KR101718951B1 - Biomolecular preconcentration device and fabrication method thereof - Google Patents

Abstract

The present invention relates to an apparatus for concentrating biomolecules, and a method for manufacturing the same. The apparatus can be manufactured at low costs by using a significantly simple method where substances such as cellulose paper and a selective ion transmission membrane forming a membrane, a reservoir, and a channel are cut to be attached.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a biomolecule concentration apparatus,

The present invention relates to a biomolecule thickening apparatus and a method of manufacturing the same, and more particularly, to a biomolecule thickening apparatus and a method of manufacturing the same, in which a biomolecule thickening apparatus capable of producing a low- And a manufacturing method thereof.

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 are becoming very important means to achieve this specifically, and biomarkers have been actively studied as a means to achieve 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 a cell and immunity (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 (μ- TAS) or lab-on-a-chip.

In particular, a method of realizing a biomolecule concentration apparatus capable of enhancing detection accuracy by concentrating biomolecules in a small amount of sample by using glass or other heavy and expensive materials is known. This is because the analyte is a narrow tube or a thin plate And the film is formed so as to concentrate the target material at a predetermined position while diffusing the target material. However, these methods are problematic in that it is difficult to manufacture a thickening apparatus, a large amount of money is put into the apparatus, a thickening apparatus is large and heavy, and handling is inconvenient.

Korean Patent Publication No. 10-2015-0083722 (published on Aug. 20, 2015)

It is an object of the present invention to provide a very simple and economical method of cutting and attaching materials such as a membrane, a reservoir, a channel, etc., an ion perm-selective membrane, a cellulose paper, Which can be produced at low cost.

Another object of the present invention is to provide a method for manufacturing a biomolecule concentration apparatus for achieving the above object.

According to an aspect of the present invention, there is provided an apparatus for concentrating biomolecules comprising: two patterned selective ion-permeable membranes formed on a straight line; And a porous membrane formed between the selective ion-permeable membranes and patterned at both ends, wherein the porous membrane formed at both ends of the selective ion-permeable membranes is a buffer reservoir, the porous membrane formed between the selective ion- The membrane is characterized by being a reaction membrane.

And the buffer reservoir and the reaction membrane are formed such that their ends partially overlap each other on the side in contact with the selective ion-permeable membranes.

The buffer reservoir and the reaction membrane are formed by patterning by cutting or by patterning using a hydrophobic material.

The selective ion-permeable membranes may be attached after forming a pattern by cutting, or formed by a technique of microflow patterning or pipetting.

Power is applied to each of the buffer reservoirs at both ends to concentrate biomolecules on the reaction membrane, and a sample sample containing the biomolecule is administered to the reaction membrane.

A channel formed from the porous membrane and connected to the portion extending from the reaction membrane; And a sample injection membrane formed of the porous membrane on the other side of the channel and formed in a predetermined shape having a portion having a larger width than the width of the channel.

And power is applied to each of the buffer reservoirs at both ends to concentrate the biomolecules on the reaction membrane, and a sample sample containing the biomolecule is administered to the sample injection membrane.

A portion extending from the reaction membrane on both side ends of the channel, and a portion of the sample injection membrane.

The reaction membrane, the channel, and the sample injection membrane are integrally formed.

The biomolecule concentration apparatus further comprises an ion-permeable membrane on the pre-designated substrate and an adhesive layer for fixing the porous membrane.

The buffer reservoir and the reaction membrane are made of cellulose paper.

According to another aspect of the present invention, there is provided a method of manufacturing a biomolecule concentration apparatus, comprising: forming two patterned selective ion-permeable membranes on a straight line to be spaced apart from each other; And forming a patterned porous membrane between the selective iontophoretic membranes and one at each end, wherein the porous membrane formed at both ends of the selective iontophoretic membranes is a buffer reservoir, and the selective iontophoretic membrane The porous membrane formed is a reaction membrane.

Therefore, the biomolecule concentration apparatus of the present invention can be used as a biomolecule concentrating apparatus without any need of patterning or wetting a hydrophobic substance such as wax-patterning, dipping, soaking, It is a very simple way to cut and paste porous membranes such as ion perm-selective membrane and cellulose paper that make reservoirs and channels, It is possible to manufacture.

