KR101968319B1 - Sample Separation Device Based on Origami Using Filtering Layer - Google Patents

Abstract

In the present embodiments, the target substance is concentrated on a specific region by applying an electric field to the selective ion-permeable layer based on origami, and the target material is concentrated to a desired position through the filtering layer having the micropores A sample separation device capable of separating a target material is provided.

Description

[0001] The present invention relates to a paper folding-based sample separation device using a filtration layer,

The technical field to which this embodiment belongs is a biological sample separation apparatus capable of concentrating a target substance in a specific section and capable of low-cost production.

The contents described in this section merely provide background information on the present embodiment and do not constitute the prior art.

Modern medicine aims not only to extend life span, but also to prolong the healthy lifetime. Therefore, future medicine is not centered on therapeutic medicine, but paradigm is changing by implementing 3P of Preventive Medicine, Predictive Medicine, and Personalized Medicine. Early identification and early treatment of diseases have become important means to achieve this specifically, and biomarkers have been actively studied as a means of achieving this.

A biomarker is a marker that can distinguish between normal or pathological conditions, or can be predicted and objectively measured. Biomarkers include nucleic acid DNA, RNA (genes), proteins, lipids, metabolites, and their pattern changes. In other words, biomarkers such as BCR-ABL gene fusion of chronic myelogenous leukemia, which is a therapeutic target of Gleevec from the simple substance such as blood glucose for diagnosis of diabetes, are biomarkers practically used in clinical practice.

DNA (Deoxyribonucleic Acid) is a genetic material that exists in the nucleus, and a gene is a place where chemical information that determines the kind of protein that an organism produces is stored. The information constituting the human body can be grasped by analyzing the DNA, and various DNA analysis techniques are being researched and utilized for the prevention and treatment of diseases. To analyze disease using DNA, gene amplification technology called PCR (Polymerase Chain Reaction) is used. PCR is a method of repeating heating and cooling with a thermostable DNA polymerase to use a single strand produced by sequential separation of a double strand of DNA as an original to make a new double strand. First, heat the DNA and divide it into two chains . Add a short DNA called "primer" to this and cool it so that the primer binds to the DNA. When an enzyme called DNA polymerase is added to this, the primer part becomes the starting point and the DNA is replicated. The DNA is doubled in one cycle called "heating and cooling". Repeating this dozens of times will cause DNA to multiply in billions of times in about an hour.

Protein is a complex molecule made up of relatively simple molecules called amino acids, and is generally very large in molecular weight. There are about 20 kinds of amino acids that make proteins. These amino acids are connected to each other through chemical bonds to form polypeptides. In this case, the binding of amino acids is referred to as a peptide bond, and the peptide bond is referred to as a polypeptide. Proteins in a broad sense can also be called polypeptides. In general, a polypeptide having a relatively small molecular weight is referred to as a polypeptide, and a protein having a very large molecular weight is referred to as a protein. Such a protein is a representative molecule that constitutes the body of an organism, and is a substance that acts as a catalyst for various chemical reactions in cells and immunity. Proteins are very important organisms that constitute living organisms and participate in in vivo reactions and energy metabolism.

By analyzing the DNA or protein as described above, it is possible to grasp the fuming and progress of the cancer or disease. In particular, for the early diagnosis and treatment of intractable diseases such as cancer, a blood fingerprint analysis technique is known in which an indicator protein showing minute changes in the initial stage of normal cells in the blood is developed into cancer cells. Blood fingerprint analysis is a comprehensive analysis of the metabolism data of metabolites present in the blood of cancer patients, taking into account that metabolites of the human body can change depending on the presence or absence of cancer. . Blood fingerprint analysis can diagnose cancer directly from blood, so it can acquire information quickly.

However, the present technology and devices for protein analysis have difficulties in manufacturing devices by using nanotechnology, and they are relatively expensive and difficult to be popularized. In addition, there is a disadvantage in that a high-sensitivity sensor is required for a protein analyzer or it is difficult to perform accurate analysis with a small amount of samples. In recent years, development of nanotechnology and MEMS (Micro Electro Mechanical Systems) technology has enabled it to rapidly separate and purify necessary materials with only a small amount of sample by patterning it into a single nano structure in a fluid device. Biotechnology and medical engineering are actively engaged in efforts to apply them. In addition, advanced MEMS / NEMS technology enables more precise patterning of nanostructures at desired locations with error tolerances of a few nanometers. This technology is combined with microfluidic channels to provide a micro total analysis system (m- TAS) or lab-on-a-chip.

In particular, a method for realizing biomolecule concentration and separation apparatus capable of enhancing detection accuracy by concentrating biomolecules in a small amount of sample using glass or other heavy and expensive materials is known, And the film is formed so that the target substance can be concentrated at a predetermined position while diffusing through the plate. However, these methods have problems such as difficulty in manufacturing the concentrating and separating device, high cost, large and heavy equipment, and inconvenience in handling.

Korean Patent Publication No. 10-2015-0083722.

