WO2012057495A2

WO2012057495A2 - Metal screen filter

Info

Publication number: WO2012057495A2
Application number: PCT/KR2011/007974
Authority: WO
Inventors: 전병희; 이종길
Original assignee: 주식회사 싸이토젠
Priority date: 2010-10-25
Filing date: 2011-10-25
Publication date: 2012-05-03
Also published as: WO2012057495A3

Abstract

The present invention relates to a metal screen filter for separating cancer cells from blood. The metal screen filter of the present invention is formed with a plurality of filtration holes for filtering cancer cells, such that the gap between two neighboring filtration holes of the plurality of filtration holes is formed to be 45 - 65 % the diameter of a cancer cell, and is installed into a passage having blood flow containing both cancer cells and non-target cells. The diameter of a filtration hole for filtering a cancer cell is 5 - 25 ㎛. According to the present invention, cancer cells can be efficiently and easily separated and collected from large amounts of blood. Additionally, the collection rate of live cancer cells is improved by smooth emission of cancer cells out of a catheter by flow of solution.

Description

Metal screen filter

The present invention relates to a metal screen filter, and more particularly to a metal screen filter for separating cancer cells from the blood (Blood).

Recently, regulations on animal tests and clinical trials for treating human diseases have been tightened. In order to replace these animal and clinical trials, research and technology are being actively developed to collect live cells from human blood. Cell collection is performed by various cell collection devices such as microfluidic devices, CTC chips, circulating tumor cells chips, and filters.

As an example of a cell collection device, US Patent Application Publication No. 2007 / 0025883A1 discloses a Parylene membrane filter for filtering cells from a fluid. The membrane filter is mounted in a chamber and has a plurality of pores that are formed to prevent passage of cells, for example, cancer cells. However, in order to collect and collect the cancer cells filtered in the membrane filter from the chamber, a solution such as water must be supplied in the chamber in a reverse direction to force the cancer cells out of the chamber. It is very difficult to collect and collect, and cancer cells are easily damaged.

As another example of a cell collection device, US Patent Application Publication No. 2009 / 0188864A1 discloses a method and apparatus for microfiltration to perform cell separation. A plurality of filter patches are installed in the central square hole of the microfiltration device. Filter patches consist of a membrane having a plurality of pores for filtration of cells. However, there is a problem that it is difficult to collect and collect the cells filtered in the filter patch of the microfiltration device from the central square hole, and the cells are likely to be damaged.

The present invention is to solve various problems of the conventional cell collection device as described above. An object of the present invention is to provide a metal screen filter capable of efficiently separating cancer cells from blood.

Another object of the present invention is to provide a metal screen filter which can improve the collection rate by smoothly discharging cancer cells out of the conduit by the flow of the solution.

Still another object of the present invention is to provide a metal screen filter capable of preventing clogging of the filtration hole by non-target cells in blood.

According to one aspect of the invention, a metal screen filter is provided. The metal screen filter according to the present invention is provided in a passage through which blood containing cancer cells and non-target cells flows, and a plurality of filtration holes are formed for filtration of cancer cells, and two adjacent ones of the plurality of filtration holes are formed. The gap between the filtration holes is formed to 45-65% of the diameter of the cancer cells. In addition, the interval is formed in 4 ~ 21㎛, the antibody surface layer is formed on the inner surface of the plurality of filter holes for capturing cancer cells.

Metal screen filter according to another aspect of the present invention, is installed in the blood flow path containing cancer cells and non-target cells, a plurality of filtration holes having a diameter of 5 ~ 25㎛ for filtration of cancer cells are formed have. In addition, the diameter of the plurality of filtration holes is formed in 7 ~ 21㎛, the inner surface of the plurality of filtration holes are coated with an antibody surface layer for capturing cancer cells.

The metal screen filter according to the present invention can easily and efficiently separate and collect cancer cells from a large amount of blood. In addition, the flow of the solution can smoothly discharge the cancer cells out of the conduit to improve the collection rate of live cancer cells. In addition, since clogging of the filtration holes by non-target cells is prevented, it can be usefully employed for analysis, testing, drug testing, clinical trials, and the like of cancer cells.

