# US 2019/0170741 A1 · Method for detecting and/or characterising tumour cells and associated apparatus

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- Number: US 2019/0170741 A1
- Document ID: US20190170741A1
- Status: Published application
- Kind: A1
- Application number: 16/093,934
- Filed: 2017-04-18
- Published: 2019-06-06
- Earliest claimed: 2016-04-15
- Assignees: Chu Montpellier; Ecole Superieure De Physique Et De Chimie Industrielles De La Ville De Paris; Centre National De La Recherche Scientifique; Universite De Montpellier; Axlr, Satt du Languedoc Roussillon
- Applicants: Chu Montpellier; Ecole Superieure De Physique Et De Chimie Industrielles De La Ville De Paris; Centre national de la recherche scientifique; Université de Montpellier; AxLR, SATT du Languedoc Roussillon
- Inventors: Catherine Michèle Marilou ALIX - PANABIERES; Andrew David GRIFFITHS; Raphaël Clément Li-Ming DOINEAU; Clément NIZAK; Philippe Chi-Thanh NGHE; Jean-Marie Pierre BAUDRY; Elodie Michèle Christine BRIENT-LITZLER; Klaus EYER; Jérôme BIBETTE
- Web page: https://patentgrep.com/patent/US20190170741A1

## CPC

- G01N 33/54333: {Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction}
- B01L 3/502784: {specially adapted for droplet or plug flow, e.g. digital microfluidics}
- G01N 33/532: Production of labelled immunochemicals
- G01N 33/5436: {with ligand physically entrapped within the solid phase (liposomes G01N33/5432; immunological test elements G01N33/54386)}
- G01N 33/57595: {involving intracellular compounds}
- B01L 2200/0673: Handling of plugs of fluid surrounded by immiscible fluid
- 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

The present invention relates to the field of biological diagnosis in oncology. It relates to a method and apparatus for detecting and/or characterizing tumor cells by detecting one or more elements of the tumor cell secretome, in particular one or more peptides or proteins, and, in particular, one or more tumor markers. The invention also relates to detecting and/or characterizing droplets of tumor cells and their method of preparation.

## Claims

### Claim 1

Method for detecting and/or characterizing tumor cells (90), comprising:
the supply of a plurality of droplets (6) contained in a carrier fluid (8), at least one of the droplets (6) comprising at least one aggregate (10) of particles (12) defining an elongated object along a main axis (X), at least some droplets (6) containing a cell (90) capable of producing at least one target element of the secretome of a tumor cell capable to be fixed on the aggregate;
the measurement of at least one physical parameter characteristic of the fixing of a target element (37) of the secretome of a tumor cell on the aggregate (10); and
the detection and/or the characterization of tumor cells (90) from the measurement of the at least one physical parameter.

### Claim 2 (depends on 1)

Method according to claim 1, wherein the method is a method for detecting and/or characterizing single tumor cells comprising:
the supply of a plurality of droplets (6) contained in a carrier fluid (8), at least one of the droplets (6) comprising at least one aggregate (10) of particles (12) defining an elongated object along a main axis (X), at least some droplets (6) containing a single cell (90);
the incubation of the single-cell-containing (90) droplets (6) under conditions and for a time sufficient for the single cell (90) being capable to produce at least one target element (37) of the secretome of a tumor cell that is capable to be fixed to the aggregate (10);
the measurement of at least one physical parameter, each physical parameter being characteristic of the fixing of a distinct target element (37) from the secretome of a tumor cell on the aggregate (10); and
the detection and/or characterization of single tumor cells (90) from the measurement of the at least one physical parameter.

### Claim 3 (depends on 1)

Method according to claim 1, wherein the cell (90) is derived from a biological fluid and is selected from the group consisting of a circulating tumor cell (CTC) and a disseminated tumor cell (DTC).

### Claim 4 (depends on 1)

Method according to claim 1, wherein the measuring step comprises the measurement of the at least one physical parameter locally at a plurality of points in the droplet (6), wherein the measurement step preferably comprises the determination of the integral of the measured values within the droplet (6).

### Claim 5 (depends on 1)

Method according to claim 1, wherein the measurement step is performed in a microfluidic chamber without circulation of the droplets.

### Claim 6 (depends on 1)

Method according to claim 1 comprising:
providing a device comprising a set for putting in circulation (22) the droplet and a plurality of classification zones (94, 96), and a means (98) for selectively directing the droplet or a portion of the droplet towards a classification zone (94, 96),
the decision of classifying the droplet (6) or a portion of the droplet, the decision consisting in selectively choosing one of the plurality of classification zones (94, 96),
the transport of the droplet (6), respectively of a portion of the droplet, to the classification zone of the droplet (6) chosen in the decision step.

### Claim 7 (depends on 1)

Method according to claim 1, wherein at least one droplet comprises at least two distinct signaling entities (34), wherein each of the two signaling entities (34) is able to form a complex with a distinct target element (37) on the aggregate (10), wherein the method comprises measuring a signal indicating the concentration of each of the relocated signaling entities (34).

### Claim 8 (depends on 7)

Method according to claim 7, wherein at least some droplets (6) comprise a tumor cell capable of secreting, cleaving or releasing one or more elements of the tumor cell secretome, wherein each element of the tumor cell secretome is a distinct target element (37), wherein the measurement of the signal indicates the concentration of each of the relocated signaling entities (34) thus allowing quantification of the element(s) of the tumor cell secretome.

### Claim 9

Apparatus for detecting and/or characterizing tumor cells (90) comprising:
a set for supplying a plurality of droplets (6) contained in a carrier fluid (8), wherein at least one of the droplets (6) comprises at least one aggregate (10) of particles (12) defining an object elongated according to a main axis (X), wherein at least some droplets (6) contain a tumor cell (90) capable of producing a target element (37) capable to be fixed on the aggregate (10),
characterized in that the apparatus comprises a set for measurement of a physical parameter characteristic of the fixing of a target element (37) on the aggregate (10), wherein the apparatus optionally comprises, in addition:
a set for putting in circulation the droplet (22),
a set for classification decision of the droplet,
a set of sorting the droplet according to the classification decision.

### Claim 10

Droplet for detecting and/or characterizing tumor cells (90), comprising a plurality of particles capable of forming an aggregate (10) of particles (12) defining an elongated object along a main axis (X), and a tumor cell (90), and optionally at least one target element (37) of the secretome of the tumor cell (90) intended to be fixed on the aggregate (10).

### Claim 11

Method for preparing droplets (6) for detecting and/or characterizing tumor cells, comprising:
the dispersion, in a mass of fluid (86) intended to form droplets (6), of particles (12) capable of forming an aggregate (10) defining an object elongated along a main axis (X), and of a plurality of cells (90), wherein at least some cells (90) are tumor cells capable of producing a target element (37) of the secretome of a tumor cell, then
the dispersion of the mass of fluid (86) in the form of droplets (32), so that each droplet comprises a plurality of particles (12), while at least some of the droplets (6) comprise, in addition, a cell (90), and
optionally forming in each droplet (6) at least one aggregate (10) of particles (12) defining an object elongated along a main axis (X), wherein the aggregate (10) of particles (12) is formed in each droplet (6) after the dispersion.