It is a main object of the present invention to provide a biosample separating apparatus capable of producing a low cost by concentrating and separating a target substance in a specific region by applying an electric field to a selective ion-permeable layer based on origami.

Embodiments of the present invention have another object of the present invention to compress a target material (object to be collected) to a desired position and to separate a non-target material (separation object) through a filtration layer whose size is controlled by pressing paper .

Other and further objects, which are not to be described, may be further considered within the scope of the following detailed description and easily deduced from the effects thereof.

According to one aspect of the present embodiment, there is provided a coating apparatus comprising: a base including a plurality of base units and configured to fold the plurality of base units; a coating layer disposed in at least a part of the base to prevent adsorption of the sample, A plurality of reservoirs for storing or moving a collection object to be separated from the sample or a separation object not included in the sample, the plurality of reservoirs being located in the plurality of base units with an area set by the coating layer, A selective ion-permeable layer that selectively couples with some of the reservoirs among a plurality of reservoirs to selectively transmit ions, and a selective ion-permeable layer that is positioned in the middle of the movement path of the object to be collected formed by folding the plurality of base units, Provided is a sample separation device including a filtration layer do.

As described above, according to the embodiments of the present invention, it is possible to manufacture the selective ion-permeable layer on the basis of origami, by concentrating and separating the target material in a specific region by applying an electric field to the selective ion-

According to the embodiments of the present invention, there is an effect that the target material can be concentrated and separated at a desired position through the filtering layer in which the size of the micropore is controlled by pressing the paper.

Even if the effects are not expressly mentioned here, the effects described in the following specification which are expected by the technical characteristics of the present invention and their potential effects are handled as described in the specification of the present invention.

1 to 3 are plan views of a biological sample separation apparatus according to embodiments of the present invention.
4 illustrates a slip layer unit in a biological sample separation apparatus according to another embodiment of the present invention.
FIG. 5 illustrates a concentration and separation region of a biological sample separation apparatus according to an embodiment of the present invention.
6A and 6B show a coating layer of a biological sample separation apparatus according to an embodiment of the present invention.
FIG. 7 illustrates a folded state of the biological sample separation device according to an embodiment of the present invention.
8A and 8B illustrate an experimental procedure and a result of a biological sample separation apparatus according to an embodiment of the present invention.
9 shows a folding process of the biological sample separation apparatus according to an embodiment of the present invention.
10A and 10B illustrate a biological sample separation apparatus according to another embodiment of the present invention and simulation results thereof.
11 is a front view of a biological sample separation apparatus including a filtration layer according to another embodiment of the present invention.
12 illustrates the operation of the filtration layer of the biological sample separation device according to another embodiment of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. Will be described in detail with reference to exemplary drawings.

1 shows a biological sample separation apparatus according to an embodiment of the present invention.

In order to facilitate the analysis of a biological sample, the biological sample separation apparatus of this embodiment uses a separation according to the difference in physical properties of components to concentrate the target material and to separate the separation target material. Here, the difference in physical properties is a concept including mobility, mass, charge, size, and the like. In the separation process, ion concentration polarization can be generated using the characteristics of electrophoresis and electro-osmosis, and the sample can be concentrated and separated. The resultant concentrated target substance can be identified by emission spectroscopy such as inductively coupled plasma (ICP). In particular, the present invention can concentrate biomarkers in whole blood of several ng or less several thousand times Therefore, it is easy to judge the reaction even by the naked eye, and early disease detection and diagnosis can be made possible. Examples of target materials include proteins, DNAs, RNAs, steroids, cholesterol, exosomes, and the like, but the present invention is not limited thereto. Depending on the design, samples containing suitable materials may be used.

1, a biological sample separation apparatus 10 according to an embodiment of the present invention includes a base 100, a coating layer 200, a reservoir 300, and a selective ion-permeable membrane 400.

The base 100 includes a plurality of base units which can be folded at predetermined intervals and to be a basic unit to be folded. The base 100 includes a material having at least some fiber texture and the coating layer 200 is additionally bonded to the base 100. The space in which the selective ion- May include at least a part of the non-existent space, and the sample or target material may be moved through the space.

In this embodiment, a paper is used as the base 100. However, the type of the base 100 is not limited to this, and PET (polyethylene terephthalate) is also possible. Particularly, the base 100 is preferably made of a material having a structure capable of bonding with a hydrophobic material or capable of infiltrating a hydrophobic material.

In the case of paper, it is easy to form a coating layer 200 having a uniform thickness on paper in a form of not only being low in cost, good in elasticity and moldability, adsorbing and penetrating with a hydrophobic substance, Do. Such a coating layer has an advantage that it is easy to store a sample and form a channel for moving a certain component.

In the case of paper, it is a hydrophilic material composed of a fiber type. In this case, a capillary is formed according to the fiber structure, and the liquid is moved by the capillary phenomenon when the liquid is dropped. That is, by using such a capillary phenomenon, it is possible to move the liquid even if power is not externally supplied. If the drag force is used, the movement of the separated components can be facilitated for concentration of biomolecules have. In addition to paper, other materials known as fiber-like hydrophilic materials can be used like paper.

