WO2024090849A1 - Cartouche pour dispositif de séparation centrifuge et dispositif de séparation centrifuge la comprenant - Google Patents

Cartouche pour dispositif de séparation centrifuge et dispositif de séparation centrifuge la comprenant Download PDF

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Publication number
WO2024090849A1
WO2024090849A1 PCT/KR2023/015601 KR2023015601W WO2024090849A1 WO 2024090849 A1 WO2024090849 A1 WO 2024090849A1 KR 2023015601 W KR2023015601 W KR 2023015601W WO 2024090849 A1 WO2024090849 A1 WO 2024090849A1
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WO
WIPO (PCT)
Prior art keywords
chamber
cartridge
target
cartridges
cartridge body
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Application number
PCT/KR2023/015601
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English (en)
Korean (ko)
Inventor
이성훈
길민석
배진한
Original Assignee
주식회사 클리노믹스
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Application filed by 주식회사 클리노믹스 filed Critical 주식회사 클리노믹스
Publication of WO2024090849A1 publication Critical patent/WO2024090849A1/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
    • B04B—CENTRIFUGES
    • B04B7/00—Elements of centrifuges
    • B04B7/08—Rotary bowls
    • B04B7/12—Inserts, e.g. armouring plates
    • B04B7/16—Sieves or filters
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
    • B04B—CENTRIFUGES
    • B04B15/00—Other accessories for centrifuges
    • B04B15/06—Other accessories for centrifuges for cleaning bowls, filters, sieves, inserts, or the like

