KR102073249B1 - Membrane-based Devices for Separating Plasma from Blood - Google Patents

Abstract

Embodiments of the present disclosure provide a membrane based device capable of selectively separating only plasma from a blood (specifically whole blood) sample using a membrane filter.

Description

Membrane-based Devices for Separating Plasma from Blood

The present disclosure relates to a device for separating plasma from blood. More specifically, the present disclosure relates to a chip type device capable of selectively separating only plasma from blood (specifically whole blood) samples using membrane filters.

Blood circulating in the blood vessels of the human body or various animals is a liquid flowing in the blood vessels and contains various components such as red blood cells, white blood cells, platelets, and plasma. The blood carries oxygen received from the lungs to tissue cells, releases carbon dioxide from the tissues in vitro, and transports nutrients absorbed from the digestive tract to organs and tissue cells, as well as discharges various unnecessary components out of the body. In addition, the hormone secreted by the endocrine gland is carried to the target organs and tissues, the body temperature is kept constant, and in addition, it has various functions such as killing various bacteria, viruses, etc. that have invaded the living body.

On the other hand, what should be preceded in the prevention and treatment of the disease is the diagnosis of the disease, various tests or diagnostic methods are applied according to the characteristics of the individual disease. Representative disease screening methods include methods such as computed tomography and magnetic resonance images to visually check for health problems, and obtain biological tissue suspected of having a problem. This method is used to directly check whether the virus is infected or deformation of the biological tissue, or indirectly confirms the infection by taking blood or tissue fluid and analyzing its components.

Among them, the identification of diseases through analysis of blood has the advantage of being the cheapest, simplest, and shorter time, and is widely used as a test method that is primarily performed before other methods. Blood consists of red blood cells, white blood cells and platelets, of which about 42-47% of whole blood is solid, while the remaining 53-58% consists of plasma, which is a liquid component. Each blood cell has about 5 million red blood cells, 8000 white blood cells, and about 300,000 platelets in 1 μl of blood. Plasma is a viscous liquid with about 90% water content, the remainder consisting of proteins, carbohydrates, fats, etc., containing trace vitamins, enzymes, hormones, antibodies, electrolytes, and the like.

When blood cells are used during the examination of the disease through whole blood, the number, form, size, and amount of hemoglobin are checked in a manner that contrasts with normal cases, and the amount and ratio of various components even when using plasma The disease is diagnosed in a way that contrasts with normal.

On the other hand, plasma contains a variety of nutrients, enzymes, hormones, antibodies and the like, there is a wide range of diseases that can be diagnosed through this. For example, diseases that can be diagnosed with plasma tests include diabetes, hypothyroidism, liver disease, pregnancy, atherosclerosis, myocardial infarction, acute hepatitis, anemia, muscle trauma, mucus edema, viruses, drug addiction, typhoid fever, measles, rubella, etc. As extensive. As such, the types of diseases that can be diagnosed through plasma are more diverse than when using blood cells.

However, red blood cells in whole blood are easily destroyed in the analysis process, and exposure of components in blood cells serves as a barrier to ensuring the accuracy of diagnosis. Therefore, in order to increase the reliability of the diagnosis, it may be desirable to use only plasma from which blood cells and the like have been removed from the whole blood. In particular, a protein chip is a kind of biochip used to detect the presence or absence of a specific protein contained in a blood sample or to detect a specific protein amount and to diagnose a disease related to a protein. Since the protein to be detected is mainly present in plasma, the protein chip is a protein chip. In order to obtain a highly sensitive and quantitative result, it is necessary to separate only plasma components from blood.

As such, studies to separate plasma from whole blood are continuously conducted. Thus, conventional methods used to separate plasma from whole blood may be classified into two types depending on whether energy sources are used.

As an active particle separation method, the centrifugal separation method can be illustrated typically. Centrifugal separation is a method of separating the components constituting the mixture according to the difference in density by centrifugal force generated by rotating the vessel containing the mixture at high speed.

As a passive separation method, a filter, a capillary tube, etc. are typically used. A filter is a method in which separation is performed by arranging a structure in a path through which particles for separation move, and allowing particles larger than the space of the structure to be captured by the structure. Through this process, the mixed solvent and particles may be separated or particles may be separated according to their size (for example, Korean Patent No. 1773509, WO2004 / 084974, etc.). The capillary method uses the principle of separating the non-particle solution from the mixed solution. Unlike the above-described method, the solution and the particles are separated from the mixed solution by controlling the behavior of the non-particle solution (eg, domestic Patent No. 0889727, etc.).

In addition, various methods have been proposed to separate plasma from small amounts of whole blood, for example, by placing microstructures smaller than the blood cells in the flow path and pumping blood from the outside, the blood cells are filtered by the microstructures to extract only the plasma. By placing a low-level diaphragm that prevents blood cells from passing through and leaving only plasma components through the diaphragm; placing paper, glass fibers, porous media, or membranes on the side or front of the blood flow Separation methods, a method of layering blood cells and plasma by layering using the sedimentation effect of blood cells by gravity, and extracting only plasma, and deflecting the flow of blood cells by applying an electrical signal are also known.

The filter method, which is most widely used among the above-described passive plasma separation methods, has a disadvantage in that the space for plasma separation is large and space utilization is low, and the separation efficiency is low because the flow of blood and plasma is very slow in the plasma separation process. In contrast, the present inventors have developed a device that improves plasma separation efficiency by promoting the flow of separated blood (Domestic Patent No. 1602949). However, the above-described technique is easy to manufacture the plasma separation device is not only composed of a three-layer structure of the upper substrate, the intermediate substrate and the lower substrate, but also the overall structure is complicated, such as a structure of various patterns are required for the individual substrates Rather, it is further constrained to combine or automate in a modular fashion with other devices.

Therefore, while overcoming the limitations of the prior art and suppressing the outflow of blood cells, hemolysis and the like, plasma can be separated and recovered from whole blood with high efficiency, while having a simple structure for easy production. The demand is increasing.

In one embodiment of the present disclosure has a simple structure compared to the prior art to facilitate the manufacture, and to provide a chip-type device capable of quickly separating plasma components even if a small amount of whole blood is added.

In addition, one embodiment of the present disclosure seeks to provide a chip-type device for separating plasma from whole blood having a structure suitable for a modular and / or automated manner.

According to one aspect of the present disclosure,

An upper structure including at least one first opening formed to inject a blood sample, and a second opening spaced apart from the first opening and connected to the negative pressure applying means at one end side region; And

(i) a membrane filter seat configured to seat a membrane filter for separating plasma from a blood sample injected through the first opening, (ii) a plasma chamber for collecting plasma separated from and in communication with the second opening, and (iii) an undercarriage comprising a first plasma transfer passage defined by the bottom surface, the plasma separated by the membrane filter extending to move in the direction of the second opening;

Including,

Here, a blood chamber is formed to receive the blood sample injected between the upper structure and the lower structure by assembling the upper structure and the lower structure,

The membrane filter seating portion defines a width of a first plasma movement passage in a central region of a bottom surface formed by a pair of surfaces inclined symmetrically or asymmetrically downward from both sides of the substructure,

The first plasma migration passage is in communication with the plasma chamber by connecting means, and

On the pair of symmetrically or asymmetrically inclined surfaces of the membrane filter seating portion, the first guide protrusion pattern and the second guide protrusion protrude so that the plasma separated through the membrane filter moves while converging to the first plasma movement path by capillary action. A device for separating plasma from blood is provided, each of which is provided with a pattern.

