KR101710885B1 - Protein preconcentration device using capillary and fabrication method thereof - Google Patents

Abstract

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a protein concentration device and a method for manufacturing the same, and more particularly, to a protein concentration device using a fluid channel of a capillary having a patterned material capable of selectively ion- A pair of protein sample reservoirs which are formed at both ends of the capillary and in which a positive electrode and a negative electrode are connected to each other to generate a potential difference across both ends of the fluid channel, A pressure generator provided on the reservoir of the reservoir and generating pressure in the fluid channel, a substance provided in a predetermined position of the capillary, and capable of permeating ions in the protein sample moving through the fluid channel when the potential difference and the pressure are generated An ion-selective membrane selectively permeable to the capillary, Provided to a fixed micro magnetic bead extraction unit body, to extract the target substance binding to the antibody, it characterized in that it comprises a capillary tube installed on the outer circumference side is the micro magnetic beads provided for generating a magnetic force generating magnetic force.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a protein concentration apparatus using a capillary,

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a protein concentration device and a method of manufacturing the same, and more particularly, to a protein concentration device using a fluid channel of a capillary having selective ion permeable materials patterned thereon.

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. That is, from simple substances such as blood glucose for diagnosis of diabetes to genes such as bcr-abl gene fusion of chronic myelogenous leukemia, which is a therapeutic target of Gleevec, are all biomarkers, and they are practically used biomarkers 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 constituting the body of an organism, and is a substance that acts as a catalyst for various chemical reactions in cells 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. .

However, the technologies and devices for analyzing the proteins that are currently being introduced are relatively expensive and require a long time. In addition, with the technologies and devices currently introduced, it is time-consuming and expensive to perform qualitative or quantitative analysis when the amount of the sample is small, while it is difficult to concentrate when the amount of the sample increases.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a protein concentration device capable of concentrating a protein very simply and efficiently using a selective ion-permeable membrane, a capillary, and an electrode

It is also an object of the present invention to provide a protein concentration device that is excellent in concentration efficiency by forming a selective ion permeable membrane in a fluid channel.

In addition, it is possible to utilize bioassay (biosensor) and protein separation by using a simple device such as micro magnetic beads (magnetic beads) in the concentrating device itself, The present invention aims to provide a device having excellent characteristics.

In order to achieve the above object, a protein concentration device according to an embodiment of the present invention includes a capillary for forming a fluid channel, an ion selective membrane formed by providing a material capable of selectively transmitting ions at a predetermined position of the capillary, A pair of protein sample reservoirs formed at both ends of the protein sample reservoir and connected to the positive electrode and the negative electrode to generate a potential difference across both ends of the fluid channel; An extraction unit provided with a pressure generator for generating a pressure in the fluid channel, an extraction unit provided with a micro magnetic bead in which an antibody binding to a target substance is immobilized in the capillary, and a micro- And a magnetic force generating part provided on the outer surface of the capillary to generate a magnetic force The.

According to another aspect of the present invention, there is provided a method of concentrating a protein using a capillary comprising the steps of: forming an ion selective membrane through which a substance capable of selectively ion permeation is patterned at a predetermined position of a capillary forming a fluid channel; Injecting micro-magnetic beads with a binding antibody immobilized thereon; generating an electric potential difference in the capillary by connecting an external power source to a protein sample reservoir provided at both ends of the capillary to house the protein sample; Generating a pressure in the capillary by a pressure generator provided on a reservoir of one of the protein sample reservoirs; and detecting a substance in the protein sample moving in the fluid channel by the potential difference and the pressure, Transmitting an ion-selective membrane, and injecting the micro- Generates a magnetic force on the outer peripheral side of the capillary is characterized in that it comprises a step of extracting a target material bonded by the antibody from the material that has passed through the ion selective membrane.

According to the embodiment of the present invention, the protein substances to be selectively separated in the sample can be efficiently concentrated to a specific position in a very short time.

Also, by using a general-purpose capillary tube, it is very easy to fabricate the device, and the device has excellent expandability and wide application range.