Further, according to another embodiment of the present invention, when the concentration is progressed, the concentration plug can be gradually moved when it is concentrated to some extent. In this case, if the drag force can be reversed, it will be very helpful in improving the concentration ratio. This drag force is effective because the capillary phenomenon can be carried out without additional power.

The coating layer 200 is located in at least a part of the base to prevent adsorption of the sample to be treated and to separate the storage space or the movement path.

In the present embodiment, wax or the like may be used as a representative example of the hydrophobic substance as the alcohol fatty acid ester. In this case, wax may be printed on the base 100 or may be bonded by heating. But are not limited to, acrylics, olefins, amides, imides, styrenes, carbonates, vinyl acetals, dienes, vinyl esters, vinyl esters, ketones, fluorocarbons or Teflon ), PDMS, silane, and the like. In addition, there are alkylsilane series silicides and silicone series, such as methylhydrosiloxane-dimethylsiloxane (Methylhydrosiloxane-Dimethysiloxane Copolymer).

Generally, when a material becomes hydrophobic in a material, it may have hydrophobicity due to the influence of the structure of the surface contacting the water and the characteristics of the surface of the material itself. Particularly, in the case of forming the microchannel 20 by coating on the base paper, the latter corresponds to the latter. In forming the channel by coating the wax on the paper, the hydrophilic material, This is to make it hydrophobic. Such a hydrophobic property can achieve a similar effect even when a fiber material similar to paper is used as the base, so that it is also possible to use a fiber-based base in addition to the cellulose paper.

The reservoirs 300 may be exemplified by including 310 to 340 reservoirs as illustrated in the figure. The reservoir is defined by the coating layer and is respectively located in the base unit. The reservoir is contained in the sample to be measured and at least partially adsorbs and stores a target substance to be separated or a substance to be separated excluding the target substance in the sample Provide a travel route. Each of the reservoirs 300 may be embodied in various shapes such as a triangle, a square, and a star, as well as a circular shape as shown in the drawing. The reservoirs 300 included in each base unit The shape is not necessarily the same.

The reservoirs 300 may provide a pathway for storing or moving a moisturized sample, e.g., a protein sample or a conductive liquid (buffer) to be analyzed.

According to an embodiment of the present invention, the reservoirs 300 may be formed as an exposed space on at least one side of the base 100, without being coupled with the coating layer 200.

The reservoir burls 300 described above are physical spaces formed by the coating layer 200 located on the base 100. In this embodiment, a method of bonding a wax, which is a hydrophobic substance, to the base 100, A method for coating a pattern of a substrate is disclosed. There is no particular limitation on the wax patterning method, and it is possible to simply join the paper and the hydrophobic material, or to bond the paper with the hydrophobic material in a permeated state. Preferably, however, It is preferable to bond them to permeate. In the case of paper, it is easy to form a coating layer having a uniform thickness on paper in a form that is low in cost, good in elasticity and moldability, adsorbed and permeated with a hydrophobic substance, and has excellent bonding strength with a hydrophobic coating material.

The base 100 may include a porous membrane. The thickness of the base 100 may be controlled to control the resolution to such an extent that the sample is separated and concentrated. For example, when the base 100 is paper, the resolution can be controlled by differently applying the thickness of the paper to 50 μm, 180 μm, 350 μm, etc., and the size of the pores of the base 100 can be adjusted It is possible to control the efficiency of electrophoresis (EP).

According to an embodiment of the present invention, the sample or buffer to be treated may be pre-wetted to the reservoir 300 and may be folded to allow the separation to proceed. In another embodiment, A sample or a buffer to be subjected to an un-wetted treatment may be dispensed to a slip layer reservoir 630 of a separate slip layer unit 600 to form a folded base Or may be inserted into the injection reservoir 153 of the injection base unit 150, and the base 100 may be folded to be separated.

The selective ion-permeable layer (400) overlaps with at least a part of the reservoir of at least a part of the plurality of reservoirs to selectively transmit ions. The patterned selective ion-permeable film 400 serves as a kind of nanofilter that selectively transmits a proton.

The selective ion-permeable membrane 400 is formed on a Nafion membrane having a predetermined thickness and area according to a pattern directly designated on the adhesive layer 20 by microflow patterning or pipetting membrane.

Here, there is no particular limitation on the method of patterning the selective ion-permeable material. However, as a method for increasing the concentration efficiency of the target material in the microchannel and controlling the concentration ratio, the conventional printing techniques (for example, inkjet printing) To form a selective ion permeable material. However, if it is desired to provide a biological sample separation apparatus 10 that can be easily manufactured at low cost, a pattern can be formed by cutting a previously formed Nafion membrane and attached to the base 100. The selective ion-permeable film 400 may be formed to have a thickness of several hundred nanometers to tens of micrometers, and may be formed in a rectangular pattern having a width of tens to hundreds of micrometers. The selective ion-permeable layer 400 may be formed of a polyelectrolyte such as PSS (Polystyrene Sulfonate) or PAH (Polyallylamine Hydrochloride) in addition to Nafion. .