In the biomolecule concentration apparatus of the present invention, the reaction membrane may be formed by immobilizing proteins or antibodies in various ways (e.g., inkjet printing, pipetting, soaking, etc.) and separating plasma from whole blood. It is possible to provide a highly sensitive concentrator capable of real-time field diagnosis by concentrating the protein as an antigen in the sample. In the conventional paper-based concentrator, there is a limitation in discriminating the positive or negative reaction of the reaction membrane region of the colorimetric assay visually. However, the present invention is not limited to a method of detecting a protein (biomarker) It is easy to identify the reaction even with the naked eye, and early disease detection and diagnosis are possible.

Further, in the biomolecule concentration apparatus of the present invention, it is possible to expand biomolecules in a high concentration in a short period of time, so that it can be easily expanded by a multiplexing analysis technique, and a living body such as a protein in human serum or whole blood It can be used for concentration of molecules and can be easily implemented in the form of a cartridge such as a rapid kit such as a pregnancy diagnostic kit or a blood glucose sensor through an appropriate housing and interlocked with a mobile communication terminal such as a smart phone Technology can be conveniently applied.

1 is a side view of a biomolecule concentration apparatus according to an embodiment of the present invention.
Fig. 2 shows an exploded perspective view and a top view for explaining the biomolecule concentration apparatus of Fig. 1. Fig.
FIG. 3 is a view for explaining a manufacturing method of the biomolecule concentration apparatus of FIG. 1;
Fig. 4 shows an embodiment of the biomolecule concentration apparatus of Fig. 1 and an experimental example of actual concentration operation.
5 shows a biomolecule concentration apparatus according to another embodiment of the present invention.
6A and 6B show results of experiments on serum samples of different conditions using the biomolecule concentration apparatus of the present invention.

In order to fully understand the present invention, operational advantages of the present invention, and objects achieved by the practice of the present invention, reference should be made to the accompanying drawings and the accompanying drawings which illustrate preferred embodiments of the present invention.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the preferred embodiments of the present invention with reference to the accompanying drawings. However, the present invention can be implemented in various different forms, and is not limited to the embodiments described. In order to clearly describe the present invention, parts that are not related to the description are omitted, and the same reference numerals in the drawings denote the same members. Throughout the specification, when an element is referred to as "including" an element, it does not exclude other elements unless specifically stated to the contrary.

FIG. 1 is a side view of a biomolecule concentration apparatus according to an embodiment of the present invention, and FIG. 2 is an exploded perspective view and a top view for explaining a biomolecule concentration apparatus of FIG.

1 and 2, a biomolecule concentration apparatus 100 according to the present invention includes a substrate 10, an adhesive layer 20 such as a double-faced tape adhered on the substrate 10, / RTI > are formed and patterned one at each end and between two patterned selective ion-permeable membranes (30) and two patterned selective ion-permeable membranes (30) formed / Wherein the patterned porous membranes 40 have respective ends that are in contact with the selective iontransmitting membranes 30 partially overlapping the selective iontophoretic membranes 30, overlap. The attached paper at both ends of the porous membranes 40 are buffer reservoirs 41 and 42 and the paper formed / attached at the center among the porous membranes 40 is a reaction membrane 45 .

Here, the porous membrane 40 may be formed of cellulose paper as a representative example, or a porous membrane may be formed of other materials capable of forming a passive capillary force. In the following, the case where the selective iontophoretic membrane 30 is two patterns is described as an example, but it is noted in advance that a larger number may be attached in a pattern on a line in some cases.

The substrate 10 is not a component for performing the operation for concentration of biomolecules but serves as a supporting plate for allowing the biomolecule concentration apparatus 100 to maintain a stable shape. Thus, a variety of materials that can serve as support plates can be used as the substrate 10. The substrate 10 may be a glass substrate, a plastic substrate such as polycarbonate (PC) or polydimethylsiloxane (PDMS), a silicon substrate, or the like.

The adhesive layer 20 is not a component for performing biomolecule enrichment but attaches the selective ion-permeable membrane 30 and the porous membrane 40, which perform an actual biomolecule concentration function, to the substrate to maintain a stable shape without shaking do. Various materials capable of attaching and fixing the selective ion-permeable membrane 30 and the porous membrane 40 to the substrate 10 may be used as the adhesive layer 20. However, in consideration of convenience in use and cost, It is assumed that a double-sided tape is used as the adhesive layer 20.