According to one embodiment of the present disclosure,

An upper structure including a first opening formed in one end side region for injecting a blood sample, and a second opening positioned in an end side region in a direction opposite to the first opening and connected to the sound pressure applying means; And

(i) a membrane filter seat configured to seat a membrane filter for separating plasma from a blood sample injected through the first opening, (ii) a plasma chamber for collecting plasma separated from and in communication with the second opening, and (iii) a lower structure comprising a first plasma migration passage inclined downwardly and bounded by a bottom surface such that plasma separated by the membrane filter moves in the direction of the second opening;

Including,

Here, the blood chamber is formed to accommodate the blood sample injected between the upper structure and the lower structure by the assembly of the upper structure and the lower structure,

The membrane filter seating portion defines a width of a first plasma movement passage in a central region of a bottom surface formed by a pair of surfaces inclined symmetrically or asymmetrically downward from both sides of the substructure,

The first plasma migration passage is in communication with the plasma chamber by connecting means, and

On the pair of symmetrically or asymmetrically inclined surfaces of the membrane filter seating portion, the first guide protrusion pattern and the second guide protrusion protrude so that the plasma separated through the membrane filter moves while converging to the first plasma movement path by capillary action. A device for separating plasma from blood is provided, each of which is provided with a pattern.

According to an exemplary embodiment, the upper structure may further include an air outlet.

According to an exemplary embodiment, the first opening may be a circular cavity, and the cavity may be formed in a fused shape of the cavity and the air outlet.

According to an exemplary embodiment, the first opening may be a circular cavity, and the cavity and the air outlet may be formed separately.

According to an exemplary embodiment, the first opening may have a shape in which air outlets are fused in a wing shape on both sides of the circular cavity.

According to another embodiment of the present disclosure,

A pair of first openings having a shape extending in the longitudinal direction to inject a blood sample, and a second opening formed in an end side region in a direction opposite to the first openings and connected to the sound pressure applying means; Superstructure; And

(i) a membrane filter seat configured to seat a membrane filter for separating plasma from a blood sample injected at least once through the first opening; and (ii) a plasma collecting plasma separated from and communicated with the second opening. A lower structure including a chamber and (iii) a first plasma migration passage extending to horizontally move plasma separated by the membrane filter in the direction of the second opening and bounded by a bottom surface;

Including,

Here, a blood space is formed to accommodate the blood sample injected between the upper structure and the lower structure by the assembly of the upper structure and the lower structure,

The membrane filter seating portion defines a width of a first plasma movement passage in a central region of a bottom surface formed by a pair of surfaces inclined symmetrically or asymmetrically downward from both sides of the substructure,

The first plasma migration passage is in communication with the plasma chamber by connecting means, and

On the pair of symmetrically or asymmetrically inclined surfaces of the membrane filter seating portion, the first guide protrusion pattern and the second guide protrusion protrude so that the plasma separated through the membrane filter moves while converging to the first plasma movement path by capillary action. A device for separating plasma from blood is provided, each of which is provided with a pattern.

According to an exemplary embodiment, the upper structure is provided with a support for fixing the membrane filter protruding while being spaced apart at a predetermined interval on the back, so that the membrane filter is in close contact with the membrane filter seat when assembled with the lower structure. And to enhance the capillary phenomenon in the second guide protrusion pattern.

According to an exemplary embodiment, the upper structure and the lower structure may each be a polymer material.

According to another aspect of the present disclosure,

a) mounting a membrane filter on the aforementioned device;

b) taking a blood sample and injecting a blood sample at least once through the first opening of the device and passing it through the membrane filter to separate plasma from the blood sample; And

c) after a predetermined time has elapsed, collecting the separated plasma in a plasma chamber by applying a negative pressure by means of a negative pressure applying means connected to the second opening;

There is provided a method of separating plasma from a blood sample comprising a.

According to an exemplary embodiment, the negative pressure applying means may be a syringe pump or a peristaltic pump.

The device for separating plasma from blood, provided in embodiments of the present disclosure, can be embodied in a simple structure that basically includes an upper structure and a lower structure, and further, a plasma separation device can be manufactured through a simple assembly operation. . In addition, by using the capillary phenomenon during the separation process of the plasma in the membrane filter and / or the movement of the separated plasma has the advantage that the separation and collection efficiency of the plasma can be improved.

In particular, since the plasma separation device is configured to collect plasma in the plasma chamber only by applying a negative pressure to the second opening, and has a structural characteristic suitable for constructing a plurality of modules, the plasma separation device may have a system for collecting blood (or whole blood) from a shear or By connecting the module and the diagnostic system or the module of the plasma collected at the rear end, it is possible to implement a series of automated immunoassay systems consisting of the collection of blood, the separate recovery of plasma from the blood and the diagnosis using the collected plasma in an automated manner.

In addition, since the main constituent members of the device can be produced by injection molding, the device is easy to manufacture and is suitable for mass production. Therefore, broad commercialization is expected in the future.

1 is a perspective view showing an appearance of an assembled state of a device for separating plasma from blood according to one embodiment;
2 is an exploded perspective view of a device and membrane filter in combination with a device for separating plasma from blood according to one embodiment;
3 is a diagram illustrating an inner surface structure of an upper structure of a device for separating plasma from blood according to one embodiment;
4 is a top view of an undercarriage of a device for separating plasma from blood according to one embodiment;
5A-5C are, in one embodiment, a longitudinal cross-sectional view showing, in one embodiment, a state in which a membrane filter is seated in an underlying structure of a device for separating plasma from blood, and (b) a membrane filter and a second structure in the underlying structure; A longitudinal cross-sectional view with the first and second sheets combined, and (c) a longitudinal cross-sectional view with the membrane filter and the first and second sheets combined (assembled) in a device for separating plasma from blood;
FIG. 6 shows the principle of operation of connecting the negative pressure applying means in an automated manner by an external drive to a device for separating plasma from blood according to an exemplary embodiment;
7 is an exploded perspective view of a device for separating plasma from blood according to another embodiment;
8 is a longitudinal sectional view with the device assembled for separating plasma from blood according to another embodiment assembled; And
9A to 9D are photographs showing a series of processes for separating plasma from blood using the device fabricated in Example 1; And
10A to 10C are photographs showing a series of processes for separating plasma from blood using the device manufactured in Example 2, respectively.

The present invention can all be achieved by the following description. The following description is to be understood as describing preferred embodiments of the invention, but the invention is not necessarily limited thereto. In addition, the accompanying drawings are for ease of understanding, the present invention is not limited thereto, and the details of individual configurations may be appropriately understood by the specific gist of the related description to be described later.

The terminology used herein may be understood as follows.

"Blood" can generally mean whole blood, and in some cases can also include additional ingredients such as saline, nutrients and / or anticoagulants. It may also include blood that has been pretreated to remove some components, such as some of the white blood cells, from the whole blood.

"Path" or "channel" means a path through which a fluid (especially a liquid fluid) moves in a predetermined direction, It is not necessarily limited to the closed form, it can be understood as a concept including the open form.

A “fine pattern” has at least one cross-sectional dimension (eg, width, depth, height, diameter, etc.) of the protrusions or recesses of the structure constituting the pattern, for example, about 10 mm or less, specifically about 5 mm. Hereinafter, more specifically, the pattern may be about 1 mm or less.

The expressions "on" and "on" may be understood to be used to refer to relative positional concepts. Thus, not only when another component or layer is directly present in the layer mentioned, there may be at least one other layer (intermediate layer or intervening layer) in between, or an additional component may be present or present. Similarly, the expressions "below", "below" and "below" and "between" may also be understood as relative concepts of position. The expression "sequentially" may also be understood as a relative positional concept.

