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KR-102952162-B1 - APPARATUS AND METHOD FOR SIMULTANEOUS SEPARATION OF CELLS AND MICROPARTICLES

KR102952162B1KR 102952162 B1KR102952162 B1KR 102952162B1KR-102952162-B1

Abstract

A microfluidic device is disclosed. The microfluidic device comprises: a sample chamber in which a biological sample is received and, by centrifugal force therein, the sample is separated into a first fluid layer containing a first target substance and a non-target substance and a second fluid layer containing a plurality of second target substances; a first separation module that receives a microparticle and receives the first fluid layer from the sample chamber to form a composite in which the microparticle and the non-target substance are combined, and separates the composite from the first target substance by a density difference; and a second separation module that receives the second fluid layer from the sample chamber and separates a plurality of second target substances by a density difference.

Inventors

  • 김민석
  • 이승준
  • 김종만

Assignees

  • 재단법인대구경북과학기술원
  • 주식회사 씨티셀즈

Dates

Publication Date
20260507
Application Date
20201201

Claims (10)

  1. In a microfluidic device mounted on a rotary drive unit that induces fluid flow by centrifugal force, A sample chamber that accommodates a biological sample, wherein the sample is separated by centrifugal force therein into a first fluid layer containing a first target substance and a non-target substance and a second fluid layer containing a plurality of second target substances; A first separation module that accommodates microparticles, receives the first fluid layer from the sample chamber to form a composite in which the microparticles and the non-target substance are combined, and separates the composite from the first target substance by a density difference; and A second separation module that receives the second fluid layer from the sample chamber and separates the plurality of second target substances by density difference; The above-mentioned first separation module is, A first reaction chamber connected to the sample chamber and in which the coupling is formed; A first separation chamber connected to the first reaction chamber and extending in a direction away from the center of rotation of the microfluidic device, wherein the aggregate is gathered at the lowest layer relative to the center of rotation of the microfluidic device due to a density difference; and It includes four recovery chambers connected to the first separation chamber and located at different distances from the rotation center of the microfluidic device, positioned staggered relative to the first separation chamber; The above second separation module is, A second reaction chamber connected to the sample chamber and formed by combining a first complex formed by combining with any one of the plurality of second target substances and a second complex formed by combining with another of the plurality of second target substances; and A second separation chamber connected to the second reaction chamber and extending in a direction away from the center of rotation of the microfluidic device, wherein the first coupling and the second coupling are separated by a density difference; It further includes a first recovery chamber and a second recovery chamber connected to the second separation chamber and arranged staggered relative to the second separation chamber, and A microfluidic device in which the first recovery chamber is located closer to the center of rotation than the second recovery chamber.
  2. In paragraph 1, The sample chamber and the first separation module are connected through a first channel having a first valve, and The sample chamber and the second separation module are connected through a seventh channel having a seventh valve, and Each of the first valve and the seventh valve melts when heated to open the first channel and the seventh channel, respectively. The above microfluidic device is, A microfluidic device configured to apply heat to each of the first valve and the second valve while mounted on the rotary drive unit and rotating, so that each of the first fluid layer and the second fluid layer can be moved to the first separation module and the second separation module while maintaining centrifugal force.
  3. In paragraph 1, The above sample is blood, and The first target substance is a circulating tumor cell (CTC), and The above-mentioned non-target substance is a white blood cell, and The above-mentioned microparticle is a microfluidic device that is a particle specifically binding to white blood cells.
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  5. In paragraph 3, A microfluidic device in which, among the four recovery chambers, the first recovery chamber closest to the center of rotation is for recovering a single CTC, the second recovery chamber located further from the center of rotation than the first recovery chamber is for recovering a platelet-CTC complex, the third recovery chamber located further from the center of rotation than the second recovery chamber is for recovering a leukocyte-CTC complex, and the fourth recovery chamber located between the third recovery chamber and the lowest layer is for recovering a CTC cluster.
  6. In paragraph 1, The above sample is blood, and The plurality of second target substances comprises at least a first exosome and a second exosome, and The second separation module accommodates a first microparticle that specifically binds to the first exosome to form a first complex and a second microparticle that specifically binds to the second exosome to form a second complex, wherein the first microparticle has a lower specific gravity than the second microparticle, a microfluidic device.
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  8. In paragraph 6, A microfluidic device in which the first recovery chamber is for recovering the first composite and the second recovery chamber is for recovering the second composite.
  9. In paragraph 1, The above sample is blood, and The plurality of second target substances comprises at least a first exosome and a second exosome, wherein the first exosome has a lower specific gravity than the second exosome, and The above second separation module is, Second separation chamber; A first recovery chamber connected to the second separation chamber and for recovering the first exosome; and A microfluidic device comprising: a second recovery chamber connected to the second separation chamber and for recovering the second exosomes.
  10. In Paragraph 9, The second separation module further includes a residue removal chamber connected to the second separation chamber, and A microfluidic device in which the second recovery chamber is located further from the center of rotation than the first recovery chamber, and the residue removal chamber is located closer to the center of rotation than the first recovery chamber.