The selective ion-permeable membrane 400 is a membrane for selectively transmitting a specific ionic material, and is a component for implementing a nanochannel that functions as a nano filter. The selective ion-permeable membrane 400 can function as a nanofilter for selectively transmitting a proton, for example. When the selective ion-permeable membrane 400 is realized as nafion, H + ions are selectively and rapidly permeated by the hopping and vehicle mechanism due to SO3- in the Nafion's chemical structure . Therefore, the selective ion-permeable film 400 can perform the function of a nanofilter.

In one embodiment of the present invention, the sample is easily separated and concentrated. In order to maximize the effect of sample separation due to the potential difference, the reservoir 300 located in the base unit adjacent to the end of the base 100 is provided with a selective ion- 400).

2 is a plan view of a biological sample separation apparatus 10 according to an embodiment of the present invention. Referring to FIG. 1, FIG. 2 will be described as follows.

The first end base unit 110 is a base unit located at one end of the base unit constituting the base 100 and the second end base unit 120 is a base unit located at the other end. The first and second end base units 110 and 120 may be configured to apply a voltage and support the base when the base 100 is folded to separate the sample.

The first end reservoir 310 and the base 100 are separated from each other by the coating layer 200. The first end reservoir 310 and the first end reservoir 310 may be coated with a coating layer 200 containing a hydrophobic substance, The first electrode 510 may be positioned so as to separate and concentrate the sample.

The second terminal base unit 120 may be coated or cut with a coating layer 200 including a hydrophobic material and may be attached to the second terminal reservoir 320 and the base 100 The second electrode 520 may be positioned so as to separate and concentrate the sample.

The third end base unit 130 is a base unit adjacent to the first end base unit 110 and the fourth end base unit 140 is a base unit adjacent to the second end base unit 120.

The third end reservoir 330 is located at the third end base unit 130 and is connected to the first end reservoir 310. The fourth end reservoir 340 is located in the four-end base unit 140 and is connected to the second end reservoir 320. The third and fourth terminal reservoirs 330 and 340 are connected to the selective ion-permeable membrane 400 to allow selective transmission of ions.

The meaning of the connection means that a coating layer 200 is applied or cut between both reservoirs, so that there is an unattached channel. Between the reservoirs 300 connected to each other, the sample, the buffer, and the like can be mutually moved by the capillary phenomenon.

In an embodiment of the present invention, a positive electrode, a negative electrode, or a ground may be connected to each of the first and second reservoirs 310 and 320 so that a potential difference is generated between the first end reservoir 310 and the second end reservoir 320, A potential difference is generated between both ends of the selective ion permeable membrane 400 located at the fourth terminal reservoir 340 connected to the third terminal reservoir 330 and the second terminal reservoir 320 connected to the first terminal reservoir 310 , The sample can be separated and concentrated by ion concentration polarization (ICP).

3 is a plan view of a biological sample separation apparatus 10 according to an embodiment of the present invention. Referring to FIG. 1 and FIG. 2, FIG. 3 will be described below.

The injection base unit 150 may include an injection base unit coating layer 151, an injection reservoir 153, and an injection channel 155.

The injection base unit 150 is directly connected to the base unit other than the first to fourth base units 110, 120, 130 and 140, and when the base 100 is folded, to be.

The injection base unit coating layer 151 is located in at least a part of the injection base unit to prevent adsorption of the sample to be treated and to distinguish the storage space or the movement path.

The injection base unit 150 may be disposed in such a manner that at least a part of the coating layer 200 for preventing adsorption of the sample or the buffer is applied or cut and attached thereto. An injection channel 155 may be positioned to connect the reservoir 153 and the reservoir of the connected base unit to the injection reservoir 153 and to provide a path for the sample to be treated.

According to an embodiment of the present invention, the injection reservoir 153 and the injection channel 155 may be formed on at least one side of the exposed base 100 without being coupled with the coating layer 200.

The injection base unit 150 has a purpose of injecting a sample to be treated or a buffer for wetting the reservoir. In one embodiment of the present invention, after a blood sample or the like is injected through the injection base unit 150, the connected base unit and the injection base unit 150 are cut to prevent the sample from being dispersed again .

FIG. 4 illustrates a slip layer unit 600 in addition to the biological sample separation apparatus 10 of FIG. 2 according to an embodiment of the present invention. Referring to FIG. 1 and FIG. 2, FIG. 4 will be described as follows.

4 shows a slip layer unit 600, which can be additionally configured to the base 100 that can be folded, in the biological sample separation apparatus 10.

The slip layer unit 600 includes a slip layer base 610 to which a sample or the like can be adsorbed, a slip layer coating layer 620 that is positioned at least in a partial region of the slip layer base 610 to prevent adsorption of a sample or the like, And a slip layer reservoir 630 that defines a region by the coating layer 620 and at least partially adsorbs and stores a sample to be measured or provides a movement path. The support unit 640 may further include a support unit 640 that may serve as a handle when inserting or removing the slip layer unit 600 from the base 100. The support unit 640 may include a base 100 or a slip layer unit 600 ) Or a plastic material.