The term "contacts" means, in consultation, direct contact between two materials, but in broad terms it may be understood that any additional component may be interposed, as long as contact between the component and the liquid flow is made. .

If any component or member is described as being “connected” or “communicated” with another component or member, unless otherwise stated, as well as directly connected or in communication with said other component or member, as well as It is to be understood that this also includes the case where they are connected or communicated under the intervening of other components or members.

According to the present disclosure, a chip-type device for separating plasma from blood may largely include an upper structure and a lower structure. In the case of the superstructure, a blood (specifically whole blood) sample can be injected into the device from the outside and serve as a connection with negative pressure applying means for transporting the plasma separated by the membrane filter. In addition, plasma may be efficiently discharged downwardly of the membrane filter by supporting or pressing a membrane filter seated (or mounted) in the device by using a protruding member formed on the surface of the upper structure (inner space direction of the device) if necessary. To increase the capillary phenomenon. On the other hand, the lower structure corresponds to a structure that separates the plasma from the blood injected through the membrane filter, by moving it to receive and store the separated plasma in a predetermined space, that is, the plasma chamber.

In particular, it is noted that both the upper structure and the lower structure can be manufactured as a polymer material in an integrated form by a molding process (specifically, an injection molding mold process), and thus manufacturing is easy, and manufacturing cost can be significantly lowered.

First embodiment

FIG. 1 shows an appearance of an assembled state of a device for separating plasma from blood according to one embodiment, and FIG. 2 is an exploded perspective view of a device and a membrane filter combined in the above embodiment.

Referring to the drawings, the upper structure 101 and the lower structure 102 constituting the device 100 for separating the plasma from the blood are each manufactured in a predetermined shape, it can be assembled by mounting the membrane filter 111 have. At this time, the material of each of the upper structure 101 and the lower structure 102 may be the same or different from each other as a polymer material. In addition, even if the same or homogeneous polymer material, the individual physical properties of each of the upper structure and the lower structure may be different in some cases.

As one example, the polymer having a suitable property for the injection molding process is preferred, for example, may be a polymer having good flowability under melt conditions during injection. According to certain embodiments, the polymer has a melt index as measured by ASTM 1238, for example in the range of about 0.1 to 100 g / 10 min, specifically about 4 to 60 g / 10 min, more specifically 10 to 50 g / 10 min. It may be, but is not necessarily limited thereto.

In addition, the polymer forming the upper structure 101 and the lower structure 102 may be a material having transparency, specifically transparency. Transparency does not have a substantial effect on the performance of the device, but transparency is preferably possible to visually determine the extent to which the injected blood diffuses on the membrane filter 111 and / or the amount of plasma collected in the plasma chamber 126. It may be advantageous to use this high polymer material.

Examples of the polymer material constituting the upper structure 101 and the lower structure 102 include polyester (specifically, polyethylene terephthalate (PET)), polyethylene (PE), polypropylene (PP), polyimide (PI), Polystyrene (PS), polycarbonate (PC), polyurethane, polyvinylidene fluoride, nylon, polymethyl methacrylate (PMMA), polyvinyl chloride (PVC), cyclic olefin copolymer (COC), liquid crystal polymer ( LCP), polyamide (PA), polyimide (PI), poly (phenylene ether) (PPE), polyoxymethylene (POM), polyether ether ketone (PEEK), polyether sulfone (PES), polytetrafluoro Low ethylene (PTFE), an acrylic polymer, a silicone-based polymer, and the like. In an exemplary embodiment, the polymer to be molded may be an elastomer having elasticity, specifically a silicone-based polymer, more specifically polydimethyl siloxane (PDMS or h-PDMS), polymethylsiloxane, partially alkylated polymethylsiloxane, Polyalkylmethylsiloxanes, polyphenylmethylsiloxanes, combinations thereof, and the like. In particular, PDMS can be used.

In the illustrated embodiment, the upper structure 101 and the lower structure 102 may be bonded to each other (or bonded) in such a way that the adhesive is interposed. Such an adhesive is not particularly limited as long as it has sufficient adhesiveness to mutually bond each layer constituting the microfluidic device from blood to form a flow path. For example, a rubber adhesive, an acrylic resin adhesive, a silicone adhesive, an optical adhesive, a heat adhesive, or the like can be used. In this case, the adhesive may be specifically applied in the form of a single-sided or double-sided adhesive tape, more specifically in the form of a double-sided adhesive tape, for example, a pressure-sensitive adhesive tape, a thermally active adhesive tape, a chemically active adhesive tape, a photoactive adhesive tape, and the like. have.

On the other hand, according to the illustrated embodiment, it has a pentagonal shape (or a shape in which both corner portions of one side are cut in a rectangle).

On the other hand, referring to Figure 2, the membrane filter 111 for separating only the plasma components from the blood (specifically whole blood) is a blood chamber formed after the assembly of the upper structure 101 and the lower structure 102, specifically described later As shown in FIG. 2, the membrane filter seat 123 may be mounted in the lower structure 102. In the illustrated example, the membrane filter 111 may be manufactured to have a shape that matches the shape of the membrane filter seat 123.

As the membrane filter 111, blood introduced from the upper side (especially whole blood) can easily penetrate into the membrane, quickly and efficiently discharge the plasma separated below the membrane, and separate the separated plasma into a plasma chamber ( 126) in the process of applying a negative pressure is advantageous to have a property that can separate only the plasma while inhibiting the hemolysis, such as red blood cells in the blood passing through the membrane as much as possible.

In this regard, according to an exemplary embodiment, the thickness of the membrane filter 111 may be in the range of, for example, about 100 to 1,000 μm, specifically about 200 to 500 μm, more specifically about 300 to 350 μm. If the thickness of the membrane is too thin or too thick, red blood cells may be hemolyzed or the filter may break down due to low negative pressure during plasma separation, which may cause whole blood leakage or increase the amount of plasma that the filter itself contains and is not recovered. In consideration of the amount of blood to be injected, etc. can be appropriately adjusted within the above-mentioned range.

According to an exemplary embodiment, the membrane filter 111 comprises at least one porous medium, and the structure that can be discharged by filtering in the downward direction of the membrane by selectively moving only the plasma in the blood in the thickness direction through the membrane and It may be made of a material. In this regard, the size of the pores in the membrane is, for example, about 0.1 to 100 μm, specifically about 0.5 to 50 μm, more specifically about 1, taking into account the size of blood cells in the blood (about 2. to 10 μm). To 20 μm. For example, blood cells in the blood can have a pore size range that prevents them from passing through the membrane.

According to one embodiment, the porous membrane may be, for example, an isotropic membrane, an anisotropic membrane or a combination thereof. In this regard, as anisotropic membranes, asymmetric membranes of the same material and composite membranes of two or more different materials can be exemplified. According to certain embodiments, anisotropic membranes, in particular asymmetric membranes, may be used, for example, such that the upper region of the membrane has a relatively large pore size, while the lower region has a relatively small pore size, based on thickness. Can be configured. By way of example, the pore size of the upper region in the membrane may range from, for example, about 5 to 100 μm, specifically about 10 to 50 μm, more specifically about 20 to 40 μm, while the pore of the lower area in the membrane The size may range from, for example, about 0.1 to 5 μm, specifically about 0.5 to 2 μm, more specifically about 0.5 to 1 μm. In this regard, one may exhibit a pattern in which the pore size continuously changes (eg, continuously decreases pore size) as it progresses from the top surface to the bottom surface of the membrane.

In this way, an asymmetric membrane is used, but by reducing the pore size in the flow direction of the fluid during filtration, it is not only a fluid movement by absorption, but also induces a capillary phenomenon, so that the fluid, in particular, the separated plasma is directed downward of the membrane. It can be transported easily and quickly.