1 is a cross-sectional view showing a configuration in which a metal screen filter according to the present invention is installed in a conduit.

Figure 2 is a perspective view showing the configuration of a metal screen filter according to the present invention.

Figure 3 is a perspective view showing the configuration of a metal screen filter according to the present invention.

4 is a perspective view partially showing the configuration of the metal screen filter according to the present invention.

5 is a partial cross-sectional view for explaining the filtering process of cancer cells by a metal screen filter according to the present invention.

Figure 6 is a partial cross-sectional view for explaining the process of collecting cancer cells from the metal screen filter according to the present invention.

7 is a perspective view showing the configuration of another embodiment of a metal screen filter according to the present invention.

8 is a perspective view partially cut out the configuration of a metal screen filter of another embodiment according to the present invention;

9 is a cross-sectional view showing the configuration of a metal screen filter of another embodiment according to the present invention.

Other objects, specific advantages and novel features of the present invention will become more apparent from the following detailed description and preferred embodiments associated with the accompanying drawings.

Hereinafter, preferred embodiments of the metal screen filter according to the present invention will be described in detail with reference to the accompanying drawings.

First, referring to FIG. 1, a metal screen filter 10 according to the present invention is installed in a passage 22 of a conduit 20 for transporting blood 2. The blood 2 is a target cell for filtering by the metal screen filter 10 and includes a plurality of cancer cells 4 and a plurality of non-target cells 6. Non-target cells 6 are composed of red blood cells (Red blood cells) and white blood cells (White blood cells).

1 to 5, the metal screen filter 10 according to the present invention is installed in the passage 22 of the conduit 20 for transporting the blood 2 to filter the cancer cells 4. The metal screen filter 10 has a surface 12, a back surface 14 and a plurality of filtration holes 16 formed for filtration of the cancer cells 4. 2 and 3, the metal screen filter 10 is shown to be formed in a circular shape, which is an example, the shape of the metal screen filter 10 is a square, oval, rectangular, etc. according to the shape of the passage (22) Can be changed as appropriate.

The filtration holes 16 are formed to have a size that the cancer cells 4 cannot pass through for filtration of the cancer cells 4. That is, the size of the filtration holes 16 is formed smaller than the diameter of the cancer cells (4). 2 to 4 show that the filtration holes 16 are formed in a quadrangular shape, but the shape of the filtration holes 16 may be appropriately changed into a circle, an ellipse, a rectangle, and the like as necessary. When the filtration holes 16 are formed in a circular shape, the diameters of the filtration holes 16 are formed to be 5 ~ 25㎛.

The gap G between two adjacent filtration holes 16 among the filtration holes 16 is formed to be narrower than the diameter of the cancer cells 4. The gap G between the two filtration holes 16 is formed at 45 to 65% of the diameter of the cancer cells 4. The metal screen filter 10 may be made of, for example, stainless steel, nickel, aluminum, or copper. The filtration holes 16 having a micrometer size for filtration of the cancer cells 4 are formed by etching or electroforming using MEMS (Micro-electro mechanical systems). Can be formed. This metal screen filter 10 is not deformed by the pressure of the blood 2 or the solution flowing through the passage 22.

Referring to FIG. 1, blood 2 is supplied upstream of conduit 20 by blood supply means. The blood supply means may be, for example, in various forms such as a syringe (Syringe), a blood collection tube, a bag, a pack that can store a quantity of blood and supply it upstream of the conduit 20. Can be configured. In addition, the blood supply means may be composed of a syringe pump (Syringe pump), a plunger pump (Plunger pump) and the like.

1 and 5, the blood 2 flows along the passage 22 of the conduit 20 and passes through the filtration holes 16 and then is discharged out of the conduit 20. Cancer cells 4 in the blood 2 do not pass through the filtration holes 16 and remain on the surface 12. Non-target cells 6, ie red blood cells and white blood cells, which have a higher strain than cancer cells 4, pass easily through the filtration holes 16. Leukocytes vary in shape and size, depending on the shape and number of nuclei, and the presence or absence of granules. White blood cells easily pass through the filtration holes 16 of a size smaller than the diameter by the deformation of the cytoplasm.