The slip layer unit 600 can sample the slip layer reservoir 630 and insert the slip layer reservoir 630 in a specific section of the base 100 to separate and concentrate the sample. The slip layer unit 600 can wet the sample, The sample or the like may be moistened through the unit 150, inserted into a specific section of the base 100, separated and concentrated to obtain a concentrated target material or a separation target material in a specific section.

FIG. 5 illustrates a concentration and separation region of a biological sample separation apparatus according to an embodiment of the present invention. Referring to FIG. 2, FIG. 5 will be described as follows.

The base unit, which is a basic unit of the base 100, may be configured to selectively remove at least one target material or a separation target material from the sample, if necessary.

The base 100 may be folded at regular intervals. When the base units are overlapped with each other as a basic unit to be folded and a potential difference is generated across the base 100, the sample or the like may be electroosmosis flow through the overlapped reservoirs , EOF) or electrophoresis (EP) and the mobility of sample components. At this time, a target substance or a substance to be separated can be selectively obtained by separating a specific section for one or more base units as necessary.

For example, albumin and globulin among components of plasma are protein components having the highest volume in plasma, and they are separated from each other as noise of a signal. Therefore, in the case where plasma is used as a sample to be treated as an example of the present invention, when plasma is sampled directly to a reservoir or a reservoir, and a potential is applied to both ends of the base 100, plasma Separation of components occurs, and the base unit in which the separated albumin and the globulin are concentrated as shown in Fig. 5 can be separated.

6A and 6B illustrate a method of forming a coating layer of a biological sample separation apparatus according to an embodiment of the present invention.

The coating layer 200 includes a hydrophobic material for preventing the adsorption of the sample and may be attached to both sides of the base 100 after forming a coating film of the hydrophobic material in a predetermined pattern by cutting in a physical cutting manner, In accordance with the pattern described above.

According to one embodiment of the present invention, the coating layer 200 prevents the sample or the like from being adsorbed to the base 100 by being coupled with the base 100, The buffer or other channel can be used to store or move the sample.

6A, the coating layer 200 may be formed by applying a predetermined pattern to the base 100 as shown in FIG. 6B. Alternatively, the coating layer 200 can be formed can do. However, it is preferable to apply the coating material to the reservoirs 300 in order to prevent the loss of the sample or the like.

FIG. 7 shows a folded state of the biological sample separation apparatus 10 according to an embodiment of the present invention. 7 will be described as follows.

The sample separation apparatus 10 includes a base 100 having a structure in which a plurality of base units are folded, a selectively folded structure being released, a base unit 100 disposed in at least a part of each of the base units to prevent adsorption of a sample to be treated A region defined by a coating layer 200 and a coating layer 200 for discriminating a storage space or a moving path and each being located in the base unit and included in a sample to be measured, A plurality of reservoirs 300 for adsorbing and storing at least a part of the substance to be separated except for the at least one reservoir and providing a movement path and a selective ion permeability Layer 400 as shown in FIG.

The sample separation apparatus 10 includes a base 100, and the base 100 is folded so that the base units having a predetermined interval are overlapped with each other. At this time, the first end base unit 110 and the second end base unit 120 can be a support for the entire base 100 without overlapping with other base units. A coating layer 200 patterned in a uniform shape is coupled to the base 100 and a reservoir 300 capable of storing or moving a sample or the like as a base 100 exposed to the base layer, can do.

In the folded base 100, the reservoirs 300 may be embodied so as to overlap with each other, and the specimen or the like may be moved between the overlapped reservoirs 300 at this time. Among the reservoirs, the reservoir 300 located at the first end base unit 110 is the first end reservoir 510 and the reservoir 300 located at the second end base unit 120 is the second end reservoir End reservoir 320. A first electrode 510 may be connected to the first end reservoir 310 and a second electrode 520 may be connected to the second end reservoir 320. According to one embodiment of the present invention, the voltage is applied to the first electrode and the second electrode is grounded so that the sample is separated using the electroosmosis (EOF) and electrophoresis (EP) phenomenon. Can be implemented.

The base unit adjacent to the first end base unit 110 is a third end base unit 130 and the third end reservoir 310 is located and the base unit adjacent to the second end base unit 120 is a fourth end base unit An end base unit 140 and a fourth end reservoir 340 are located. The selective ion permeable membrane 400 for selectively passing ions may be combined with the third end reservoir 330 and the fourth end reservoir 340.

The sample separator 10 includes a first electrode 510 connected to the first end reservoir 310 and a second electrode 520 connected to the second end reservoir 320. The second electrode 520 is connected to the second end reservoir 320, As shown in FIG.