1 is a view showing a configuration of a protein concentration device according to a first embodiment of the present invention.
2 is a view for explaining a method of patterning an ion selective film at a predetermined position of a capillary tube according to the first embodiment of the present invention.
FIG. 3 is a view showing a method of concentration of the protein concentration device of FIG. 1 according to the first embodiment of the present invention.
4 is a view illustrating a method of utilizing a protein concentration device according to a second embodiment of the present invention.
5 is a view illustrating a method of utilizing a protein concentration device according to a third embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

In describing the embodiments, descriptions of techniques which are well known in the technical field to which this specification belongs and which are not directly related to this specification are not described. This is for the sake of clarity without omitting the unnecessary explanation and without giving the gist of the present invention.

For the same reason, some of the components in the drawings are exaggerated, omitted, or schematically illustrated. Also, the size of each component does not entirely reflect the actual size. In the drawings, the same or corresponding components are denoted by the same reference numerals.

Hereinafter, the present specification will be described with reference to the drawings for explaining a protein concentration device using a capillary and a concentration method using the capillary according to the embodiments of the present invention.

Hereinafter, a method of manufacturing a protein concentration device according to an embodiment of the present invention will be described in detail with reference to the accompanying drawings.

First Embodiment

FIG. 1 illustrates a structure of a protein concentration device according to a first embodiment of the present invention. FIG. 2 illustrates a method of patterning an ion selective film 20 at a predetermined position of a capillary according to an embodiment of the present invention Fig.

The protein concentration device according to the first embodiment of the present invention comprises a capillary 10 for forming a channel, an ion-selective membrane 20 formed by patterning a material capable of selectively ion-permeating the inner surface of a center portion of the capillary tube, A protein sample reservoir (30, 40) formed at the end of the protein sample reservoir and connected to the positive electrode and the negative electrode to generate a potential difference across the fluid channel, an external electrode And a pressure generator, for example, a syringe 60, provided on the reservoir side of the power source 50 and the protein sample reservoir to generate pressure in the fluid channel.

A detection method for concentrating proteins using the protein concentration device shown in Fig. 1 will be described.

First, an ion selective film 20 is formed by patterning a predetermined position of a capillary tube 10 forming a fluid channel to allow selective ion permeable material to pass therethrough.

In this embodiment, naphion is used as a material capable of selective ion permeation, but a polyelectrolyte such as polystyrene sulfonate (PSS) and polyallylamine hydrochloride (PAH) may be applied. However, the present invention is not limited thereto Any material may be used as long as it is selectively ion permeable.

First, the behavior and movement of proteins generated by applying voltage to both ends of the capillary will be described.

When a sample such as blood is injected into the capillary, the surface of the capillary comes into contact with the blood fluid, and induction charges of different nature are generated between the two. A specific layer in which induced charges generated in the fluid exist is called an electric double layer (EDL). When an electric field is applied to the capillary, the ions in the fluid are attracted to the electrode opposite to the electrical properties of the ions. In this way, the ions move in the capillary 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).

2 is a diagram illustrating a method of patterning an ion selective film 20 at a predetermined position of a capillary tube according to an embodiment of the present invention.

2A is a cross-sectional view of a capillary tube used in the present embodiment.

2B, in order to form an ion selective membrane functioning as a filter in the central portion of the capillary tube 10, a needle 80 of a syringe is inserted and a selective ion permeable substance 21 such as Nafion is injected do.

The ion-selective membrane 20 is then patterned on the inner surface of the central portion of the capillary 10, as shown in FIG. 2C, by heating the capillary 10 injected with the selectively ion-permeable substance 21 at about 80.degree.

The ion selective membrane 20 serves as a kind of nanofilter that selectively transmits protons. For example, when the ion-selective membrane 20 is nafion, H + ions are selectively and rapidly permeated by the hopping and vehicle mechanism due to SO3- in the chemical structure of the ion. . Accordingly, the ion selective membrane 20 functions as a nanochannel, and protein substances to be analyzed can be efficiently concentrated in the front end toward the positive electrode of the nanochannel in a very short time.

