KR20200074832A - Sample Concentration and Separation Device by Controlling Speed and Position of Sample in Microchannel - Google Patents

Abstract

The present embodiments provide a sample concentrating and separating apparatus. The sample concentrating and separating apparatus of the present invention attaches an ion exchange membrane to a channel having micropores along the moving direction of an analyte and controls a movement speed of the analyte, and a detection unit is positioned near the ion exchange membrane. Therefore, the analyte is highly concentrated and separated even without application of an electric field. The sample concentrating and separating apparatus includes the channel and the ion exchange membrane.

Description

{Sample Concentration and Separation Device by Controlling Speed and Position of Sample in Microchannel}

The technical field to which the present invention pertains relates to a sample concentration and separation device.

The contents described in this section merely provide background information for this embodiment, and do not constitute a prior art.

Modern medicine aims not to simply extend the lifespan, but also to extend the lifespan of healthy and long-lived health. Therefore, the paradigm is changing, as the future medicine is not centered on therapeutic medicine, but is implementing 3P of preventive medicine, predictive medicine, and personalized medicine. In order to realize this in detail, early detection and early treatment of diseases have become very important means, and as a means for this, research on biomarkers has been actively conducted.

Biomarkers are markers that can distinguish normal or pathological conditions, predict treatment responses, and measure objectively. In biomarkers, nucleic acids (DNA, RNA), proteins, fats, metabolites, and changes in their patterns are used. That is, from simple substances such as blood glucose for the diagnosis of diabetes to genes such as BCR-ABL gene fusion of chronic myelogenous leukemia, which is a therapeutic target of Gleevec, all are biomarkers and are biomarkers that are actually used in clinical practice.

Analysis of nucleic acids or proteins can determine the degree of disease expression and progression. Techniques and devices for protein analysis have a problem in that it is difficult to manufacture devices by using nanotechnology and are relatively expensive to be widely used. In addition, there is a disadvantage in that a high-sensitivity sensor is required in a protein analysis device, or accurate analysis is difficult with a small sample. A typical method for detecting nucleic acids or proteins is a lateral flow analysis method using a chromatography method. This lateral flow analysis method is used in various fields such as pregnancy diagnosis.

Pregnancy diagnostic kits use monoclonal antibodies that bind chorionic gonadotropin (HCG). A chromogen is attached to the HCG antibody, which is a monoclonal antibody that responds only to HCG. The HCG in the urine moves after binding with the HCG antibody of the pregnancy diagnostic kit, and then the HCG complex and the antibody that binds to HCG bind in the pregnancy display window, and a red band appears. Antibodies that bind to bind red bands.

HCG usually has a high concentration, so it is easy to detect, but other biomarkers have a low concentration, which makes it difficult to detect.

1 is a diagram illustrating a lateral flow analysis strip to which the ion exchange membrane 124 is attached to the receiving portion 120. Concentration kit 122 is attached to the receiving portion 120. When an electric field is applied through the power source 126 to the ion exchange membrane attached to the receiving portion, a specific analyte is concentrated at an intermediate point of the receiving portion 120. An electric field must be applied to concentrate the analyte at a high concentration through the ion exchange membrane attached to the receiving portion shown in FIG. 1.

Korean Open Patent No. 10-2018-0056342 (published May 28, 2018)

Embodiments of the present invention, by attaching the ion exchange membrane along the direction of movement of the analyte in a channel having micropores, the main object of the invention is to concentrate the analyte nearly 10 times or more and separate the analyte without applying an electric field. have.

Other unspecified objects of the present invention may be further considered within a range that can be easily deduced from the following detailed description and its effects.

According to an aspect of the present embodiment, a sample concentration and separation device for measuring an analyte in a sample, comprising: a channel for moving the analyte with fine pores; And an ion exchange membrane attached to the channel and having a predetermined length along a direction in which the analyte moves.

The sample concentration and separation device may be located at a predetermined distance from the ion exchange membrane and attached to the channel, and may include a detection unit that detects an analyte concentrated by the ion exchange membrane.