According to an embodiment of the present invention, a sample separating apparatus 10 includes a reservoir 300 positioned at a first end base 110 positioned at one end of a base 100 and serving as a support for the base 100, A terminal reservoir 310, a second end reservoir 320 as a reservoir located at the other end of the base 100 and located in a second end base 120 serving as a base support for the base, A third end reservoir 330 and a second end base unit 120 which are located in the third end base unit 130 adjacent to the unit 110 and are connected to the first end reservoir 310, And a fourth end reservoir 340 which is a reservoir 300 positioned in the fourth end base unit 140 adjacent to the second end reservoir 320 and connected to the second end reservoir 320.

The sample separation apparatus 10 is directly connected to a base unit other than the first to fourth base units 110, 120, 130 and 140 and is located at least partially outside the folded base 100, And may further include a base unit 150.

A coating layer 200 for preventing adsorption of a sample or a buffer may be attached to the injection base unit 150 in such a manner that at least a part of the coating layer 200 is applied or cut and adhered thereto. Which is a movement path of a sample to be treated which is exposed without being coupled to the injection reservoir 153 and the reservoir of the connected base unit and the injection reservoir 153, (Not shown).

The injection base unit 150 has a purpose of injecting a sample to be treated or a buffer for wetting the reservoir 300. As an embodiment of the present invention, after a blood sample or the like is injected through the injection base unit 150, the connected base unit and the injection base unit 150 may be cut out so that the sample is diffused again Can be prevented.

In an embodiment of the present invention, the slip layer unit 600, which is separate from the base 100, may be formed by inserting the slip layer reservoir 630 into a specific section of the folded base 100 A voltage may be applied to the first electrode and the second electrode to separate the sample and the like, and the slip layer unit 600 may be collected to use a sample concentrated or separated in a specific section.

As a matter of the present invention, when voltage is applied to the sample separation apparatus 10, one embodiment of concentration through the selective ion permeable membrane is as follows.

For example, when the blood sample is administered to the reservoirs 300, blood fluid contacts the surface of the selective ion-permeable membrane 400 in the region where the reservoir 300 and the selective ion-permeable membrane 400 are overlapped with each other There are induced charges of different nature between the two. The specific layer in which induced charges generated in the fluid exist is called an electric double layer (EDL). At this time, when external power is applied to the first end reservoir 310 and the second end reservoir 320 disposed at both ends to generate a voltage difference, the ions in the fluid are electrically It is attracted to the electrode opposite to the property. In this way, the ions move between the reservoirs 300 according to their electrical properties, and lead the fluid particles together by the viscous force. Therefore, the entire fluid flow occurs, and the movement of the fluid is called electro-osmosis flow (EOF), and the movement of the ions is called electrophoresis (EP).

Such characteristics of electrophoresis and electro-osmosis vary in characteristics near the nanochannel realized by the selective ion-permeable membrane (ipsm), resulting in ion concentration polarization. Therefore, ion enrichment occurs on the cathode side and ion depletion occurs on the anode side in the reaction region between the selective ion-permeable film 400 functioning as a nano channel and the reservoir 300 superimposed on the membrane. do. At this time, the depletion zone acts as an electric barrier to charged proteins due to the deficient low ion concentration and thus the high electric field. As a result, the protein does not pass through the deficiency area and is concentrated before it. That is, the protein, which is the target substance, is concentrated in a very short time in front of the deficiency region in the reservoirs 300.

FIG. 8 shows an experimental procedure and results using the sample separation apparatus according to an embodiment of the present invention. Referring to FIG. 7, FIG. 8 will be described as follows.

8A shows a state in which no voltage is applied. 7 and 8A, a sample to be treated is applied to each reservoir 300 as an orange G dye (1 mg / ml orange G dye) or a phosphate buffer saline (PBS) solution in a reservoir 300 And the selective ion permeable membrane 400 is placed in the third end reservoir 330 and the fourth end reservoir 340.

FIG. 8B shows a state in which a voltage is applied. 7 and 8B, after the base unit 100 is folded to overlap the areas of the reservoirs 300, the first electrode 510 is connected to the first end reservoir 310, The second electrode 520 is connected to the second end reservoir 320 and then the first electrode 510 is grounded and the voltage of 100V is applied to the second electrode 520 for 10 minutes. (100). ≪ / RTI > At this time, ion concentration polarization (ICP) is generated as an effect of the selective ion-permeable film 400, and as a result, the closer to the base unit to which V + is applied, the thicker the concentration of the sample and the depletion area of the sample and the concentration area it can be confirmed that the separation occurs so that the preconcentrated area is clearly distinguished.

9 shows a folding process of the biological sample separation apparatus according to an embodiment of the present invention.

The base 100 may be provided to be foldable at predetermined intervals. In this case, as shown in FIG. 9, the levers 300 located in the respective base units are folded by the pleat folds so that the adjacent levers 300 As shown in Fig. Each of the reservoirs 300 may be embodied in various shapes such as a triangle, a square, and a star, as well as a circular shape as shown in the drawing. The reservoirs 300 included in each base unit The shape is not necessarily the same.

The position where the collected object is concentrated among the plurality of reservoirs may vary depending on the intensity of the electric field, the length of the moving channel, the thickness of the base unit, and the size of the micropore. The Origami-chip or VFAs (Vertical Flow Assays) structure can reduce the reaction time, line interferences, and especially the Hook effect, compared to the conventional LFAs (Lateral Flow Assays) have.