The selective ion permeable membrane 30 may be formed by a Nafion membrane having a predetermined thickness and length according to a predetermined pattern directly applied to the adhesive layer 20 using microflow patterning or pipetting techniques . However, the present invention is characterized by providing a biomolecule concentration apparatus that can be easily manufactured at low cost, and therefore it is preferable to cut a previously formed Nafion membrane to form a pattern and adhere to the adhesive layer 20 . The selective iontophoretic membrane 30 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 membrane 30 may be formed of polyelectrolyte such as polystyrene sulfonate (PSS) or polyallylamine hydrochloride (PAH) in addition to Nafion, and may be formed of any material that is selectively permeable to ions .

The selective ion-permeable membrane 30 is a membrane for selectively transmitting a specific ion material, and is a component for implementing a nano channel that performs a function of a nano filter. The selective ion permeable membrane (ipsm) can function as a nanofilter for selectively transmitting a proton, for example. When the selective ion-permeable membrane 30 is implemented as a nafion, H + ions in the Nafion's chemical structure, due to SO3-, may cause hopping and vehicle mechanism (K.D. Kreuer, Chem. Mater. 1996, 8, 610-641). Accordingly, the selective ion-permeable membrane 30 can perform the function of a nanofilter.

The buffer reservoirs 41 and 42 attached to both ends of the porous membranes 40 receive an external voltage through the electrodes to generate a potential difference between the selective ion permeable membranes 30 and the porous membranes 40. As shown in FIG. 2 (b), the buffer reservoirs 41 and 42 are connected to a first voltage (V1, for example, a ground voltage) and a second voltage V2 may be applied, but a thin film electrode may be provided in advance to be connected to an external power source.

On the other hand, the reaction membrane 45 attached between the selective ion-permeable membranes 30 is partially overlapped and attached so as to overlap with the selective ion-permeable membrane 30 at both ends. A sample sample such as whole blood or serum can be administered to the reaction membrane 45 and the substances other than the target substance among the various substances included in the sample sample can be released to the buffer reservoir 41 or 42, Protein) is concentrated on the reaction membrane 45.

For example, when the blood sample is administered to the reaction membrane 45, in the region where the reaction membrane 45 and the selective ion permeable membrane 30 overlap, the blood fluid contacts the surface of the selective ion permeable membrane 30 Induced charges of different nature occur between the two. A specific layer in which induced charges generated in the fluid exist is called an electric double layer (EDL). At this time, when an external power source is applied to the buffer reservoirs 41 and 42 disposed at both ends to generate a voltage difference, the ions in the fluid are attracted to the electrode opposite to the electrical properties of the ions due to the electric field corresponding to the voltage difference . In this way, the ions move in the reaction membrane 45 according to their electrical properties, and lead the fluid particles together by the viscous force. Therefore, the flow of the whole fluid occurs, and the movement of the fluid is called electro-osmosis flow (EOF), and the movement of ions is called electrophoresis (EP).

The characteristics of capillary electrophoresis and electro-osmosis vary depending on the nanochannel implemented by the selective ion-permeable membrane (ipsm), resulting in ion concentration polarization (ICP) . Therefore, ion depletion occurs on the cathode side and ion enrichment occurs on the anode side in the reaction region of the selective ion-permeable membrane 30 functioning as a nano channel and the reaction membrane 45 disposed in an overlapped manner 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 target protein, is concentrated in a very short time before the deficiency region in the reaction membrane 45. Since the size of the depletion region due to the ion concentration polarization expands as the ion concentration polarization of the sample progresses, the protein as the target substance is concentrated in the reaction membrane 45 at a region other than the ends of the iontophoresis membrane 30, do.

FIG. 3 is a view for explaining the production method of the biomolecule concentration apparatus 100 of FIG.

3, an appropriate substrate 10 made of glass, plastic, silicon, or the like is prepared and an adhesive layer 20 such as a double-sided tape is formed on the substrate 10, Fixed.

Next, as in step S200, two patterned selective ion-permeable membranes 30 are formed or attached so as to be spaced apart from each other on a straight line on the adhesive layer 20. [ The selective ion permeable membrane 30 is made of a material such as polyelectrolyte such as Nafion, polystyrene sulfonate (PSS), and polyallylamine hydrochloride (PAH). In addition, any material capable of selectively ion- . It is preferable that the selective ion-permeable membrane 30 is formed by cutting a membrane of the material having a predetermined thickness, width, and length, and is attached to the adhesive layer 20. In addition, the selective ion-permeable membrane 30 may be formed to have a predetermined pattern using a technique such as microflow patterning or pipetting.