Referring to FIG. 6, after filtration of the cancer cells 4, the cancer cells 4 may be discharged out of the passage 22 by supplying a solution 8 in the reverse direction or the forward direction to the passage 22. The solution 8 is supplied in the reverse direction to minimize the damage of the cancer cells 4. The solution 8 may be supplied by syringe, syringe pump, plunger pump or the like. The solution may consist of a diluent that dilutes the blood (2), water, a diluent acid such as Triton X-100 or nitric acid. The cancer cells 4 discharged out of the passage 22 by the supply of the solution 8 can be collected by simply collecting them in a container, for example, a test tube, a culture dish, or the like.

On the other hand, cancer cells having a diameter of 7.5-15 μm pass through a tube having a diameter of 8 μm when a pressure of about 100 mmHg is applied. Cancer cells with a diameter of 15 μm passing through a 8 μm diameter tube have a strain of about 53%. Back washing, i.e., when the solution flows in the direction opposite to the flow of the blood 2, and discharges the cancer cells 7.5 µm in diameter out of the passage 22, considering the strain of the cancer cells, the two filtration holes 16 The interval G between them is preferably about 4 μm or less.

Non-small cell lung cancer and breast cancer are known to have a diameter of about 40㎛ the cancer cells. In consideration of the strain of the cancer cells having a diameter of 40 μm during back washing, the distance G between the two filtration holes 16 is preferably set to about 21 μm or less. When cancer cells having a diameter of 7.5 μm are present between two filtration holes 16 larger than the distance G of 4 μm, the cancer cells may not be dropped from the surface 12 as they are deformed by the flow of the solution. In addition, cancer cells having a diameter of 40 μm may not fall off the surface 12 between the two filtration holes 16 larger than the gap G 21 μm.

In the metal screen filter 10 according to the present invention, the gap G between the two filtration holes 16 is 45-65% of the diameter of the cancer cells in consideration of the pressure of the solution. Cancer cells having a diameter of 7.5 μm may not fall off the surface 12 between the two filtration holes 16 by backwashing when the gap G exceeds 65%, ie, greater than about 4.9 μm. Cancer cells with a diameter of 40 μm may not fall off the surface 12 between the two filtration holes 16 by backwashing when the gap G exceeds 65%, ie, more than 26 μm. Increasing the pressure of the solution to forcibly drop the cancer cells stuck to the surface 12 between the two filtration holes 16 causes damage to the cancer cells, thereby lowering the collection rate of living cancer cells. If the gap G for cancer cells having a diameter of 7.5 μm is less than 45%, that is, less than about 3.375, the metal screen filter 10 may be damaged by the flow of the blood 2 and the solution.

As clearly shown in FIGS. 4 and 5, an anti-body surface layer 30 is formed on the inner surface of the filtration holes 16 for the capture of the cancer cells 4. The antibody of the antibody surface layer 30 may be, for example, an anti-Epithelial Cell Adhesion Molecule antibody (Anti-EpCAM antibody), an anti-Cytokeratin antibody, an Anti-CK antibody, or the like. Can be configured. As shown in FIG. 5, when the smaller cancer cells 4a of the cancer cells 4 pass through the filter holes 16, the cancer cells 4a are made of either an Anti-EpCAM antibody or an Anti-CK antibody. It is bound to the antibody surface layer 30 which consists of, and is captured. Therefore, the metal screen filter 10 according to the present invention can efficiently collect and collect cancer cells 4 contained in a large amount of blood.

7 to 9 show another embodiment of a metal screen filter according to the present invention. 7 to 9, the metal screen filter 110 of another embodiment is installed in the passage of the conduit like the metal screen filter 10 described above. The metal screen filter 110 of another embodiment has a surface 112, a back side 114 and a plurality of filtration holes 116 formed for filtration of the cancer cells 4.