In particular, you can simply add layers in the middle of layers. That is, a plurality of reservoirs may be formed for each layer or one layer to selectively detect different targets.

10A and 10B illustrate a biological sample separation apparatus according to another embodiment of the present invention and simulation results thereof. 1 to 9, the following will be described.

FIG. 10A illustrates a sample separation apparatus according to an embodiment of the present invention. In FIG. 10A, the sample separation apparatus 10 shown in FIGS. 1 to 9 includes a plurality of reservoirs And a plurality of sample separation channels. In the sample separation apparatus 10, the plurality of reservoirs 300 can form a sample separation channel adjacent to each other when the sample separation apparatus 10 is folded at intervals of the base units. At this time, as shown in FIG. 10A, each of the unit base units includes a plurality of reservoirs 300, and a plurality of sample separation channels can be formed, and different samples can be simultaneously separated and concentrated. The sample separation apparatus 10 shown in Fig. 10A can be folded and includes an A sample separation channel, a B sample separation channel, a C sample separation channel, and a D sample separation channel.

FIG. 10B illustrates the results of simulation for different samples using the sample separation apparatus 10 having a plurality of sample separation channels shown in FIG. 10A. As shown in FIG. 10A, sample separation and concentration occur when a V + voltage and a ground are applied to both ends of a sample separation apparatus 10 having a plurality of sample separation channels, Different separation and concentration results are obtained for ~ D. The dotted line indicates the unit interval of the sample separator which can be folded, and corresponds to the interval of the base unit of the present invention.

11 is a front view of a biological sample separation apparatus including a filtration layer according to another embodiment of the present invention. The sample separation apparatus includes a base, a coating layer, a plurality of reservoirs, a selective ion-permeable layer, and a filtration layer.

The base includes a plurality of base units and a plurality of base units 1110 are formed to be folded. At least some of the plurality of base units may be separated.

The coating layer is located in at least a part of the base to prevent adsorption of the sample and to distinguish the storage space or the movement path.

The plurality of reservoirs are located in a plurality of base units with areas set by the coating layer. The plurality of reservoirs store or move the collection object to be separated from the sample. Or the plurality of reservoirs stores or moves the separated object not included in the sample.

The first end base unit 1150 located at one end of the base includes a first end reservoir and the second end base unit 1160 located at the other end of the base includes a second end reservoir. The sample separation apparatus may further include a first electrode connected to the first end base unit and a second electrode connected to the second end base unit.

The selective ion-permeable layer combines with some of the reservoirs among the plurality of reservoirs to selectively transmit ions. The number of the selective ion-permeable layers may be plural. The plurality of selective ion-permeable layers 1130 and 1140 may be separated at predetermined distances according to the direction of movement of the object to be collected. The plurality of selective ion-permeable layers are connected to the first end reservoir and the second end reservoir, respectively.

An electric field (1190) is applied to the plurality of selective ion-permeable layers to concentrate the collected object in the middle of the movement path of the collection object formed by folding the plurality of base units. The sample separation apparatus forms a concentration region 1104 in the sample separation channel, i.e., the movement path of the collection object formed by folding a plurality of base units.

The filtration layers 1101 to 1103 are located in the middle of the movement path 1120 of the collection object formed by folding the plurality of base units 1110 to filter the separation object. The filtration layer comprises a porous membrane, and the size of the micropores in the filtration layer can be predetermined. The size of the pores of the filtration layer is larger than the size of the object to be collected and smaller than the size of the object to be separated.

The sample separation device inserts or folds the filter layer having a predetermined size into the sample separation channel according to the size of the object to be collected, thereby forming a sample separation channel that can be separated stepwise. That is, the sample separation apparatus can separate the object to be collected and the object to be separated.

The filtration layer is directly connected to the base unit that is not located at the distal end among the plurality of base units and the filtration layer is inserted in the middle of the movement path of the collection object in the process of folding the plurality of base units. For example, the injection base unit 150 illustrated in FIG.

On the other hand, the filtration layer may be formed such that it can be inserted into the middle of the overlapped base units such as the slip layer unit illustrated in Fig. The filtration layer may include a handle to facilitate insertion.

The number of the filtration layers is a plurality and the plurality of filtration layers are located on the movement path of the collection object formed by folding the plurality of base units to separate the two or more separation objects step by step.

The filtration layer may further include a display portion for indicating the intensity of the pressure or the size of the micro pores applied to the filtration layer.

The position at which the object to be collected is concentrated may depend on the field strength, the length of the moving channel, the thickness of the base unit, the size of the micropores in the base, the size of the micropores in the filtration layer, and the location of the filtration layer. The sample separation apparatus can separate the object to be separated in advance on the movement path of the object to be collected and adjust the concentration position of the object to be collected.