Next, as in step S300, porous membranes 40 are formed or attached between the two patterned selective iontransmitting membranes 30 and patterned on both ends thereof. At this time, the patterned porous membranes 40 are formed or attached so that their ends partially overlap the selective ion-permeable membrane 30 on the side in contact with the selective ion-permeable membranes 30. The attached paper at both ends of the porous membranes 40 acts as buffer reservoirs 41 and 42 and the paper attached to the center of the porous membranes 40 is fed to a reaction membrane 45, . It is preferable that the cellulose paper corresponding to the porous membrane 40 is cut to have a predetermined thickness, width, and length to form and attach the pattern. Alternatively, the porous membranes 40 may be formed by a patterning technique using a hydrophobic material (e.g., wax, teflon, toluene, silane, etc.).

Thereafter, as in step S400, a first voltage (e.g., a ground voltage) and a second voltage V2 having different voltage levels are applied to the buffer reservoirs 41 and 42, A sample sample such as whole blood or serum can be administered to concentrate biomolecules.

FIG. 4 shows an embodiment of the biomolecule concentration apparatus 100 of FIG. 1 and an experimental example of actual concentration operation. 4 is a photograph of a state in which a sample sample is injected into the reaction membrane 45, 420 in FIG. 4 is a sample in which a sample sample is injected and a first voltage V2 and a second voltage V2 are applied to the buffer reservoir 41, (E.g., protein) in the reaction membrane 45 after 3 minutes of application.

The first voltage V1 and the second voltage V2 having different voltage levels are applied to the buffer reservoirs 41 and 42 as shown in FIG. Serum, and the like, according to principles such as electrophoresis (electrophoresis) and electro-osmosis as described above, substances other than the target substance among the various substances included in the sample sample are stored in the buffer The target substance (e.g., protein) can be concentrated on the reaction membrane 45 while escaping into the reservoirs 41 and 42. At this time, as described above, the target substance is concentrated in the middle region, not at one side of the reaction membrane 45, as shown at 420, since it is concentrated before the deficiency region.

In the present invention, a biomarker, which is a target substance, can be separated through ion concentration polarization (ICP) and concentrated more than a thousand times in whole blood of several ng or less as a sample, Early disease detection and diagnosis can be confirmed.

FIG. 5 shows a biomolecule concentration apparatus 200 according to another embodiment of the present invention.

Referring to FIG. 5, the biomolecule concentration apparatus 200 according to another embodiment of the present invention includes a substrate 10, an adhesive layer 20, selective ion-permeable membranes 30, porous membranes (40). The porous membranes 40 further include a channel 48 and a sample injection membrane 49 in addition to the buffer reservoirs 41 and 42 and the reaction membrane 45.

That is, the reaction membrane 45 has a portion (or protrusion portion) partially extending in a direction perpendicular to the direction overlapping the selective iontophoretic membrane 30, A part of which is overlapped with or overlapped with one end of one end portion of the frame. The sample injection membrane 49 is formed or attached such that a portion (e.g., a projection) over the other end of the channel 48 overlaps. The sample injection membrane 49 may be formed in a predetermined shape (e.g., a rectangular or circular shape) having a portion having a larger width than the width of the channel 48, As shown in FIG. 5, may include a protruding portion for a portion overlapping with the channel 48.

The porous membranes 40 including the buffer reservoirs 41 and 42, the reaction membrane 45, the channel 48 and the sample injection membrane 49 may all be of the same predetermined thickness, for example, (cellulose paper).

In order to manufacture the biomolecule concentration device 200, the buffer reservoirs 41 and 42 and the channel 48 are formed / attached in advance in the process of forming / attaching the porous membranes 40, (45), and a sample injection membrane (49) at the corresponding positions as described above. Here, the buffer reservoirs 41 and 42 may be formed / attached after forming / attaching the channel 48, the reaction membrane 45, and the sample injection membrane 49.

However, the reaction membrane 45, the channel 48, and the sample injection membrane 49 may be integrally formed / attached without previously forming / attaching the channel 48 in some cases. At this time, it is also possible to form / attach the buffer reservoirs 41 and 42 and then form / attach the integrated type, or to form / attach the buffer reservoirs 41 and 42 after forming / have.