1, the positive electrode and the negative electrode are connected to both ends of the capillary tube 10 forming the nano channel, thereby forming a protein sample reservoir the reservoirs 30 and 40 are connected. The reservoirs 30 and 40 are connected to an external power source 50 that supplies power through wires. According to another embodiment of the present invention, the reservoirs 30 and 40 may be provided with thin-film electrodes to be connected to an external power source.

In this embodiment, a capillary having an inner diameter of 1 to 2 mm is used, but it is not limited thereto. However, if the inner diameter of the capillary tube is too large, a larger pressure must be applied for the concentration, and if the capillary is applied to the capillary tube, a concentration of the capillary becomes excessively large. On the other hand, when the inner diameter of the capillary becomes smaller, the Nafion patterning becomes difficult, and it may be difficult to sufficiently obtain the effect of the pressing. The silica capillary tube 10 is filled with a buffer solution containing an electrolyte. Each end of the capillary tube 10 is positioned within the reservoir 30, 40, which is a separate fluid reservoir that receives the buffer electrolyte.

A first potential voltage is placed on one of the reservoirs (30) and a second potential voltage is placed on another reservoir (40). Thus, the positively charged particles and the negatively charged particles in the protein sample undergo the influence of the electric field formed by the two potential voltages applied to the reservoirs 30 and 40, respectively, and move through the capillary 10 in opposite directions .

On the other hand, a pressure is generated in the capillary tube 10 by the syringe 60 placed on the reservoir 30. Pressurization is for controlling the flow of fluid generated by electrophoresis and for controlling the position and concentration of the concentrating plug. Thus, by controlling the potential difference and the pressure, the moving speed, the concentration ratio, and the concentration position of the concentration plug can be controlled.

In other words, the electroosmotic flow, electrophoretic mobility and pressurization of each component of the fluid determine the overall movement for each fluid component. That is, the mobility of the fluid is the sum of the electroosmotic property, the electrophoretic mobility and the pressure, and the velocity of the fluid is the sum of the electroosmotic velocity, the electrophoretic velocity and the pressure.

On the other hand, the pressure generated in the direction of the anode from the cathode is provided on the side of the reservoir 40 where the cathode is located, and the pressure generated in the direction of the cathode from the anode is provided on the reservoir 30 side where the anode is located, Is called positive pressure.

On the other hand, the ion selective membrane 20 provided at the center of the capillary tube 10 performs a role of a nanochannel so that specific ions are selectively and rapidly transmitted.

FIG. 3 is a view showing a protein concentration method using the protein concentration device of FIG. 1;

As shown in Fig. 3, the particles selectively passing through the ion selective film 20 provided at the center of the capillary tube 10 form a concentration plug. At this time, the electroosmosis and electrophoretic mobility (F2) and the pressure (F1) described above are combined to increase the concentration ratio. It is also possible to control the position of the concentrating plug by adjusting this force.

It is also possible to directly concentrate the protein particles to be analyzed in the specific region of the capillary 10 and to install the plug in the concentration region 70 to supply concentrated protein particles to various MEMS sensors or fluidic sensors to detect the concentrated sample It is possible. In addition, the protein concentration device according to the present embodiment is advantageous in that it can be easily connected to a mass spectrometer or ESI Spray or the like, and thus can freely interoperate with various analysis equipment.

Second Embodiment

4 is a view illustrating a method of utilizing a protein concentration device according to a second embodiment of the present invention.

The protein concentration device according to the second embodiment of the present invention comprises a capillary tube 10, an ion selective membrane 20 formed by patterning a material capable of selectively ionizing the inner surface of a central portion of the capillary tube, A reservoir 30, 40, which is connected to a positive electrode and a negative electrode to generate a potential difference across both ends of the fluid channel, FIG. 4 illustrates a method of using the protein concentration device according to the second embodiment of the present invention Fig.