According to an exemplary embodiment, the material of the membrane may be a polymer, for example, polyolefin, polyester, polyamide, polysulfone, acrylic polymer, polyacrylonitrile, polyaramid, polymer of halogenated olefin, combinations thereof, and the like. Can be. Specifically, polyethylene, polypropylene, polyethylene terephthalate (PET), nylon (eg, nylon 6, 66, etc.), polyvinylidene difluoride (PVDF) can be exemplified.

According to an alternative embodiment, for example, cellulose derivatives, specifically cellulose acetate, cellulose propionate, cellulose acetate propionate, cellulose acetate butyrate, and the like may be used as the natural polymer system.

Meanwhile, in the case of the upper structure 101, at least one first opening 103 formed to first inject or inject a blood sample is formed. In the illustrated embodiment, one end region (not shown) of the upper structure 101 is illustrated. In this example, the area is near the corner of the pentagon. In this case, the first opening 103 may be formed of a cavity having a variety of shapes (for example, circular, elliptical, etc., more specifically, circular). In particular, when the circular cavity is formed, the cavity of the quadrangle during blood injection is formed. The phenomenon which overflows to the outside compared with etc. can be suppressed. In addition, the cavity constituting the first opening may be preferably sized to inject a blood sample into the blood chamber in the device 100 using a pipette or the like. By way of example, the size (or diameter) of the first opening 103 may range, for example, from about 1 to 100 mm, specifically about 5 to 50 mm, more specifically about 8 to 20 mm.

Meanwhile, in the illustrated embodiment, the air outlet 104 may be further included together with the cavity forming the first opening 103. More specifically, the first opening and the air outlet may be formed in a fused form. have. The air outlet 104 allows the air pushed out by the blood to be easily discharged to the outside as the blood sample is injected, thereby providing the advantage that the blood sample is quickly introduced into the space, that is, the blood chamber, without air resistance. have. According to an exemplary embodiment, the first opening 103 may have a shape in which air outlets are fused in a wing shape or an ellipse shape on both sides of the circular cavity.

Meanwhile, in the illustrated upper structure 101, a second opening portion 105 configured to be connected to a negative pressure applying means for recovering plasma is formed in an end side region in a direction opposite to the first opening portion 103. The second opening 105 is in communication with the plasma chamber 126 provided in the lower structure 102, through which the negative pressure applying means (for example, a syringe pump) is connected to the negative pressure applied inside the device 100 Thereby moving the plasma separated from the blood to provide a driving force that can be collected in the plasma chamber 126.

According to an exemplary embodiment, the upper structure 101 may be a flat plate-based structure, the thickness of which is for example about 0.5 to 20 mm, specifically about 1 to 15 mm, more specifically about 5 to about 10 mm range, but the present invention is not limited thereto.

3 shows the inner structure of the upper structure of the device for separating plasma from blood according to one embodiment.

Referring to the drawings, a plurality of protruding members 106 are formed on the rear surface of the upper structure 101 (that is, the surface facing the internal space of the device) at regular intervals. The protruding member 106 may be integrally formed when the upper structure is manufactured by molding (injection molding).

In the present embodiment, the shape and dimensions of the protruding member 106 are not particularly limited, but the lower structure 102 on which the upper structure 101 and the membrane filter 111 are seated or mounted as shown in FIG. 2. By pressing the surface of the membrane filter 111 seated in the lower structure 102 during the assembly of the fluid (ie, separated plasma) by the capillary phenomenon in a fine pattern on the seating portion 123 of the membrane filter as described below It can provide an effect of enhancing movement.

According to the exemplary embodiment, the number and spacing of the protruding members 106 formed in the upper structure 101 are also not particularly limited as long as the membrane filter 111 can be sufficiently pressed against the lower side. However, the spacing between the plurality of protruding members 106 is, for example, about 5 to 50 mm, specifically about 7 to about the longitudinal direction of the upper structure 101 (ie, the direction of the first opening to the second opening). 20 mm, more specifically in the range of about 8 to 10 mm. In particular, when the material of the protruding member 106 is made of a material having elasticity, damage to the membrane filter 111 in contact with the lower side can be suppressed, and the applied pressure can be distributed relatively uniformly.

4 is a plan view of an undercarriage of a device for separating plasma from blood, respectively, according to one embodiment, and each of FIGS. 5A-5C is a substructure of a device for separating plasma from blood, in one embodiment. A longitudinal cross-sectional view showing a state in which a membrane filter is seated on the membrane, (b) a longitudinal cross-sectional view of a membrane filter and a first sheet and a second sheet in a lower structure, and (c) a device for separating plasma from blood. It is a longitudinal cross-sectional view of the state in which the membrane filter and the 1st and 2nd sheet were combined.

Referring to the drawings, the lower structure 102 as a whole forms the appearance skeleton by the frame member 121. In the frame member 121, an edge member 122 having a predetermined width inwardly is formed along its outer circumferential surface to partition an area for mounting or mounting the membrane filter 111, and in some cases, the upper structure 101. ) May serve to support the upper structure when assembled. Accordingly, when the upper structure 101 and the lower structure 102 are assembled, a blood chamber may be formed to receive the blood injected into the space between the upper structure and the lower structure. Alternatively, the edge member described later may be omitted, and as described above, the plurality of protruding members 106 formed on the upper structure 101 may be connected to the membrane filter seat 123 through the membrane filter 111. It may also serve as a support structure for forming a blood space upon contact.

According to the embodiment shown, the membrane filter seat 123 is tilted symmetrically or asymmetrically (more specifically symmetrically) downward from both sides of the frame member 121 of the lower structure 102. And, as a result, a pair of inclined surfaces are provided on the membrane filter seat 123 (see FIG. 4). At this time, the inclination angle (based on the horizontal line) forming the inclined surface from both sides of the frame member 121 is, for example, about 5 to 80 °, specifically about 10 to 60 °, more specifically about 20 to 40 ° range It can be determined in consideration of the wettability and flowability of blood on the membrane filter within.

According to the above embodiment, the pair of inclined surfaces formed while being inclined downwardly from both sides of the frame member 121 in the lower structure do not meet each other, but are spaced apart at predetermined intervals from the central region. The area bounded by the spaced apart in this way extends along the length direction of the membrane filter seat 123 and also serves to function as a passage while forming a substantially flat surface. Plasma migration pathway ". As such, the liquid plasma (plasma separated from the blood by the membrane filter) flows from each of the pair of inclined surfaces by the inclined structure formed from both sides of the lower structure 102, and thus, the first plasma movement passage ( 124). At this time, the width of the first plasma passage 124 is, for example, about 0.1 to 5 mm, specifically about 0.5 to 3 mm, more specifically about the volume of the separated plasma and the intensity of the applied negative pressure. It may range from 0.8 to 1 mm, but the present invention is not limited thereto.

In the illustrated embodiment, the separated plasma flows through the first plasma passage 124 by the application of negative pressure as described below, wherein the first plasma passage 124 extends while inclined downward. Because of this, plasma flows in the direction of the second opening 105 more easily and quickly by the action of gravity.

By way of example, the inclination angle (horizontal basis) at which the first plasma movement passage 124 is inclined is, for example, within a range of about 0.5 to 30 °, specifically about 2 to 10 °, and more specifically about 3 to 5 °. It can be determined in consideration of the wettability and flowability of the separated plasma. As such, when the first plasma movement passage 124 is configured to move along the inclined surface in the direction of the second opening, the membrane filter seat 123 is also inclined in the longitudinal direction (that is, the inclined surface on which the membrane filter is seated). Formed), the injected blood can soak the membrane filter as much as possible. Thus, injecting a relatively small amount (eg, about 1 mL) of a blood sample provides the advantage of ensuring the amount of plasma required for diagnosis or analysis.