Definitions

  • the present invention relates to a cartridge structure that can perform centrifugal separation of specific components from a large volume of body fluid, such as blood, and extract and separate target substances within a single cartridge.
  • This application is the result of the market-friendly global competitiveness product development project [Project identification number: 1711174434 (Project number RS-2020-KD000019)], a project supported by the Pan-Ministry Life Cycle Medical Device Research and Development Project, and is a hospital information system (HIS) )
  • the application was filed by claiming priority for Korean Patent Application No. 10-2022-0139833, which was filed as a result of the development of a real-time liquid biopsy molecular diagnosis system for diagnosing linked lung cancer.
  • the rotary circular or polygonal Lab-on-a-Disk technology involves putting whole blood into a single tube container to separate it into each component (plasma, buffy coat, white blood cells, and red blood cells). This is a method that allows efficient separation without cross-contamination of samples compared to separation methods based on specific gravity by inserting them into a centrifugal separation device.
  • cell-derived nucleic acid which has recently been widely used in cancer diagnosis, exists in the plasma layer, and in order to separate it, it must be separated manually using a filter or magnetic beads.
  • the present invention was devised to solve the above-mentioned problem, and the purpose of the present invention is to extract a chamber for cell lysis and a target component within a cartridge for a centrifugal separator capable of separating and analyzing body fluid (e.g., whole blood) components.
  • a cartridge structure that provides a magnetic bead reaction chamber for the reaction and at the same time places a target material recovery chamber within the cartridge, enabling sample separation and efficient extraction of target components in a single centrifugal separation cartridge without separate manual work. is to provide.
  • a disk-shaped cartridge body 100 having a constant thickness that can be rotated by receiving power from a drive unit;
  • Separated component storage module (A) including a plurality of chambers for storing separated components centrifuged by receiving body fluid through the inlet of the cartridge body (A): disposed inside the cartridge body (A) and storing the separated components
  • An extraction module (B) that lyses the components introduced from the module (A) and then extracts the target components through a magnetic bead reaction; a cleaning fluid module (C) that communicates with a part of the chamber of the extraction module (B) and has at least one cleaning fluid supply chamber for supplying a cleaning fluid;
  • a chamber for cell lysis and a magnetic bead reaction chamber for extracting target components are provided in a cartridge for a centrifugal separator capable of separating components of body fluid (e.g., whole blood), and at the same time, target substances.
  • body fluid e.g., whole blood
  • the cartridge of the present invention with this structure, even a skilled technician can accurately and efficiently separate target components (e.g., cell-derived nucleic acid, cfDNA) from a sample (e.g., whole blood) in one device without cross-contamination.
  • target components e.g., cell-derived nucleic acid, cfDNA
  • a sample e.g., whole blood
  • cell-derived nucleic acid can be extracted quickly without cross-contamination, and such cell-derived nucleic acid contains various genetic information, especially cell-derived nucleic acid derived from cancer cells.
  • cfDNA contains genetic information about cancer cells and can be useful in cancer diagnosis.
  • FIG. 1 and 2 show a plan cross-sectional conceptual diagram of a cartridge for a centrifugal separator according to an embodiment of the present invention.
  • Figure 3 shows an image of a triple disk type used in a centrifugal separation device using three of the above-described cartridges of the present invention.
  • Figures 4 to 7 show the results of testing the application performance of a cartridge for a centrifugal separator according to an embodiment of the present invention.
  • Cartridge body A Separate component storage module
  • Impurity storage module 110 First chamber part
  • Figures 1 and 2 show a plan cross-sectional conceptual diagram of a cartridge for a centrifugal separator (hereinafter referred to as 'the present invention') according to an embodiment of the present invention.
  • the present invention includes a disk-shaped cartridge body 100 having a constant thickness (t) that can be rotated by receiving power from a driving unit, and receiving body fluid through the injection port of the cartridge body (A).
  • a separation component storage module (A) including a plurality of chambers for storing centrifuged components, disposed inside the cartridge body (A), and magnetically lysing the components flowing from the separation component storage module (A).
  • An extraction module (B) that extracts the target component through a bead reaction
  • a cleaning fluid module (C) having at least one cleaning fluid supply chamber that communicates with a part of the chamber of the extraction module (B) and supplies a cleaning fluid
  • the extraction module It selectively communicates with (B) through centrifugal force and a valve, and may be configured to include an impurity storage module (D) that stores cleaning liquid and impurities other than the target ingredients.
  • the cartridge body 100 is implemented as a three-dimensional structure with a plurality of chambers inside and passages between each chamber, as shown in Figure 1.
  • the cartridge body 100 A flat cover structure is provided at the top and bottom of the top and bottom to seal the three-dimensional structure from top and bottom.
  • the cartridge body 100 injects body fluid, which is an object requiring separation and analysis, into the interior, and implements chamber structures such as a chamber for cell lysis, a reaction chamber for separation, and a cleaning fluid supply chamber embedded within the cartridge body.
  • Each chamber structure is implemented as a closed structure, and material movement between them is realized by centrifugal force and the opening and closing operation of the valve.
  • the cartridge structure of the present invention is applied to a device that performs centrifugation by injecting body fluid, and the various body fluids to be analyzed in the present invention may be whole human blood, sweat, or other samples, but the present invention
  • human whole blood is used as the target body fluid
  • the first separated material is plasma
  • the target component is cell free DNA (cfDNA).
  • the cartridge body 100 may be implemented as an external shape having a fan-shaped flat cross-section with a constant thickness, as shown in FIGS. 1 and 2, and in the present invention, the center (P1) of this external shape is separated from the left and right sides.
  • a structure defined as the intersection of line segments extending the outer border of and the angle a formed by the center P1 is 120° will be described as a preferred embodiment.
  • the injection port 1 is formed closest to the center P1 of the present invention, and the chamber parts that perform the separation function for body fluids can be sequentially arranged around the injection port outward.
  • the injection port 1 is a place where body fluid (whole blood) is injected, and is the part where the body fluid separated by the present invention by performing a centrifugation operation is first injected.
