Claim 5 (depends on 3)

The method of claim 3, wherein the sample is selected from the group consisting of urine, blood, tears, saliva, bronchoaveolar fluid, tumoral effusions, epididymal fluid, amniotic fluid and milk.

### Claim 6 (depends on 3)

The method of claim 3, wherein the microparticles comprise microvesicles, exosomes, ectosomes or apoptotic bodies.

### Claim 7 (depends on 1)

The method of claim 1, which comprises both selecting steps (a) and (b).

### Claim 8

A method for selecting microparticles in a sample, the method comprising:
(a) selecting microparticles in the sample which comprise GM1 gangliosides; and
(b) selecting microparticles in the sample which comprise exposed phosphotidylserine.