Meanwhile, according to the illustrated embodiment, a pair of inclined surfaces symmetrically or asymmetrically (i.e., in the process of converging the plasma separated from the blood injected through the membrane filter 111 into the first plasma movement passage 124) , A pair of surfaces inclined symmetrically or asymmetrically) is subjected to the first guide protrusion pattern B and the second guide protrusion pattern B '.

In an exemplary embodiment, each of the first guide protrusion pattern B and the second guide pattern B ′ may be a symmetrical or asymmetrical pattern, and a fixed or atypical pattern. In addition, each of the first guide protrusion pattern B and the second guide protrusion pattern B ′ is selected from the group consisting of a comb-tooth pattern, a vertical line pattern, a horizontal line pattern, a wave pattern, a dot pattern, a wrinkle pattern, and a sash pattern. It may be at least one protruding pattern. Therefore, in the illustrated example, although the protrusion pattern in the form of a comb pattern is formed, it may be understood that the present invention is not limited thereto.

In this regard, since the plasma separated from the membrane filter 111 and discharged or dropped to the lower side contains a large amount of water, wettability is low on the inclined surface of the hydrophobic polymer material, and consequently converges effectively to the first plasma passage 124. It can be difficult to do. However, the insufficient fluid movement phenomenon can be alleviated by forming a plurality of protrusions constituting the guide protrusion patterns B and B 'on each of the inclined surfaces. In this case, the separation distance between the plurality of protrusions constituting the guide pattern functions as a fluid channel, and the upper surface of the protrusion may induce capillary phenomenon in contact with the lower surface of the membrane filter 1411 wetted by the injected blood. have.

In the illustrated embodiment, the protrusions of the guide pattern B extend from the both sides of the membrane filter seat 123 toward the first plasma passage 124, with the membrane filter seat 123. Arranged in a shape inclined in the direction of the second opening on a pair of symmetrical or asymmetric inclined surfaces formed on the substrate. In the example shown in FIG. 4, the guide protrusion patterns B and B 'in the form of comb-shaped patterns are arranged in a wedge shape as a whole while forming symmetry with each other.

According to certain embodiments, when the guide patterns B and B 'are comb patterned, the spacing between the plurality of protrusions constituting them is about 0.5 to 1 mm, specifically about 0.6 to 0.9 mm, more specifically about 0.7 To 0.8 mm. In this regard, it may be difficult for the fluid to move easily by the capillary phenomenon in the above-described numerical range. Therefore, the height of the protrusion can be formed to be relatively low to induce capillary phenomenon, for example, to adjust within the range of about 0.05 to 0.5 mm, specifically about 0.07 to 0.3 mm, more specifically about 0.1 to 0.2 mm Can be.

Furthermore, as described above, the plurality of protruding members 106 spaced apart from the lower surface of the upper structure 101 are seated on the membrane filter seat 123 when the upper structure 101 and the lower structure 102 are assembled. Capillary force may be further increased by pressing the membrane filter 111 to closely contact the protrusions of the guide patterns B and B '.

As such, the plasma rapidly transported downward along the pair of symmetrical inclined surfaces by the guide patterns B and B 'is framed through a hole formed at or near the downstream end region of the first plasma movement passage 124. It may be connected to the second plasma movement path 125 formed across the bottom thickness of the member 121. According to an exemplary embodiment, the diameter of the second plasma migration passage 125 may range from, for example, about 0.1 to 5 mm, specifically about 0.5 to 3 mm, more specifically about 1 to 2 mm, The dimension may be changed in consideration of the amount of blood injected into the device, the amount of separated plasma, and the like.

4 and 5, in the frame member 121 of the lower structure 102, a plasma chamber 126 corresponding to and in communication with the second opening 105 of the upper structure 101 is provided. have. The receiving volume of fluid in the plasma chamber 126 may, for example, range from about 0.01 to 1 mL, specifically about 0.1 to 0.5 mL, and the size (diameter) of the upper surface of the plasma chamber may be, for example, from about 2 to 20 mm, specifically about 5 to 15 mm, more specifically about 7 to 10 mm range. However, the numerical range may be understood in an exemplary sense.

In the illustrated embodiment, the first plasma movement passage 124 extends in an inclined manner, but the plasma chamber 126 is substantially perpendicular to the bottom surface of the frame member 121. Therefore, even if it is desired to transfer the plasma collected by the micropipette after the plasma collection, the tip of the pipette can still be introduced into the plasma chamber 126 in the vertical direction, so that the first plasma movement passage 124 is easily inclined. Plasma may be recovered or collected.

In addition, the bottom surface of the frame member 121 is attached to the first sheet 112 and the lower surface of the first sheet 112 and the cavity having a shape extending in the longitudinal direction at the portion overlapping with the second plasma movement passage 125 or The bonded second sheet 114 is attached to the bottom surface of the lower structure 102 to form the plasma moving channel 113. The cavity may be formed in a predetermined or non-uniform distance in the inner direction along the outer periphery of the first sheet 112. In the illustrated example, the cavity is formed in a straight pattern, but in some cases, a curved pattern ( For example, spiral, serpentine, zigzag, etc.). In addition, the cavity forming method is not particularly limited, but a laser cutting, a cutting plotter processing method, a cutting printing method, a lathe processing method, or the like may be applied, and each of these technical principles and detailed technical details is known in the art. Also, for example, the width of the plasma moving channel 113 may be, for example, in the range of about 100 to 2,000 μm, specifically about 200 to 1,500 μm, more specifically about 500 to 1,000 μm, which is illustrative. It can be understood as.

In addition, an adhesive may be interposed between the bonding of the first sheet 112 and the second sheet 114, or the combination of the first and second sheets to the bottom surface of the frame member 121. It is not limited to a specific kind as long as it has sufficient adhesiveness to form a flow path. For example, a rubber adhesive, an acrylic resin adhesive, a silicone adhesive, an optical adhesive, a heat adhesive, or the like can be used. In this case, the adhesive may be specifically applied in the form of a single-sided or double-sided adhesive tape, more specifically in the form of a double-sided adhesive tape, for example, a pressure-sensitive adhesive tape, a thermally active adhesive tape, a chemically active adhesive tape, a photoactive adhesive tape, and the like. have. However, as long as the components of the adhesive may contaminate the plasma when exposed to the plasma transfer channel 113, it is desirable to prevent the adhesive from contacting the moving plasma as much as possible.

In addition, the thickness of each of the first sheet 112 and the second sheet 114 may be different or the same, for example, about 20 μm to 1 mm, specifically about 20 to 300 μm, more specifically about 50 to It may be appropriately selected within the range of 100 μm. In particular, the thickness of the first sheet 112 will substantially determine the height of the plasma migration channel. However, when the adhesive is used, the height of the adhesive layer also affects the height of the plasma transfer channel, but the effect is slightly less than the thickness of the first sheet. As such, the plasma transfer channel formed by the two sheets 112 and 114 may be manufactured in the form of an adhesive film, and the device may be completed by attaching it to a predetermined portion of the bottom surface of the lower structure 102.

As such, the plasma movement channel 113 formed by the two sheets 112 and 114 may include the second plasma movement passage 125 and the plasma chamber 126 together with the communication tube 127 formed at the lower end of the plasma chamber 126. Are communicated with each other. In this case, the diameter of the communication tube 127 may be, for example, about 0.2 to 5 mm, specifically about 0.5 to 2 mm, but this may be changed. In the illustrated embodiment, the cavity 113 can be configured in a manner that overlaps the lower end of the communication tube 127 as well as the second plasma migration passage 125 as described above to form a flow path for the fluid. .