The protein concentration device according to the second embodiment of the present invention comprises a capillary tube 10, an ion selective membrane 20 formed by patterning a material capable of selectively ionizing the inner surface of a central portion of the capillary tube, A protein sample reservoir 30 and 40 connected to a positive electrode and a negative electrode to generate a potential difference across the fluid channel, an external power source 50 generating a potential difference between the positive electrode and the negative electrode of the protein sample reservoir, , A protein sample reservoir, and a pressure generator, for example, a syringe (60) provided on the reservoir to generate pressure in the fluid channel, a micro magnetic bead (80) to which an antibody binding to the target substance is immobilized, And a magnetic force generating portion 90 provided on the outer surface of the capillary having the magnetic beads 80 to generate a magnetic force.

The magnetic force generating section 90 is not particularly limited as long as it can apply a magnetic force, but it is preferably a permanent magnet for the sake of convenience of use.

Next, a detection method for concentrating the protein using the protein concentration device shown in Fig. 4 will be described.

First, an ion selective film 20 is formed by patterning a predetermined position of a capillary tube 10 forming a fluid channel to allow selective ion permeable material to pass therethrough.

In this embodiment, naphion is used as a material capable of selective ion permeation, but a polyelectrolyte such as polystyrene sulfonate (PSS) or polyallylamine hydrochloride (PAH) may be applied. However, the present invention is not limited thereto Any material may be used as long as it is selectively ion permeable.

2B, in order to form an ion selective membrane functioning as a filter in the center of the capillary tube 10, a needle 80 of a syringe is inserted and a selective ion permeable substance 20 such as Nafion is injected do.

The ion-selective membrane 20 is then patterned on the inner surface of the central portion of the capillary 10 by heating the capillary 10 having the selectively ion-permeable substance 20 at about 80 ° C., as shown in FIG.

The ion selective membrane 20 serves as a kind of nanofilter for selectively transmitting protons. For example, when the ion-selective membrane 20 is nafion, H + ions are selectively and rapidly permeated by the hopping and vehicle mechanism due to SO3- in the chemical structure of the ion. . Accordingly, the ion selective membrane 20 functions as a nanochannel, and protein substances to be analyzed can be efficiently concentrated in the front end toward the positive electrode of the nanochannel in a very short time.

Micro-magnetic beads having immobilized antibodies binding to the target substance are injected into the capillary in which the ion-selective membrane 20 is formed.

One example of a method for injecting micro magnetic beads is the needle needle method described in Figure 2B. In the needle needle method, a needle of a syringe containing micro-magnetic beads with an antibody fixed thereto, which binds with a target substance to be concentrated, is inserted to a desired position of the capillary tube 10, and then micro-magnetic beads are injected. In an embodiment of the present invention, the use of a needle needle method for injecting a micro magnetic bead is illustrated, but the present invention is not limited thereto.

Antibodies can be useful when it is desired to detect very low concentrations of substances or cells, since only certain substances or cells of interest can be selectively detected or isolated.

The immobilization of the antibody 84 to the micro magnetic beads 80 can be carried out using immobilization methods known in the art. For example, the antibody can be immobilized after chemically treating the surface of the micro magnetic bead particle to bind to the antibody. That is, the micro magnetic bead particle has an activating group capable of binding to the antibody, so that the antibody can be immobilized on the bead particle. In addition, the antibody can be immobilized by attaching a ligand, which binds to a specific receptor present in the antibody, to the surface of the micro magnetic bead particle and then binding the antibody.

A protein sample reservoir 30 or 40 is provided at both ends of the capillary tube 10 provided with the ion selective membrane 20 and the micro magnetic beads 80 so that the positive electrode and the negative electrode are connected to each other to generate a potential difference. Lt; / RTI > The reservoirs 30 and 40 are connected to an external power source 50 that supplies power through wires. According to another embodiment of the present invention, the reservoirs 30 and 40 may be provided with thin-film electrodes to be connected to an external power source.

