Device for lysing biological species and method implemented by said device

Patent metadata

Field	Value
Publication number	US-10160947-B2
Application number	US-201515313740-A
Country	US
Kind code	B2
Filing date	May 27, 2015
Priority date	May 28, 2014
Publication date	Dec 25, 2018
Grant date	Dec 25, 2018

Abstract

A device and method for lysing biological species present in a fluid includes implementing a rough surface against which the objects to be lysed are crushed by a shearing motion. A device for mechanically lysing biological species has a first and second wall mounted movable relative to each other, between an initial position in which the walls are separated from each other, and a lysing position in which the first wall presses against the second wall. The first and second walls also are mounted shearingly movable relative to each other in the lysing position. At least one of the first or second walls has a rough bearing surface against the other wall and has a mean surface roughness parameter Ra of between 0.2 μm and 10 μm, and preferably between 0.2 μm and 3 μm.

First claim

The invention claimed is:

1. A device for the mechanical lysis of a biological species contained in a fluid, comprising:
a first wall and a second wall mounted movable relative to one another between an initial position in which the first and second walls are drawn apart and a lysis position in which the first wall presses against the second wall, the first and second walls also being mounted movable under shear relative to one another in the lysis position, wherein at least one of the first and second walls has a rough bearing surface against the other wall and has an average surface roughness parameter Ra of between 0.2 μm and 10 μm, or between 0.2 μm and 3 μm.

2. The mechanical lysis device according to claim 1 , wherein one of the first and second walls is a microporous wall porous to liquids and non-porous to the biological species to be lysed.

3. The mechanical lysis device according to claim 2 , wherein the microporous wall has a mean pore diameter of less than 1 μm, or less than 0.2 μm.

4. The mechanical lysis device according to claim 2 , wherein the microporous wall is rough.

5. The mechanical lysis device according to claim 4 , wherein the rough microporous wall is formed of a layer of microporous material having a planar surface on which a rough layer is positioned intended to press against the other wall in the lysis position.

6. The mechanical lysis device according to claim 4 , wherein the rough microporous wall is formed of a layer of microporous material having a rough surface.

7. The mechanical lysis device according to claim 4 , wherein the rough microporous wall is formed of a layer of macroporous material having a rough surface on which a macroporous layer is positioned intended to press against the other wall in the lysis position, the microporous layer being sufficiently thin and flexible to follow the contour of the surface roughness of the layer in the macroporous material in the lysis position.

8. The mechanical lysis device according to claim 2 , wherein the microporous wall is not rough and has an average surface roughness parameter Ra of less than 0.1 μm, whereas the other and non-microporous wall is rough and has an average surface roughness parameter Ra of between 0.2 μm and 10 μm, preferably between 0.2 μm and 3 μm.

9. The mechanical lysis device according to claim 4 , wherein the first wall and the second wall are rough.

10. The mechanical lysis device according to claim 1 , further comprising a chamber and a piston mounted mobile in the chamber between the initial position and the lysis position, the first wall being mounted on the mobile piston and the second wall being arranged facing the first wall.

11. The mechanical lysis device according to claim 10 , wherein the second wall is microporous porous to liquids and non-porous to the biological species to be lysed, and wherein the chamber also comprises a fluid outlet arranged opposite the first wall relative to the second microporous wall.

12. The mechanical lysis device according to claim 11 , wherein the chamber also comprises a fluid inlet and another fluid outlet.

13. The mechanical lysis device according to claim 12 , wherein the chamber also comprises an impervious membrane arranged between the first wall, the fluid inlet and fluid outlet, the membrane being sufficiently thin and flexible to follow the contour of the roughness of at least one among the first and second walls in the lysis position.

14. A method for the mechanical lysis of a biological species comprising the following steps:
A) Providing a device for the mechanical lysis of a biological species contained in a fluid, comprising a first wall and a second wall mounted movable relative to one another between an initial position in which the first and second walls are drawn apart and a lysis position in which the first wall presses against the second wall, the first and second walls also being mounted movable under shear relative to one another in the lysis position, wherein at least one of the first and second walls has a rough bearing surface against the other wall and has an average surface roughness parameter Ra of between 0.2 μm and 10 μm, or between 0.2 μm and 3 μm so that the first and second walls lie in the initial position;
B) Placing the biological species to be lysed between the first and second walls;
C) Bringing the first and second walls to the lysis position;
D) Generating shear between the first and second walls in the lysis position to cause mechanical lysis of the biological species and to obtain a lysate;
E) Drawing the first and second walls apart until they reach the initial position; and
F) Recovering the lysate of the biological species.

15. The method for mechanical lysis of a biological species according to claim 14 , wherein:
at step A) the device further comprises a chamber and a piston mounted mobile in the chamber between the initial position and the lysis position, the first wall being mounted on the mobile piston and the second wall being arranged facing the first wall;
at step C) the piston is caused to slide inside the chamber until the first and second walls are in the lysis position; and
at step D) the piston is rotated so that the first and second walls remain in contact with each other.

16. The method for mechanical lysis of a biological species according to claim 15 , wherein:
at step A) the second wall is microporous porous to liquids and non-porous to the biological species to be lysed, and the chamber also comprises a fluid outlet arranged opposite the first wall relative to the second microporous wall;
at step B) a liquid containing the biological species to be lysed is placed between the first and second walls;
at step C) the piston is caused to slide inside the chamber until the liquid has been filtered through the microporous wall, solely the biological species to be lysed are left remaining on the microporous wall and the first and second walls are in the lysis position; and
at step D) the piston is rotated so that the first and second walls remain in contact with each other.

17. The method for mechanical lysis of a biological species according to claim 16 , wherein:
at step A) the chamber also comprises a fluid inlet and another fluid outlet;
at step B) the liquid containing the biological species to be lysed is caused to enter through the fluid inlet between the first and second walls;
at step C) the piston is caused to slide inside the chamber until the first and second walls are in the lysis position;
at step D) the piston is rotated so that the first and second walls remain in contact with each other; and
at step F) an eluting liquid is circulated between the fluid inlet and the fluid outlet to recover the lysate through the fluid outlet.

18. The method for mechanical lysis of a biological species according to the preceding claim 17 , wherein:
at step A) the chamber also comprises an impervious membrane arranged between the first wall, the fluid inlet and fluid outlet, the membrane being sufficiently thin and flexible to follow the contour of the roughness of at least one among the first and second walls in the lysis position;
at step B) a liquid containing the biological species to be lysed is caused to enter through the fluid inlet between the impervious membrane and the second wall;
at step C) the piston is caused to slide inside the chamber until the first and second walls are in the lysis position via the impervious membrane;
at step D) the piston is rotated so that the first and second walls remain in contact with each other via

Assignees

• Commissariat Energie Atomique

Inventors

• Bourdat Anne-Gaelle
• Viana Antonio
• Den Dulk Remco

Classifications

• C12M39/00

  Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms (apparatus for cleaning laboratory receptacles or instruments B01L13/02; cleaning in general B08B) · CPC title

• C12M47/06

  Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material (lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003) · CPC title

• C12N1/066 Primary

  by physical processes · CPC title

• C12M33/00

  Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus (chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00) · CPC title

• G01N1/286

  involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F) · CPC title