In an exemplary embodiment, the first sheet 112 and the second sheet 114 may be composed of a polymer film. Examples of the polymer constituting such a film-like sheet include polyester (specifically, polyethylene terephthalate (PET)), polyethylene (PE), polypropylene (PP), polyimide (PI), polystyrene (PS), polycarbonate (PC), polyurethane, polyvinylidene fluoride, nylon, polymethyl methacrylate (PMMA), polyvinylchloride (PVC), cyclic olefin copolymer (COC), liquid crystal polymer (LCP), polyamide (PA ), Polyimide (PI), poly (phenylene ether) (PPE), polyoxymethylene (POM), polyether ether ketone (PEEK), polyether sulfone (PES), polytetrafluoroethylene (PTFE), acrylic It may be a resin of the system. More specifically, it may be polyethylene terephthalate (PET), but is not necessarily limited thereto. In addition, the first and second sheets 112 and 114 may be light-transmitting materials, in particular, a film-like sheet made of a transparent polymer material. Whether the separated plasma is smoothly transferred to the plasma chamber 126 upon application of negative pressure The back can be observed with the naked eye.

 According to the illustrated embodiment, when the negative pressure is applied by the negative pressure applying means connected through the second opening 105, the plasma separated by the membrane filter 111 is the first plasma passage 124, the second plasma passage ( 125, and through plasma moving channel 113 and communication tube 127, are collected in plasma chamber 126.

On the other hand, the blood chamber (especially the membrane filter seat in the underlying structure), the first plasma passage and the connecting means between the first plasma passage and the plasma chamber (in the example shown, the second plasma passage 125, the plasma The moving channel 113 and the communicating tube 127 are both polymeric materials and their surfaces show hydrophobicity (or nonpolarity) or weak hydrophilicity. Accordingly, at least one surface of the polymer material may be hydrophilized to improve plasma flowability. Specifically, the hydrophilic surface is a surface that attracts moisture, and blood or plasma is spread on the hydrophilic surface. Water drops on hydrophilic surfaces have the property of having low water contact angles at the interface, while hydrophobic surfaces exhibit high water contact angles. For this reason, in the case of hydrophobic or low hydrophilic surfaces, the flow characteristics of blood or plasma may not be good, which may reduce the flow rate of the fluid and thereby lower the plasma collection efficiency. In this regard, it may be advantageous to improve the flow characteristics of the fluid, for example by exhibiting a water contact angle of about 60 ° or less, specifically about 40 ° or less, more specifically about 20 ° or less.

For this purpose, organic and / or inorganic materials can be used to hydrophilize, which would be advantageous to use surface treatment materials that do not substantially react (ie, are inactive) to blood and separated plasma in contact with the surface in the device. Can be. According to an exemplary embodiment, the organic surface treatment may include, for example, a compound having an amine, hydroxy, carbonyl, or epoxy functional group, a monomer having the functional group, a dimer, and Polymers and the like, and these may be used alone or in combination of two or more. In addition, the inorganic surface treatment product is, for example, a metal or a nonmetal, specifically, at least one metal selected from gold, silver, silicon, aluminum, nickel, iron, copper, manganese, silicon, titanium, chromium, or the like, or a combination thereof. It may be an oxide. In some cases, the organic surface treatment and the inorganic surface treatment may be combined, and the organic and inorganic surface treatment agents may be mixed, or any one of the organic surface treatment and the inorganic surface treatment may be performed first, followed by the remaining surface treatment.

Organics and / or inorganics can be sugars such as thermal evaporation, E-beam evaporation, sputtering, chemical vapor deposition (CVD), sol-gel, plasma treatment, liquid phase chemical treatment, gas phase chemical treatment, etc. It may be formed by a coating or attachment method known in the art. According to an exemplary embodiment, the thickness of the organic and / or inorganic coating layer formed on the surface of the polymer material may be selected, for example, within the range of about 1 to 500 nm, specifically about 1 to 200 nm. It may be understood in an exemplary manner. Alternatively, it may be treated with a biological material such as BSA (Bovine serum albumin).

In the present embodiment, the negative pressure applying means connected to the second opening 105 to collect the separated plasma is not particularly limited, but a pump known in the art, for example, a syringe pump, a peristaltic pump, or the like is applied. can do. In this case, the applied negative pressure may be, for example, about 0.1 to 1 kg / cm 2, specifically about 0.2 to 0.9 kg / cm 2, and more specifically about 0.5 to 0.8 kg / cm 2. If the applied negative pressure is too high, hemolysis may occur as erythrocytes in the blood pass through the membrane filter 111, while under an insufficiently low negative pressure condition, as much as it can reduce the transport efficiency of the separated plasma. It may be desirable to adjust appropriately within the aforementioned range. As such, when the negative pressure is optimized, unhemolytic yellow plasma can be collected in the plasma chamber.

According to an exemplary embodiment, after injecting a blood sample into the device, the blood pressure is allowed to penetrate the membrane filter over a period of time (eg, about 0.5 to 5 minutes, specifically about 1 to 2 minutes), followed by Can be authorized. In addition, the negative pressure application time may vary depending on the amount of the blood sample injected, for example, may be in the range of about 1 to 10 minutes, specifically about 2 to 5 minutes.

Subsequently, when a sufficient amount of plasma is collected, the second opening 105 of the upper structure 101 is disconnected from the negative pressure applying means, and at least a part of the plasma collected by the micropipette (not shown) is diagnosed. Can be collected for

According to an exemplary embodiment, connecting the negative pressure applying means to the plasma separation device 100 can be implemented in an automated manner, an exemplary embodiment of which is shown in FIG. 6.

Referring to the drawings, a funnel or longitudinal coupling member 131 is fixed to one end region of the support member 132. In the illustrated example, the support member 132 is configured as a rod, but as long as it can support the coupling member 131, various modifications are possible. In addition, the other end of the support member 132 is mechanically connected to the rotation member 133 and configured to repeat the semi-rotational movement by the operation of the rotation member 133. At this time, the rotation member 133 is connected to the motor 134 and the conveyor belt, for example, to provide the rotation driving force to the support member 132.

On the other hand, the plasma separation device 100 is mounted in a fixed plate 135, specifically, a concave or groove region formed in the fixed plate 135 and fixed to a predetermined position, and the coupling member ( 131 is in close contact with the second opening 105. To this end, the coupling member 131 is provided with a suction device (not shown) therein, the material having elasticity, such as an elastic rubber material, such as when in contact with the second opening 105 in a sealed or sealed state The outer shell structure can be formed.

If the plurality of plasma separation devices are mounted (fixed) in the structure in which the plurality of fixed plates are formed, the negative pressure applying means may also be provided to match the number of devices to simultaneously separate the plasma.

As described above, the plasma separation device 100 according to the present embodiment can be implemented by simply assembling the main structure of the upper structure 101 and the lower structure 102 in the form of an integral molding made of a polymer material. In addition, the same or different blood samples may be injected simultaneously into each of the plurality of devices by automated injection means, and plasma may be collected within a short time by applying negative pressure at one time. Collected plasma components can also be collected by an automated method such as a micropipette to diagnose various diseases using a diagnostic chip or kit. In this case, from the blood sample to the diagnosis through a system including a separation and collection module of plasma from blood, a diagnosis module for disease using the collected plasma, a blood sample and plasma transfer module (syringe module), a pipette tip replacement module, and the like. A series of steps can be performed in a modular and automated manner.