As described above, capillary electrophoresis as in Fig. 2 is a technique that utilizes the electrophoretic characteristics of molecules and / or the electroosmotic flow of a sample in a small capillary to separate sample components.

According to the present embodiment, the silica capillary tube 10 is filled with a buffer solution containing an electrolyte. Each end of the capillary tube 10 is positioned within the reservoir 30, 40, which is a separate fluid reservoir that receives the buffer electrolyte.

A first potential voltage is placed on one of the reservoirs (30) and a second potential voltage is placed on another reservoir (40). Thus, the positively charged particles and the negatively charged particles in the protein sample undergo the influence of the electric field formed by the two potential voltages applied to the reservoirs 30 and 40, respectively, and move through the capillary 10 in opposite directions .

On the other hand, a pressure is generated in the capillary tube 10 by the syringe 60 placed on the reservoir 30. The generated pressure will affect the entire fluid.

At this time, the electroosmotic flow, electrophoretic mobility and pressure of each component of the fluid determine the overall movement for each fluid component. That is, the mobility of the fluid is the sum of the electroosmotic property, the electrophoretic mobility and the pressure, and the velocity of the fluid is the sum of the electroosmotic velocity, the electrophoretic velocity and the pressure.

On the other hand, the ion selective membrane 20 provided at the center of the capillary tube 10 performs a role of a nanochannel so that specific ions are selectively and rapidly transmitted.

A portion (70) of the protein particles transmitted through the ion selective membrane (20) is concentrated by the vortex formed by the electroosmosis and electrophoretic mobility (F2) and the pressure (F1) described above.

On the other hand, it is bound to the antibody 82 of the micro magnetic beads 80 among the protein particles moving out of the vortex formed by the electroosmosis and electrophoretic mobility F 2 and the pressure F 1 according to the intrinsic property of the substance.

The micro magnetic beads 80 bound to the target material are captured by the magnetic force generated by the magnetic force generating portion 90 provided on the outer peripheral side surface of the capillary tube 10 provided with the micro magnetic beads 80. [ As a result, it becomes possible to detect the substance bound to the antibody 82.

Third Embodiment

5 is a view showing a method of utilizing a protein concentration device according to a third embodiment of the present invention.

The protein concentration device according to the third embodiment of the present invention includes a capillary 10 for forming a fluid channel, an ion selective membrane 20 formed by patterning a material capable of selectively ion-permeating the inner surface of a center portion of the capillary, A protein sample reservoir (30, 40) formed at both ends and connected to the positive electrode and the negative electrode to generate a potential difference across the fluid channel, a potential difference between the positive electrode and the negative electrode of the protein sample reservoir An external power supply (50), a syringe (60) provided on one of the reservoirs of the protein sample reservoir for generating a pressure in the fluid channel, a first micro magnetism A bead 80, a second micro magnetic beads 180 to which a second antibody binding to the second target material is immobilized, and a micro magnetic beads 80, Which may comprise a magnetic force generating unit 90. The magnetic force generating section 90 may be provided in the area where the first micro magnetic beads 80 are provided and the area where the second micro magnetic beads 180 are provided as shown in FIG. In another embodiment, one magnetic force generating unit 90 may be provided to cover both the area where the first micro magnetic beads 80 are provided and the area where the second micro magnetic beads 180 are provided.

The magnetic force generating section 90 is not particularly limited as long as it can apply a magnetic force, but it is preferably a permanent magnet for the sake of convenience of use.

Next, a detection method of concentrating the protein using the protein concentration device shown in Fig. 5 will be described.

First, an ion selective film 20 is formed by patterning a predetermined position of a capillary tube 10 forming a fluid channel to allow selective ion permeable material to pass therethrough.

In this embodiment, naphion is used as a material capable of selective ion permeation, but a polyelectrolyte such as polystyrene sulfonate (PSS) or polyallylamine hydrochloride (PAH) may be applied. However, the present invention is not limited thereto Any material may be used as long as it is selectively ion permeable.