The filtration holes 116 are arranged in a regular pattern. The diameter of the filtration holes 116 is formed smaller than the diameter of the cancer cells (4). The diameter of the filtration holes 116 is formed of 5 ~ 25㎛. Preferably, the diameter of the filtration holes 116 may be formed to 7 ~ 25㎛. The antibody surface layer 130 is coated on the inner surface of the filtration holes 116 to capture the cancer cells 4. Although the filter holes 116 are illustrated in FIG. 8 in a circular shape, the shape of the filter holes 116 may be appropriately changed into a rectangle, an ellipse, a rectangle, and the like as necessary. When the filter holes 116 are formed in a rectangular or rectangular shape, the spacing between two adjacent filter holes 116 is formed at 45 to 65% of the diameter of the cancer cells 4.

As shown in FIG. 9, blood 2 flows along the passage of the conduit, passes through the filtration holes 116, and then exits the conduit. Cancer cells 4 in the blood 2 do not pass through the filtration holes 116 and remain on the surface 112. Cancer cells 4 have a lower strain than erythrocytes and leukocytes, and thus cannot pass through filtration holes 116 of diameter smaller than that.

Non-target cells 6 pass through filtration holes 116. As an example of the non-target cells 6, the red blood cells have a diameter of about 7.2 to 8.4 µm. Human red blood cells have a concave disc shape in the center and have no nucleus. Red blood cells have a high strain rate and easily pass through the filtration holes 116 having a diameter of 5 μm. When the diameters of the filtration holes 116 are formed to be 7 탆 in order to filter the cancer cells 4 having a diameter of 7 탆 or more, the red blood cells smoothly pass through the filtration holes 116. Therefore, clogging of the filtration holes 116 due to the red blood cells, which exist about 5 million blood per liter, is prevented, and the flow of the blood 2 is maintained smoothly.

As another example of the non-target cells (6), white blood cells vary in shape and size, depending on the shape and number of nuclei, and the presence or absence of granules, and are about 10 to 21 μm in diameter. White blood cells easily pass through the filtration holes 116 of a diameter smaller than that by the deformation of the cytoplasm. The largest macrophage of leukocytes (Macrophage) is 21 μm in diameter. When the diameter of the filter holes 116 is formed to 20㎛, most of the white blood cells present in the 7,000 ~ 8,000 per 1 ㎣ of blood passes through the filter holes (116). When the small cancer cells 4a of the cancer cells 4 pass through the filter holes 116, the cancer cells 4a are bound to the antibody surface layer 130 made of either an Anti-EpCAM antibody or an Anti-CK antibody. Is captured.

The embodiments described above are merely illustrative of the preferred embodiments of the present invention, the scope of the present invention is not limited to the described embodiments, those skilled in the art within the spirit and claims of the present invention It will be understood that various changes, modifications, or substitutions may be made thereto, and such embodiments are to be understood as being within the scope of the present invention.

Claims

It is installed in the blood flow passage containing cancer cells and non-target cells, and a plurality of filtration holes are formed for filtration of the cancer cells, and the distance between two adjacent filtration holes of the plurality of filtration holes Metal screen filter is formed in 45 ~ 65% of the diameter of the cancer cells.
The method of claim 1,

The gap is formed of 4 ~ 21㎛ metal screen filter is formed narrower than the diameter of the non-target cells.
The method of claim 1,

The diameter of the plurality of filter holes is a metal screen filter is formed in 5 ~ 25㎛.
The method according to any one of claims 1 to 3,

Metal surface filter is formed on the inner surface of the plurality of filter holes for the antibody surface layer to capture the cancer cells.
A metal screen filter is provided in the passage through which blood containing cancer cells and non-target cells, and a plurality of filtration holes having a diameter of 5 ~ 25㎛ for filtration of the cancer cells.
The method of claim 5,

And a diameter of the plurality of filtration holes is 7 ~ 21㎛.
The method of claim 5,

And a gap between two adjacent filtration holes of the plurality of filtration holes is 45 to 65% of the diameter of the cancer cell.
The method according to any one of claims 5 to 7,

Metal screen filter is coated on the inner surface of the plurality of filtration holes for the antibody surface layer to capture the cancer cells.