Second embodiment

7 is an exploded perspective view of a device for separating plasma from blood according to another embodiment of the present disclosure, and FIG. 8 is a longitudinal cross-sectional view of the device in an assembled state.

In the description of this embodiment, members corresponding to the members described in the first embodiment will be described using similar member numbers. For example, the upper structure 101 in the first embodiment is specified as the upper structure 201 in this embodiment. Therefore, below, only technical matters distinguished from the 1st specific example are described, and common technical matters are abbreviate | omitted.

In the illustrated embodiment, a pair of first openings 203 having a shape extending along the longitudinal direction of the upper structure 201 is formed. Thus, unlike the first embodiment described above, the blood sample may be divided into each of the first openings, and also divided into each of the first openings so as to penetrate the membrane filter portion 211 more quickly. It can also be injected. In addition, in the illustrated embodiment, a separate air outlet may not be required because of the low possibility of air thrust caused by blood.

In the case of a device according to the embodiment shown, it may be advantageous to process a relatively large amount of blood sample to obtain plasma. For example, in the case of the device 100 according to the first embodiment, it may be suitable for separating plasma from a small amount (eg, about 0.5 to 1.5 mL) of a blood sample, while the device according to the present embodiment ( 200) can process larger amounts (eg, about 1.5-2.5 mL) of blood samples. In addition, after infusion of the blood sample, it is allowed to soak over a predetermined time (for example about 0.5 to 5 minutes, specifically about 1 to 2 minutes), and then the negative pressure, for example about 1 to 10 minutes, specifically about It may be applied over 2 to 5 minutes.

Further, according to the illustrated embodiment, the second opening 202 or the plasma chamber (not being inclined) is not inclined by the first plasma movement passage 224 bounded by two symmetrical inclined surfaces of the membrane filter seat 223. 226 direction. Thus, unlike in the first embodiment, the top surface of the frame member 221 in the lower structure 202 is configured to be substantially horizontal.

Example 1

The plasma separation device according to the first embodiment was manufactured by itself, and an external planar photograph thereof is shown in FIG. 9A.

A. Details regarding the plasma separation device used are as follows.

(i) superstructure 101

-Material: Acrylic resin (PMMA)

Thickness: 1 mm

Width and length: 30 mm and 71 mm respectively

Height and spacing of the protruding member 106: 5 mm and 11.5 mm, respectively.

Diameter of the first opening 103: 7 mm

(ii) substructure 102

-Material: Acrylic resin (PMMA)

Maximum height and minimum height of the frame member 102: 15 mm and 10 mm, respectively

Width of the rim member 122: 2.5 mm

The angle of inclination of the pair of surfaces symmetrically inclined from both sides of the undercarriage of the membrane filter seat: 30 °

Width of the first plasma passage 124: 0.7 mm

Fluid flow direction tilt angle of the first plasma passage 124: 4 °

-Height and spacing of comb pattern: 0.2 mm and 0.7 mm, respectively

Diameter of the second plasma migration passage 125: 1.5 mm

(iii) Membrane Filters: Pall's Vivid Membrane Filters

(iv) plasma transfer channels

Material of the first and second sheets 112, 114: PVC and PET, respectively

Thicknesses of the first and second sheets 112, 114: 230 μm and 100 μm, respectively

Width and length of cavity 113: 1.5 mm and 11.8 mm, respectively.

B. Plasma Separation Experiments from Blood Samples

Blood sample (1 mL) was maintained for about 2 minutes after injection through the first opening. As a result, blood penetrated the membrane filter as shown in Fig. 9B.

Thereafter, a negative pressure of 0.7 kg / cm 2 was applied over 4 minutes using a cylinder pump connected to the second opening 105 as illustrated in FIG. 9C, and the membrane filter 111 was introduced into the plasma chamber 126. It was confirmed that the separated plasma was collected.

As a result of plasma collection through negative pressure application, 170 μl of plasma was collected from the plasma chamber using a micropipette, which is shown in FIG. 9d. According to the figure, it was confirmed that only plasma components can be effectively recovered and recovered from blood (whole blood) within a short time without hemolysis.

Example 2

The plasma separation device according to the second embodiment was manufactured by itself, and an external photograph thereof is shown in FIG. 10A.

A. Details regarding the plasma separation device used are as follows.

(i) superstructure 201

-Material: Acrylic resin (PMMA)

Thickness: 1.mm

Width and length: 30 mm and 71 mm respectively

Height and spacing of the protruding member 206: 5 mm and 11.5 mm, respectively.

Longest length and width of the first opening 203: 46.5 mm and 6 mm, respectively.

(ii) substructure 202

-Material: Acrylic resin (PMMA)

Height of the frame member 202: 10 mm

Width of the rim member 222: 2.5 mm

The angle of inclination of the pair of surfaces symmetrically inclined from both sides of the undercarriage of the membrane filter seat: 30 °

Width of the first plasma passage 224: 0.7 mm

-Height and spacing of comb pattern: 0.2 mm and 0.7 mm, respectively

Diameter of the second plasma passage 225: 1.5 mm

(iii) Membrane Filters: Pall's Vivid Membrane Filters

(iv) plasma transfer channels

Material of the first and second sheets 212, 214: PVC and PET

Thicknesses of the first and second sheets 212, 214: 230 μm and 100 μm, respectively

Width and length of cavity 213: 1.5 mm and 11.8 mm, respectively

B. Plasma Separation Experiments from Blood Samples

Blood samples (2 mL) were divided into two injections into each of the pair of first openings and maintained for about 2 minutes, resulting in blood seeping into the membrane filter.

Then, as shown in FIG. 10B, a negative pressure of 0.7 kg / cm 2 was applied over 3 minutes and 30 seconds using a cylinder pump connected to the second opening 205, and the membrane filter 211 in the plasma chamber 226. It was confirmed that plasma separated by was collected.

As a result of plasma collection through negative pressure application, 154 μl of plasma was collected from the plasma chamber using a micropipette, which is shown in FIG. 10c. According to the figure, it was confirmed that only plasma components can be effectively recovered and recovered from blood (whole blood) within a short time without hemolysis.

Simple modifications and variations of the present invention can be readily used by those skilled in the art, and all such variations or modifications can be considered to be included within the scope of the present invention.

100, 200: device for plasma separation
101, 201: superstructure
102, 202: substructure
103, 203: first opening
104: air outlet
105, 205: second opening
106, 206: Protruding member
111, 211: membrane filter
112, 212: first sheet
113, 213: cavity
114, 214: second sheet
121, 221: frame member
122, 222: border member
123, 223: membrane filter seat
124, 224: first plasma migration pathway
125, 225: second plasma migration pathway
126, 226: plasma chamber
127, 227: communication tube

Claims (26)