As described in the first embodiment and the second embodiment, in order to form an ion selective membrane serving as a filter in the center of the inside of the capillary tube 10, a needle capable of selectively ion-permeable, such as Nafion, (20) is injected and heated so as to be patterned on the inner surface of the capillary.

The ion-selective membrane 20 serves as a kind of nanofilter that selectively transmits a proton).

Micro-magnetic beads having immobilized antibodies binding to the target substance are injected into the capillary in which the ion-selective membrane 20 is formed.

To inject the micro magnetic beads, as described in the second embodiment, a needle needle method is used. That is, the needle of the syringe containing the micro-magnetic beads immobilized with the antibody binding to the target substance to be concentrated is inserted to the desired position of the capillary 10, and then the micro magnetic beads are injected.

That is, the needles of the syringe are inserted from one end of the capillary to the center thereof, and the first micro magnetic beads 80 to which the first antibody binding to the first target material is immobilized are firstly injected, The second micro magnetic beads 180 to which the second antibody is immobilized. In this case, the second micro magnetic beads 180 are separated from the second target material by a predetermined distance.

At this time, the second target material becomes the detection target material. The first target substance is a substance which is disturbed when detecting the second target substance, which is the substance to be detected, and is used for separating from the second target substance.

The method of immobilizing the antibody on the micro magnetic beads has been described in the second embodiment, and a description thereof will be omitted.

A positive electrode and a negative electrode are connected to both ends of the capillary tube 10 provided with the ion selective membrane 20, the first minute magnetic beads 80 and the second minute magnetic beads 180, Connect sample reservoirs (30, 40). The reservoirs 30 and 40 are connected to an external power source 50 that supplies power through wires. According to another embodiment of the present invention, the reservoirs 30 and 40 may be provided with thin-film electrodes to be connected to an external power source. Such an electrode is merely an example, but is not limited thereto.

As described above, capillary electrophoresis as in Fig. 2 is a technique that utilizes the electrophoretic characteristics of molecules and / or the electroosmotic flow of a sample in a small capillary to separate sample components.

However, the capillary 10 allows the ion-selective membrane 20 to be in the negative electrode direction with respect to the region where the micro magnetic beads 80 and 180 are implanted.

According to the present embodiment, the silica capillary tube 10 is filled with a buffer solution containing an electrolyte. Each end of the capillary tube 10 is positioned within the reservoir 30, 40, which is a separate fluid reservoir that receives the buffer electrolyte.

A second potential voltage is placed on the first potential voltage reservoir (40) in one of the reservoirs (30). Thus, the positively charged particles and the negatively charged particles in the protein sample undergo the influence of the electric field formed by the different potential voltages applied to the reservoirs 30, 40 and move in opposite directions through the capillary tube 10, respectively . Further, a pressure is generated in the capillary tube 10 by the injector 60.

At this time, the electroosmotic flow, electrophoretic mobility and pressure of each component of the fluid determine the overall movement for each fluid component. The potential difference and the pressure are adjusted to adjust the moving speed, the concentration ratio, and the concentration position of the concentration plug.

On the other hand, the ion selective membrane 20 provided at the center of the capillary tube 10 performs a role of a nanochannel so that specific ions are selectively and rapidly transmitted.

A portion (70) of the protein particles transmitted through the ion selective membrane (20) is concentrated by the vortex formed by the electroosmosis and electrophoretic mobility (F2) and the pressure (F1) described above.

On the other hand, binding to the antibody 82 of the first minute magnetic beads 80 among the protein particles moving out of the vortex formed by the electroosmosis and electrophoretic mobility F2 and the pressure F1 according to the intrinsic properties of the substance do.