  • the separated component storage module (A) includes a plurality of chambers that receive body fluid through an injection port and store the separated components centrifuged, and the separated material is stored.
  • the separate component storage module (A) includes a first chamber portion 110, an adjacent chamber 115, an outer chamber 117, and a micro-channel 11 that communicates them.
  • the extraction module (B) performs the function of lysing the components introduced from the separated component storage module (A) and then extracting the target component through a magnetic bead reaction.
  • the extraction module (B) receives the first isolate by centrifugal force from the first chamber unit 110, which separates and stores the first isolate containing the target ingredient, and collects the first isolate (plasma).
  • a second chamber unit 120 that dissolves the inner cells and separates them into a target component and a second separation material. In the second chamber unit, the dissolved target component is introduced, and the magnetic beads and the target component react to form the target component. It may be configured to include a third chamber part 130 that separates and a fourth chamber part 140 that stores the target material flowing from the third chamber part.
  • the extraction module (B) extends from the center (P1) of the cartridge body 100 to the outside (P2), and includes the first chamber part 110, the second chamber part 120, and the third chamber part 130. ) and the fourth chamber portion 140 are arranged sequentially. That is, in FIG. 2, considering an arbitrary straight line P2 extending from the center P1 to the outside, each chamber portion includes a first chamber portion 110, a second chamber portion 120, and a third chamber portion 130. ) and the fourth chamber part 140 are arranged in that order, and this arrangement is to ensure smooth transfer of the separated material according to centrifugal force.
  • the movement of materials between the first chamber part 110, the second chamber part 120, the third chamber part 130, and the fourth chamber part 140 in communication with each other occurs in the flow path according to the opening and closing of the valve. It opens and moves selectively, and is implemented by centrifugal force resulting from the rotation of the cartridge body (A).
  • the second chamber 120 disposed outside the first chamber 110 is selectively communicated with the first chamber 110 by the valve 14, During the centrifugation process, the plasma can be moved to the second chamber 120 through the fine connection passage 121 by controlling the opening and closing of the valve 14 and centrifugal force.
  • the second chamber 120 performs the function of lysing cells in plasma by receiving lysing substances necessary for lysing the injected cells.
  • the dissolved material injected into the second chamber 120 may be, for example, a Protease K solution.
  • the lysed material and the plasma reflect each other to perform cell lysis. For example, to completely achieve cell lysis, centrifugation at 180 degrees can be performed in a centrifugal separator.
  • the third chamber 130 is disposed adjacent to the outside of the second chamber 120.
  • the magnetic bead provided in the third chamber 130 Cell-derived nucleic acid (cf DNA) reacts through a reaction with beads. That is, in the third chamber, magnetic beads and cell-derived nucleic acid (cfDNA) in dissolved plasma are combined.
  • the present invention allows a washing process to be performed to remove impurities present in the plasma present in the third chamber.
  • the cleaning fluid module (C) for performing this cleaning process includes at least one cleaning fluid chamber disposed on one side of the extraction module (B), and the cleaning fluid chamber 160 includes the third chamber portion 130 and The cleaning solution is supplied through communication to remove impurities in the first separating component.
  • the cleaning fluid module (C) may be configured to further include at least one cleaning fluid supply chamber 160 that communicates with the third chamber portion 130 and supplies cleaning fluid.
  • the cleaning liquid supply chamber 160 of the present invention is used to remove impurities present in plasma before and after the reaction of plasma flowing into the magnetic beads performed in the third chamber 130.
  • the cleaning fluid supply chamber 160 is provided with a total of four cleaning fluid chambers (161, 162, 163, 164), and can supply a total of four cleaning fluids to perform four cleaning processes. Let it happen.
  • the necessary cleaning solution is injected and filled in advance before centrifugation, and when centrifugation begins, the cleaning solution is sequentially supplied to the third chamber 130 by centrifugal force, the opening and closing operation of the valves 16, 17, 18, and 19, and centrifugal force. This movement removes impurities.
  • the removed impurities are moved to the impurity storage chamber (D) by the opening and closing operation of the valve 23 and centrifugal force.
  • the cell-derived nucleic acid (cfDNA) bound to the magnetic beads in the third chamber 130 is combined with the extraction buffer solution and dissolved.
  • This extraction buffer solution is injected through the extraction buffer solution inlet 12, and is injected into the third chamber 130 by the opening and closing operation of the valve 21 and centrifugal force, and the cells combined with the magnetic beads are injected.
  • the derived nucleic acid is dissolved.
  • Cell-derived nucleic acid (cfDNA) dissolved in the elution buffer flows into the fourth chamber 140 disposed outside the third chamber 130 by centrifugal force and the opening and closing operation of the valve 29.
  • the bottom surface of the third chamber 130 of the present invention is disposed adjacent to the fourth chamber portion. It is made to have a first inclination ( ⁇ 1) inclined toward the outside where (140) is located.
  • the first inclination ⁇ 3 is preferably in the range of 3° to 5° based on the horizontal plane of the floor. If the angle exceeds the above-mentioned range, the force with which the target material flows along the slope increases due to centrifugal force, making it difficult to achieve stability, and if the slope is less than 3°, residual target ingredients remain, making it difficult to achieve the desired effect. do.
  • Cell-derived nucleic acid (cfDNA) collected in the fourth chamber 140 can be recovered using pellets.
  • Figure 3 shows an actual use image of a triple disk type in which three of the above-described cartridges of the present invention are applied to a centrifugal separation device.
  • target components e.g., cell-derived nucleic acid, cfDNA
  • a sample e.g., whole blood
  • work efficiency can be improved by more than 3 times.
  • Figures 4 to 7 show the extraction concentration of intracellular nucleic acid (cfDNA) from 120 lung cancer patients at Asan Medical Center in Seoul, where clinical blood was collected from the patients and whole blood samples from randomly selected patients were collected. This is compared to the one released using the existing manual method.
  • cfDNA intracellular nucleic acid
  • concentrations of intracellular nucleic acid (cfDNA) separated using the cartridge according to the embodiment of the present invention were measured on average at 596 pg/ul and 691 pg/ul, respectively, and the concentration of intracellular nucleic acid (cfDNA) using the existing manual method was It was measured at 439pg/ul.
  • cell-derived nucleic acid can be extracted quickly without cross-contamination, and such cell-derived nucleic acid contains various genetic information, especially cell-derived nucleic acid derived from cancer cells.
  • cfDNA contains genetic information about cancer cells and can be useful in cancer diagnosis.