The reaction membrane 45, the channel 48 and the sample injection membrane 49 formed of a porous membrane 40 such as a cellulose paper may be attached after patterning by cutting as described above , Or may be formed by patterning using a hydrophobic substance.

5, the first voltage (V1, for example, ground) and the second voltage (V2) having different voltage levels are applied to the buffer reservoirs 41 and 42, Upon administration of the sample sample, the sample sample flows along the channel 48 to the reaction membrane 45. In the reaction membrane 45, substances other than the target substance among the various substances included in the sample sample are separated by a buffer reservoir (not shown) according to the principle of electrophoresis (electrophoresis) and electro-osmosis as described above 41, 42). So that the target substance (e.g., protein) can be concentrated on the reaction membrane 45. In the present invention, a biomarker, which is a target substance, can be separated through ion concentration polarization (ICP) and concentrated more than a thousand times in whole blood of several ng or less as a sample, Early disease detection and diagnosis are possible.

FIG. 6 shows the results of conducting experiments on serum samples of different conditions using the biomolecule concentration apparatus (100/200) of the present invention.

6 (a) is a result of comparison of the concentration of the protein concentrated in the reaction membrane 45 after 10 minutes with the addition of 2 μl of 0.4 × human serum sample, and the concentration of the protein by emission spectrometry. (a) shows the relationship between the maximum light emission intensity (peak) when applying a voltage of 100 V and the average light emission intensity (average) when a voltage of 50 V to 100 V is applied between the buffer reservoirs 41 and 42 in a predetermined step The luminescence intensity (mean) and the concentration concentration are shown. it can be seen that the concentration (15.5 μM) after 10 minutes at the maximum luminescence intensity peak is 10 times higher than the concentration (1.55 μM) at the start of the sample administration, as shown in FIG.

(b) is a result of comparison of the concentration of the protein concentrated in the reaction membrane 45 after 10 minutes with the concentration of the luminescence intensity and the concentration of the protein by emission spectroscopic analysis, after 2 μl of 1.0 × human serum sample was administered. a voltage of 50 V is applied between the buffer reservoirs 41 and 42 after 10 minutes by the concentration of 4.65 mu M when a voltage of 75 V is applied between the buffer reservoirs 41 and 42 as shown in Fig. (1.55 μM) at the time of the experiment.

As described above, the biomolecule concentration apparatus (100/200) of the present invention can be applied to a conventional method of patterning or wetting a hydrophobic substance such as wax-patterning, dipping, soaking or the like A porous membrane such as an ion perm-selective membrane and a cellulose paper, which form a membrane, a reservoir, a channel, and the like, Cutting and attaching is very simple and low cost manufacturing is possible. In addition, in the biomolecule concentration apparatus (100/200) of the present invention, the reaction membrane 45 is formed by immobilizing proteins or antibodies in various ways (e.g., inkjet printing, pipetting, soaking, It is possible to provide a highly sensitive concentrator capable of real-time field diagnosis by concentrating proteins that become antigens in the plasma. In the conventional paper-based concentrator, there is a limitation in discriminating the positive or negative reaction of the reaction membrane region of the colorimetric assay visually. However, the present invention is not limited to a method of detecting a protein (biomarker) It is easy to identify the reaction even with the naked eye, and early disease detection and diagnosis are possible.

In the biomolecule concentration apparatus (100/200) of the present invention, biomolecules can be highly concentrated in a short period of time. Therefore, it is easy to expand by a multiplexing analysis technique, and human serum or whole blood, And can be used for enrichment of biomolecules such as proteins in a blood vessel or the like and can be easily implemented in the form of a cartridge such as a rapid kit such as a pregnancy diagnostic kit or a blood glucose sensor through an appropriate housing, It is possible to conveniently combine the interworking technique with the communication terminal.

As described above, the present invention has been described with reference to particular embodiments, such as specific elements, and specific embodiments and drawings. However, it should be understood that the present invention is not limited to the above- Those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the essential characteristics of the invention. Accordingly, the spirit of the present invention should not be construed as being limited to the embodiments described, and all technical ideas which are equivalent to or equivalent to the claims, as well as the following claims, are deemed to be included in the scope of the present invention .