The first minute magnetic beads 80 combined with the target material are captured by the magnetic force generated by the magnetic force generating portion 92 provided on the outer peripheral side surface of the capillary tube 10 provided with the minute magnetic beads 80. [

Of the protein particles moving out of the vortex formed by electroosmosis and electrophoretic mobility (F2) and pressure (F1) depending on the intrinsic properties of the material, the second target material is the second antibody of the second micro magnetic beads 180 Lt; / RTI >

The second micro magnetic beads 180 combined with the second target material are captured by the magnetic force generated by the magnetic force generating portion 94 provided on the outer peripheral side of the capillary tube 10 provided with the second micro magnetic beads 180 .

The moving speed of the protein sample in the fluid can be controlled by controlling the intensity or the potential difference of the pressure of the syringe.

It will be understood by those skilled in the art that the present specification may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. The scope of the present specification is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present specification Should be interpreted.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, It is not intended to limit the scope of the specification. It will be apparent to those skilled in the art that other modifications based on the technical idea of the present invention are possible in addition to the embodiments disclosed herein.

10: capillary
20: ion selective membrane
30: Protein sample reservoir
50: External power source
60: Syringe
80: Micro magnetic beads
90:

Claims (9)

A capillary forming a fluid channel;
An ion selective membrane formed by providing a material capable of selectively transmitting ions at a predetermined position of the capillary;
A pair of protein sample reservoirs formed at both ends of the capillary and having positive and negative electrodes connected to each other to generate a potential difference at both ends of the fluid channel, the potential difference being adjustable;
A pressure generator provided on a reservoir side of one of the protein sample reservoirs for generating a pressure in the fluid channel, the pressure intensity being adjustable;
An extraction unit provided with a micro magnetic bead in which an antibody binding to a target substance is immobilized in the capillary; And
In order to extract a target substance binding to the antibody, a magnetic force generating unit, which is provided on the outer peripheral side of the capillary having the micro magnetic beads and generates magnetic force,
Lt; / RTI >
And adjusting the movement speed and position of the concentrating plug moving in the fluid channel by adjusting the potential difference from the external electrode and the pressure of the pressure generator. [2] The method of claim 1, wherein the extracting unit comprises: a first micro magnetic bead in which a first antibody binding to a first target substance is immobilized, and a second micro magnetic bead in which a second antibody binding to a second target substance is immobilized, And a capillary-type protein concentration device. 3. The method according to claim 1 or 2,
Wherein the micro magnetic beads are inserted from one end of the capillary to the center and injected with the micro magnetic beads. The capillary-based protein concentration device according to claim 1, wherein predetermined protein particles in the protein sample are concentrated at the front end of the ion selective membrane while passing through the fluid channel in which a potential difference and a pressure are generated. delete The method according to claim 1,
Wherein the selective ion permeable material is nafion. ≪ RTI ID = 0.0 > 11. < / RTI > Forming an ion selective film through a selective ion permeable substance at a predetermined position of a capillary forming a fluid channel;
Injecting a micro magnetic bead having an antibody binding to the target substance immobilized therein;
A capillary disposed at both ends of the capillary and connected to an external power source through a pair of electrodes provided in a protein sample reservoir for receiving a protein sample to generate a potential difference between the capillary and one of the protein sample reservoirs, Generating a pressure in the capillary by a pressure generator provided on a side of the capillary;
Transmitting only the ion permeable substance in the protein sample moving in the fluid channel by the generation of the potential difference and the pressure through the ion selective membrane;
Generating a magnetic force on an outer peripheral side surface of the capillary into which the micro magnetic beads are injected to extract a target substance bound by the antibody among the substances transmitted through the ion selective membrane,
And controlling the moving speed and position of the concentrating plug moving in the fluid channel by adjusting the potential difference from the external electrode and the pressure intensity of the pressure generator in the step of extracting the combined target material A method of protein concentration using a capillary. 8. The method of claim 7,
In the step of injecting the micro magnetic beads to which the antibody is immobilized,
A first micro magnetic bead having a first antibody immobilized thereon immobilized on a first target substance is first injected from the one end of the capillary to the center and a second micro magnetic bead with a second antibody immobilized thereon, Wherein the first micro magnetic beads are injected at a distance from the first micro magnetic beads.
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