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  • Centrifugal Separators (AREA)

Abstract

La présente invention concerne une structure de cartouche qui peut centrifuger des composants spécifiques à partir de fluides corporels tels qu'un grand volume de sang, et peut extraire et séparer une substance cible à l'intérieur d'une seule cartouche. La structure de cartouche comprend une chambre pour la lyse cellulaire et une chambre de réaction de bille magnétique pour extraire des composants cibles, dans la cartouche, et en même temps, une chambre de récupération de substance cible est placée à l'intérieur de la cartouche pour mettre en œuvre une séparation d'échantillon et une extraction efficace d'un composant cible dans une seule cartouche de séparation centrifuge même sans travail manuel supplémentaire.
PCT/KR2023/015601 2022-10-27 2023-10-11 Cartouche pour dispositif de séparation centrifuge et dispositif de séparation centrifuge la comprenant WO2024090849A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020220139833A KR20240059058A (ko) 2022-10-27 2022-10-27 원심분리장치용 카트리지 및 이를 포함하는 원심분리 장치
KR10-2022-0139833 2022-10-27

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WO2024090849A1 true WO2024090849A1 (fr) 2024-05-02

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101228308B1 (ko) * 2007-05-23 2013-01-31 삼성전자주식회사 미세유동 칩을 이용한 디스크형 미세유동장치 및 생체물질마이크로어레이 칩을 이용한 디스크형 미세유동장치
KR101973604B1 (ko) * 2017-07-21 2019-04-29 서강대학교산학협력단 무세포 유전물질(cell free tumor DNA)을 검출하기 위한 미세유체장치 및 검출방법
KR20210106837A (ko) * 2020-02-21 2021-08-31 주식회사 클리노믹스 원심 분리 장치용 챔버 및 이를 포함하는 원심 분리 장치
KR20210138640A (ko) * 2019-03-12 2021-11-19 노비럭스 엘엘씨 현장 진단 농도 분석기
KR102404003B1 (ko) * 2020-06-30 2022-05-31 주식회사 클리노믹스 미세 유동 장치

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102063865B1 (ko) 2018-06-20 2020-01-08 울산과학기술원 원심력 기반 혈소판 분리 및 검진 장치

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101228308B1 (ko) * 2007-05-23 2013-01-31 삼성전자주식회사 미세유동 칩을 이용한 디스크형 미세유동장치 및 생체물질마이크로어레이 칩을 이용한 디스크형 미세유동장치
KR101973604B1 (ko) * 2017-07-21 2019-04-29 서강대학교산학협력단 무세포 유전물질(cell free tumor DNA)을 검출하기 위한 미세유체장치 및 검출방법
KR20210138640A (ko) * 2019-03-12 2021-11-19 노비럭스 엘엘씨 현장 진단 농도 분석기
KR20210106837A (ko) * 2020-02-21 2021-08-31 주식회사 클리노믹스 원심 분리 장치용 챔버 및 이를 포함하는 원심 분리 장치
KR102404003B1 (ko) * 2020-06-30 2022-05-31 주식회사 클리노믹스 미세 유동 장치

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