The method of pressing the membrane (e.g., the base unit or the filtration layer) can be simply performed using a hand press machine. The membrane is fixed on a sensor that measures the strength of the pressure, such as a load cell. You can observe the pressure applied through the pressure indicator connected to the load cell and observe the physical change of the membrane with the pressure intensity.

The pressure indicator is a device that converts analog pressure intensity to digital signal. The pressure indicator indicates that the thickness and pore size of the paper vary depending on the magnitude of the pressure applied to the paper. It can be seen that the size of the fine holes decreases remarkably according to the intensity of the pressure. For example, the micropores of the squeezed paper can be analyzed through a scanning electron microscope (SEM).

12 illustrates the operation of the filtration layer of the biological sample separation device according to another embodiment of the present invention.

Referring to FIG. 12, a plurality of filtration layers and a concentrated region may be formed in the middle of the separation channel of the biological sample separation apparatus. For example, the plurality of filtration layers may form a first filtration layer 1101, a second filtration layer 1102, a third filtration layer 1103, and a concentration region 1104. The plurality of filtration layers 1101, 1102, and 1103 have different micropore sizes using mechanical or physical methods or using different membranes that have been commercialized.

Applying an electric field from the left to the right will move the target material from left to right as a whole. That is, an electric field is applied to the selective ion-permeable layer to concentrate a plurality of objects, and at the same time, a plurality of objects are separated without passing through some filtration layers or passing through some filtration layers.

If the size of the micropores in the first filtration layer 1101 is 10 μm, the material A larger than 10 μm is filtered. If the size of the micropores in the second filtration layer 1102 is 1 μm, the material B larger than 1 μm is filtered If the size of the micropores in the third filtration layer 1103 is 100 nm, the material C larger than 100 nm is filtered, and finally the material D is concentrated to form the humped area 1104. A, B, and C are separated gradually by membranes with different pore sizes, and eventually only the smallest D particles (eg, exosomes) can be isolated . As a result, only pure D-targets can be isolated and concentrated. By filtering the separation objects A, B, and C on the separation channel using the filtration layer, the total number of base units can be reduced, the concentration area can be adjusted, and the concentration time can be reduced.

The base unit including the concentrated collection object may include a perforated line having a small hole for easy separation from the base.

The present embodiments are for explaining the technical idea of the present embodiment, and the scope of the technical idea of the present embodiment is not limited by these embodiments. The scope of protection of the present embodiment should be construed according to the following claims, and all technical ideas within the scope of equivalents thereof should be construed as being included in the scope of the present invention.

10: Sample separation apparatus 100: Base
110, 120, 130, 140: end base unit
150: Injection base 153: Injection reservoir
155: Injection channel 200: Coating layer
300: reservoirs 310, 320, 330, 340:
400: selective ion-permeable layer 510, 520: electrode
600: Slip layer unit 610: Slip layer base
611: slip layer coating layer 630: slip layer reservoir
620: supports 1101, 1102, 1103: filtration layer
1104:

Claims (9)

A base including a plurality of base units that can be folded at predetermined intervals and to be a basic unit to be folded;
A coating layer positioned in at least a part of the base to prevent adsorption of the sample and distinguish the storage space or the movement path;
A plurality of reservoirs located in the plurality of base units with an area set by the coating layer, for storing or moving a collection object to be separated from the sample or a separation object not included in the sample;
A selective ion-permeable layer that selectively couples with some of the reservoirs among the plurality of reservoirs to selectively transmit ions; And
A plurality of base units disposed in the middle of the movement path of the collection object formed by folding the plurality of base units,
. The method according to claim 1,
The base includes:
Wherein at least some of the plurality of base units are separated from each other. The method according to claim 1,
Wherein the number of the selective ion-permeable layers is a plurality, and the plurality of selective ion-permeable layers are separated and positioned according to a moving direction of the object to be collected,
Wherein an electric field is applied to the plurality of selective ion-permeable layers to concentrate the object to be collected in a movement path of the object to be collected formed by folding the base. The method of claim 3,
Wherein the first end base unit located at one end of the base includes a first end reservoir and the second end base unit located at the other end of the base includes a second end reservoir,
Wherein the plurality of selective ion-permeable layers are connected to the first end reservoir and the second end reservoir, respectively. 5. The method of claim 4,
Further comprising a first electrode connected to the first end base unit and a second electrode connected to the second end base unit. The method according to claim 1,
Wherein the filtration layer comprises a porous membrane,
Wherein the size of the pores of the filtration layer is larger than the size of the object to be collected and smaller than the size of the object to be separated. The method according to claim 6,
The size of the micropores is controlled by pressing the filtration layer,
Wherein the filtration layer further comprises a display unit for indicating the intensity of the pressure applied to the filtration layer or the size of the micropore. The method according to claim 1,
Wherein the number of the filtration layers is a plurality and the plurality of filtration layers are located on a movement path of the collection object formed by folding the base so that two or more separation objects are separated stepwise. The method according to claim 1,
Wherein the filtration layer is directly connected to a base unit not located at an end of the plurality of base units,
Wherein the filter layer is inserted in the middle of the movement path of the object to be collected during the folding of the base.

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