Substrate (10)
The adhesive layer (20)
The selective ion permeable membrane (30)
The porous membrane (40)
The buffer reservoirs 41 and 42,
The reaction membrane (45)
Channel 48
Sample Injection Membrane (49)

Claims (22)

Two patterned selective iontophoretic membranes formed in a line and spaced apart from each other; And
And a porous membrane formed between the selective ion-permeable membranes and patterned at both ends,
Wherein the porous membrane formed at both ends of the selective ion-permeable membranes is a buffer reservoir, and the porous membrane formed between the selective ion-permeable membranes is a reaction membrane. The method according to claim 1,
Wherein the buffer reservoir and the reaction membrane are formed such that the ends of the buffer reservoir and the reaction membrane partially overlap each other at a side in contact with the selective ion-permeable membranes. The method according to claim 1,
Wherein the buffer reservoir and the reaction membrane are formed after pattern formation by cutting or by patterning using a hydrophobic substance. The method according to claim 1,
Wherein the selective ion-permeable membranes are attached after patterning by cutting, or formed by a technique of microflow patterning or pipetting. The method according to claim 1,
Wherein power is applied to each of the buffer reservoirs at both ends to concentrate biomolecules on the reaction membrane, and a sample sample containing the biomolecule is administered to the reaction membrane. The method according to claim 1,
A channel formed from the porous membrane and connected to the portion extending from the reaction membrane; And
A sample injection membrane formed of the porous membrane on the other side of the channel and formed in a predetermined shape having a portion having a width larger than the width of the channel,
Wherein the biomolecule concentrating device further comprises: The method according to claim 6,
Wherein power is applied to each of the buffer reservoirs at both ends to concentrate biomolecules on the reaction membrane, and a sample containing the biomolecule is administered to the sample injection membrane. The method according to claim 6,
And a portion extending from the reaction membrane on both side ends of the channel and a portion of the sample injection membrane. The method according to claim 6,
Wherein the reaction membrane, the channel, and the sample injection membrane are integrally formed. The method according to claim 1,
Wherein the biomolecule concentration apparatus further comprises an adhesive layer for fixing the ion-permeable membranes and the porous membrane on a predetermined substrate. The method according to claim 1,
Wherein the buffer reservoir and the reaction membrane are made of cellulose paper. Forming two patterned selective ion-permeable membranes to be spaced apart from one another in a straight line; And
Forming a patterned porous membrane between the selective iontophoretic membranes and one at each end,
Wherein the porous membrane formed at both ends of the selective ion-permeable membranes is a buffer reservoir, and the porous membrane formed between the selective ion-permeable membranes is a reaction membrane. 13. The method of claim 12,
Wherein the buffer reservoir and the reaction membrane are formed such that their ends partially overlap each other at a side in contact with the selective ion-permeable membranes. 13. The method of claim 12,
Wherein the selective ion-permeable membranes are attached after patterning by cutting, or formed by a technique of microflow patterning or pipetting. 13. The method of claim 12,
Wherein the buffer reservoir and the reaction membrane are formed by patterning by cutting or by patterning using a hydrophobic material. 13. The method of claim 12,
Wherein power is applied to each of the buffer reservoirs at both ends to concentrate biomolecules on the reaction membrane, and a sample containing the biomolecule is administered to the reaction membrane. 13. The method of claim 12,
Forming a channel with the porous membrane such that one side of the porous membrane is connected to a portion extending from the reaction membrane; And
Forming a sample injection membrane made of the porous membrane on the other side of the channel in a predetermined shape having a portion having a width larger than the width of the channel
Wherein the biomolecule concentrating apparatus further comprises: 18. The method of claim 17,
Forming the channel first and then forming the reaction membrane,
And a portion extending from the reaction membrane on both side ends of the channel and a part of the sample injection membrane. 18. The method of claim 17,
Wherein the reaction membrane, the channel, and the sample injection membrane are integrally formed. 18. The method of claim 17,
Wherein the buffer reservoirs at both ends of the buffer reservoir are energized to concentrate biomolecules on the reaction membrane, and a sample containing the biomolecules is administered to the sample injection membrane. . 13. The method of claim 12,
Further comprising the step of forming an adhesive layer for fixing the ion-permeable membranes and the porous membrane on a predetermined substrate, prior to the step of forming the ion-permeable membranes. 13. The method of claim 12,
Wherein the buffer reservoir and the reaction membrane are made of cellulose paper.

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