Description

Apparatus and Method for Simultaneous Separation of Cells and Microparticles The present disclosure relates to a microfluidic device, and more specifically, to a microfluidic device for separating a target substance within a biological sample. Most deaths associated with malignant tumors are caused by metastasis to tissues and organs located away from the site where the tumor first originated. Therefore, early detection of metastasis is a critical determinant of survival for cancer patients, and early detection of tumors and monitoring of tumor growth are considered essential factors for the successful treatment of cancer patients. Circulating Tumor Cells (CTCs) refer to cancer cells that penetrate nearby blood vessels and circulate in the bloodstream to metastasize from the primary tumor, and are considered a key factor in understanding and diagnosing metastasis. Since CTCs exist in extremely small amounts at the 1 ppb level, it is very important to separate them with a high recovery rate and high purity. Previously, active research was conducted both domestically and internationally on various separation technologies, including antibody-based, filter, microfluidic, and density methods; however, due to the significant heterogeneity of CTCs, most studies were only able to isolate a limited number of CTCs. Furthermore, while fully automated technology is crucial for application in hospital settings, the development of high-quality separation equipment was lacking. FIG. 1 is a perspective view illustrating a microfluidic device according to an embodiment of the present disclosure, FIG. 2 is a configuration diagram of the interior of a microfluidic device according to an embodiment of the present disclosure, FIGS. 3 and 4 are drawings for explaining various embodiments of controlling a valve while maintaining centrifugal force without stopping the rotation of a microfluidic device, FIG. 5 is a schematic diagram illustrating an example of a recovery process of a first target substance (CTC) of a microfluidic device according to an embodiment of the present disclosure. FIG. 6 is a schematic diagram illustrating an example of a recovery process of a second target substance (exosomes) of a microfluidic device according to an embodiment of the present disclosure. FIG. 7 is a configuration diagram of the interior of a microfluidic device according to another embodiment of the present disclosure. Specific embodiments of the present disclosure will be described in detail below with reference to the drawings. However, the spirit of the present disclosure is not limited to the embodiments presented, and those skilled in the art who understand the spirit of the present disclosure may easily propose other embodiments that are inferior or included within the scope of the spirit of the present disclosure by adding, changing, or deleting other components within the same spirit, and such are also to be considered to be included within the spirit of the present disclosure. Terms used in this disclosure that are defined in general dictionaries may be interpreted as having the same or similar meaning as they have in the context of the relevant technology, and are not to be interpreted in an ideal or overly formal sense unless explicitly defined in this disclosure. In the absence of specific definitions, terms may be interpreted based on the overall content of this specification and the ordinary technical knowledge of the relevant field. Identical reference numbers or symbols in each attached drawing represent parts or components that perform substantially the same function. For convenience of explanation and understanding, the same reference numbers or symbols are used in different embodiments as well. That is, even if components having the same reference number are all depicted in multiple drawings, the multiple drawings do not represent a single embodiment. Additionally, in this disclosure, terms including ordinal numbers, such as "first," "second," etc., may be used to distinguish between components. These ordinal numbers are used to distinguish identical or similar components from one another, and the meaning of the terms should not be limited by the use of such ordinal numbers. For example, the order of use or arrangement of components combined with such ordinal numbers should not be restricted by the number. If necessary, each ordinal number may be used interchangeably. In this disclosure, singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, terms such as “comprising” or “consisting of” are intended to specify the presence of features, numbers, steps, actions, components, parts, or combinations thereof, and should be understood as not precluding the existence or addition of one or more other features, numbers, steps, actions, components, parts, or combinations thereof. The present disclosure relates to a microfluidic device, which is a device utilizing microfl