An upper structure including at least one first opening formed to inject a blood sample, and a second opening spaced apart from the first opening and connected to the negative pressure applying means at one end side region; And
(i) a membrane filter seat configured to seat a membrane filter for separating plasma from a blood sample injected through the first opening, (ii) a plasma chamber for collecting plasma separated from and in communication with the second opening; (iii) an undercarriage comprising a first plasma transfer passage defined by the bottom surface, the plasma separated by the membrane filter extending to move in the direction of the second opening;
Including,
Here, a blood chamber is formed to receive the blood sample injected between the upper structure and the lower structure by assembling the upper structure and the lower structure,
The membrane filter seating portion defines a width of the first plasma movement passage in a central region of a bottom surface formed by a pair of surfaces inclined symmetrically or asymmetrically downward from both sides of the substructure,
The first plasma migration passage is in communication with the plasma chamber by connecting means,
On the pair of symmetrically or asymmetrically inclined surfaces of the membrane filter seating portion, the first guide protrusion pattern and the second guide protrusion protrude so that the plasma separated through the membrane filter moves while converging to the first plasma movement path by capillary action. Each of the patterns is provided, and
The connecting means includes a second plasma movement passage formed across the thickness of the bottom at or near a downstream end end region of the first plasma movement passage, and a cavity of a shape extending in the longitudinal direction is formed on the bottom surface of the bottom of the lower structure. A plasma transfer channel formed by a combination of a first sheet and a second sheet attached to a lower surface of the first sheet, the plasma transfer channel separating plasma from blood, connecting the second plasma transfer passage and the plasma chamber Device for An upper structure including a first opening formed in one end side region for injecting a blood sample, and a second opening positioned in an end side region in a direction opposite to the first opening and connected to the sound pressure applying means; And
(i) a membrane filter seat configured to seat a membrane filter for separating plasma from a blood sample injected through the first opening, (ii) a plasma chamber for collecting plasma separated from and in communication with the second opening, and (iii) a lower structure comprising a first plasma migration passage inclined downwardly and bounded by a bottom surface such that plasma separated by the membrane filter moves in the direction of the second opening;
Including,
Here, the blood chamber is formed to accommodate the blood sample injected between the upper structure and the lower structure by the assembly of the upper structure and the lower structure,
The membrane filter seating portion defines a width of the first plasma movement passage in a central region of a bottom surface formed by a pair of surfaces inclined symmetrically or asymmetrically downward from both sides of the substructure,
The first plasma migration passage is in communication with the plasma chamber by connecting means,
On the pair of symmetrically or asymmetrically inclined surfaces of the membrane filter seating portion, the first guide protrusion pattern and the second guide protrusion protrude so that the plasma separated through the membrane filter moves while converging to the first plasma movement path by capillary action. Each of the patterns is provided, and
The connecting means includes a second plasma movement passage formed across the thickness of the bottom at or near a downstream end end region of the first plasma movement passage, and a cavity of a shape extending in the longitudinal direction is formed on the bottom surface of the bottom of the lower structure. A plasma transfer channel formed by a combination of a first sheet and a second sheet attached to a lower surface of the first sheet, the plasma transfer channel separating plasma from blood, connecting the second plasma transfer passage and the plasma chamber Device for A pair of first openings having a shape extending in the longitudinal direction to inject a blood sample, and a second opening formed in an end side region in a direction opposite to the first openings and connected to the sound pressure applying means; Superstructure; And
(i) a membrane filter seat configured to seat a membrane filter for separating plasma from a blood sample injected at least once through the first opening; and (ii) a plasma collecting plasma separated from and communicated with the second opening. A lower structure including a chamber and (iii) a first plasma migration passage extending to horizontally move plasma separated by the membrane filter in the direction of the second opening and bounded by a bottom surface;
Including,
Here, a blood space is formed to accommodate the blood sample injected between the upper structure and the lower structure by the assembly of the upper structure and the lower structure,
The membrane filter seating portion defines a width of a first plasma movement passage in a central region of a bottom surface formed by a pair of surfaces inclined symmetrically or asymmetrically downward from both sides of the substructure,
The first plasma migration passage is in communication with the plasma chamber by connecting means,
On the pair of symmetrically or asymmetrically inclined surfaces of the membrane filter seating portion, the first guide protrusion pattern and the second guide protrusion protrude so that the plasma separated through the membrane filter moves while converging to the first plasma movement path by capillary action. Each of the patterns is provided, and
The connecting means includes a second plasma movement passage formed across the thickness of the bottom at or near a downstream end end region of the first plasma movement passage, and a cavity of a shape extending in the longitudinal direction is formed on the bottom surface of the bottom of the lower structure. A plasma transfer channel formed by a combination of a first sheet and a second sheet attached to a lower surface of the first sheet, the plasma transfer channel separating plasma from blood, connecting the second plasma transfer passage and the plasma chamber Device for 4. A device according to any one of the preceding claims, wherein a membrane filter is seated on the membrane filter seat in the substructure. The method of claim 4, wherein the upper structure has a support for fixing the membrane filter protruding while being spaced apart at regular intervals on the back, to ensure that the membrane filter is in close contact with the membrane filter seat when assembled with the lower structure. And a device configured to enhance the capillary phenomenon in the two guide protrusion pattern. The device of claim 2, wherein the upper structure further comprises an air outlet. The device of claim 6, wherein the first opening is a circular cavity, and the cavity is formed in a fused shape of the cavity. The device of claim 7, wherein the first opening has a shape in which air outlets are fused in a wing shape on both sides of the circular cavity. The device according to claim 1, wherein each of the upper structure and the lower structure is integrally formed by a molding process. delete delete 4. A device according to any one of the preceding claims, wherein said connecting means further comprises a communication tube formed in a downward direction of the plasma chamber, said communication tube communicating with said plasma moving channel. The device according to any one of claims 1 to 3, wherein at least one of the membrane filter seat, the first plasma passage, and the connecting means is hydrophilized with at least one of an organic material and an inorganic material. . The compound of claim 13, wherein the organic material is selected from the group consisting of amine, hydroxy, carbonyl or epoxy functional groups, and monomers, dimers and polymers having the functional groups, and
And said inorganic material is at least one metal selected from the group consisting of gold, silver, silicon, aluminum, nickel, iron, copper, manganese, silicon, titanium and chromium or oxides thereof. The device according to any one of claims 1 to 3, wherein at least one of the membrane filter seat, the first plasma passage, and the connecting means is surface treated with Bovine serum albumin (BSA). . The device according to claim 1, wherein the first sheet and the second sheet are each in the form of a film. 17. The device of claim 16, wherein the thicknesses of each of the first sheet and the second sheet are different or equal to each other and range from 20 μm to 1 mm. The device according to claim 1, wherein the upper structure and the lower structure are assembled by a double-sided adhesive tape. The device of claim 1, wherein each of the first guide protrusion pattern and the second guide protrusion pattern is a symmetrical or asymmetrical pattern with each other, and a fixed or atypical pattern. The method of claim 19, wherein each of the first guide protrusion pattern and the second guide protrusion pattern is at least one selected from the group consisting of a comb pattern, a vertical line pattern, a horizontal line pattern, a wave pattern, a dot pattern, a pleated pattern, and a sash pattern. A device characterized in that it is a projecting pattern. 21. The method of claim 20, wherein each of the first guide protrusion pattern and the second guide protrusion pattern is a comb-patterned pattern, and the height of the protrusions constituting the same and the distance between the protrusions are in the range of 0.05 to 0.5 mm and 0.5 to 1 mm, respectively. Device characterized in that. The device according to any one of claims 1 to 3, wherein the sound pressure applied by the sound pressure applying means is in the range of 0.1 to 1 kg / cm 2. The membrane filter of claim 4, wherein the membrane filter is an asymmetric membrane comprising at least one porous medium, the upper region of the asymmetric membrane has a pore size of 5 to 100 μm, and the lower region of the asymmetric membrane is 0.1 to 5 μm. And a pore size.  The device of claim 4, wherein the membrane filter has a thickness in the range of 100 to 1,000 μm. a) mounting a membrane filter on the device according to any one of claims 1 to 3;
b) taking a blood sample and injecting a blood sample at least once through the first opening of the device and passing it through the membrane filter to separate plasma from the blood sample; And
c) after a predetermined time has elapsed, collecting the separated plasma in a plasma chamber by applying a negative pressure by means of a negative pressure applying means connected to the second opening;
Separating the plasma from the blood sample comprising a. The method of claim 25, wherein the negative pressure applying means is a syringe pump or a peristaltic pump.

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