Process for preparing a biological sample

Patent metadata

Field	Value
Publication number	US-11905550-B2
Application number	US-201916585729-A
Country	US
Kind code	B2
Filing date	Sep 27, 2019
Priority date	Sep 28, 2018
Publication date	Feb 20, 2024
Grant date	Feb 20, 2024

Abstract

A process for preparing a biological sample including biological species, implemented in a preparation system, the preparation system including a device that includes: a housing, a first channel provided in the housing, a second channel provided in the housing, a chamber into which the first channel and the second channel open, a filter separating the chamber into two distinct spaces, the process including the following steps: injection of the biological sample in the form of a fluid via the first channel to concentrate biological species in the first space of the chamber of the device, injection of an immunological buffer fluid via the second channel of the device so as to at least partially elute the biological species with the immunological buffer fluid.

First claim

The invention claimed is:

1. A process for preparing a biological sample comprising biological species, implemented in a preparation system,
wherein the preparation system comprises a device comprising:
a housing comprising at least one opening,
a first channel provided in the housing,
a second channel provided in the housing,
a chamber into which the first channel and the second channel lead, wherein the chamber comprises a rough bearing surface arranged on the bottom of the chamber which extends over a majority of the bottom of the chamber,
a filter separating the chamber into two distinct spaces, wherein the first injection channel leads to a first space and the second channel leads to a second space, wherein the first space and the second space are partially superimposed such that the first space is a lower space in the chamber and is located under the filter and the second space is an upper space in the chamber and is located above the filter, such that the first space and the second space are in contact with each other in an superimposed area via the filter separating the first space and the and second space, wherein the filter has a porosity suitable for retaining the biological material to be analyzed,
the process comprising:
injecting the biological sample in the form of a fluid via the first channel to concentrate biological species in the first space of the chamber of the device,
conducting mechanical lysis of the biological species in the chamber by grinding the biological species by abrasion on the rough bearing surface, and separating cell membranes from an intracellular material,
eluting the intracellular material by injecting an amplification buffer through the first channel of the device, to elute the material via the second channel,
injecting an immunological buffer fluid via the second channel of the device and at least partially eluting the biological species with the immunological buffer fluid.

2. The process according to claim 1 , further comprising carrying out washing after concentration by injecting a wash buffer via the first channel of the device, thereby purifying the biological species.

3. A system for preparing a biological sample that comprises biological species, wherein the system comprises a device that comprises:
a housing comprising at least one opening,
a first channel provided in the housing,
a second channel provided in the housing,
a chamber into which the first channel and the second channel lead,
a filter separating the chamber into two distinct spaces, wherein the first injection channel leads to a first space and the second channel leads to a second space, wherein the first space and the second space are partially superimposed such that the first space is a lower space in the chamber and is located under the filter and the second space is an upper space in the chamber and is located above the filter, such that the first space and the second space are in contact with each other in an superimposed area via the filter separating the first space and the and second space, wherein the filter has a porosity suitable for retaining the biological material to be analyzed,
wherein the system comprises a fluid circuit connected to the device, wherein the fluid circuit comprises a plurality of controlled valves and a processing control unit configured to control the state of each valve to control the direction and flow of fluid within the system.

4. The system according to claim 3 , wherein the fluid circuit comprises a first valve arranged to connect a first end of the first channel of the device to an injection channel or to an immunological detection device.

5. The system according to claim 4 , wherein the fluid circuit comprises a second valve and three tanks and wherein the second valve is arranged to connect the injection channel to one of the three tanks.

6. The system according to claim 3 , wherein the fluid circuit comprises a third valve arranged to connect a first end of the second channel to an amplification detection device or to a third channel.

7. The system according to claim 6 , wherein the fluid circuit comprises a fourth valve arranged to connect the third channel to a tank to contain an immunoassay buffer or to a waste bin.

8. The system according to claim 3 , which is a system for the preparation of a biological sample for immunological detection.

9. The system according to claim 3 , which is a system for the preparation of a biological sample for detection by amplification.

10. The process according to claim 1 , wherein the grinding is carried out by a friction movement of the biological species against the rough bearing surface using a grinding member.

11. The process according to claim 10 , wherein the grinding member is a spatula or a rod, wherein the grinding member is applied from the exterior of the chamber and its end is applied against an external surface of a membrane to stretch the membrane and the filter towards the bottom of the chamber and to rub the biological species present in a sample against the rough bearing surface, and
wherein the membrane is located towards the top of the chamber above the second space, and wherein an upper wall of the housing has an opening sealed by the membrane such that the membrane is anchored in the housing.

12. The process according to claim 1 , wherein the housing comprises a membrane located towards the top of the chamber and above the second space, wherein an upper wall of the housing has an opening sealed by the membrane such that the membrane is anchored in the housing.

13. The process according to claim 1 , wherein the rough bearing surface arranged on the bottom of the chamber has an average surface roughness parameter of from 0.1 μm to 10 μm.

14. The process according to claim 13 , wherein the average surface roughness parameter of the rough bearing surface is from 0.2 μm and 3 μm.

Assignees

• Commissariat Energie Atomique
• Service Dept Metropolitain Dincendie Et De Secours Sdmis

Inventors

• Bourdat Anne-Gaelle
• Wenisch Gregory

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• B01L3/502746

  characterised by the means for controlling flow resistance, e.g. flow controllers, baffles or throttle valves · CPC title

• B01L3/502753

  characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation · CPC title

• C12N15/1017

  by filtration, e.g. using filters, frits, membranes · 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