A depletion force and a depletion region are formed in a region where the ion exchange membrane is in contact with the channel to reduce a cross-sectional area of the channel through which the analyte will pass, and the analyte moving speed can be reduced over time.

The phenomenon in which the movement speed of the analyte is delayed while passing through the micropores of the channel and the depletion force caused by the ion exchange membrane attached to the channel interact, perpendicular to the position where the ion exchange membrane is attached. The analyte can be concentrated at one point or at a region adjacent to the vertical point.

The size of the micropores in the channel to which the ion exchange membrane is attached is reduced, so that the first analyte having a size smaller than the size of the reduced micropores passes through and the agent having a size larger than the size of the reduced micropores 2 As the analyte does not pass, the analyte may be separated.

The ion exchange membrane is arranged to have a predetermined length along the direction in which the channel extends, and the length, thickness, width, or combination thereof can be adjusted to control the size of the depletion force and depletion region. .

The location in which the analyte is concentrated in the channel may be changed depending on the presence or absence of a surfactant in the sample.

The hydrogen ion concentration (pH) of the analyte moving the channel may be adjusted within a predetermined range using a buffer solution having a hydrogen ion concentration set according to the polarity of the ion exchange membrane.

The ion exchange membrane includes both a cation exchange membrane (CEM) and an anion exchange membrane (AEM), and an analyte moving the channel by contacting or spaced apart the cation exchange membrane and the anion exchange membrane The hydrogen ion concentration (pH) can be adjusted within a predetermined range.

It may be connected to the channel, and may include a conjugate portion including a conjugate that combines a detector and an indicator to form a complex by binding with the analyte.

The detection unit may include a first capturer that captures the complex.

The channel may include a control portion including the conjugate that does not form the complex or the second trapper that captures the detector.

The ion exchange membrane may be attached over the conjugate portion.

The ion exchange membrane can be attached to the top, bottom, side, or combined positions of the channels.

The number of ion exchange membranes is plural, and the plurality of ion exchange membranes may be arranged in a zigzag manner or formed in a stacked structure.

A concentration position controller may be positioned between the ion exchange membrane and the channel.

The channel may include (i) a straight section, (ii) a curved section, (iii) a'v' shape or a curved section including a'a' shape, (iv) a slope section with different heights, and (v) a'X' shape. Intersecting section comprising, (vi) a branching section containing a'Y' shape, (vii) a stacking section in which a plurality of channels are layered, (viii) a through section in which upper and lower channels are connected via vias, or a combination thereof. It can contain.

The sample may further include an inlet that is a passage through which the sample flows and an outlet that is a passage through which the sample flows.

It may be further connected to the channel receiving portion having a space for receiving the sample.

Connected to the channel, it may further include an absorber for absorbing the sample by capillary action.

The sample concentration and separation device may be implemented as a lateral flow array or microfluidic chip.

As described above, according to embodiments of the present invention, by attaching the ion exchange membrane along the direction of movement of the analyte to a channel having micropores, and placing the ion exchange membrane on the detection portion formed in the channel, an analyte without applying an electric field It has the effect of being highly concentrated and separable.

Even the effects not explicitly mentioned herein, the effects described in the following specification expected by the technical features of the present invention and the potential effects thereof are treated as described in the present specification.

1 is a view illustrating an lateral flow analysis strip having an ion exchange membrane attached to a receiving portion.
2A to 2F are views illustrating a structure in which an ion exchange membrane is attached to a channel in a sample concentration and separation device according to embodiments of the present invention.
3 to 10 are diagrams illustrating a sample concentration and separation device according to embodiments of the present invention.
11 and 12 are diagrams illustrating a principle in which an analyte is concentrated and moved in a sample concentration and separation device according to embodiments of the present invention.
13A, 13B, 14, and 15 are diagrams illustrating the results of concentrating an analyte using a sample concentration and separation device according to embodiments of the present invention.

Hereinafter, when it is determined that the subject matter of the present invention may be unnecessarily obscured by those skilled in the art with respect to known functions related to the present invention, the detailed description thereof will be omitted, and some embodiments of the present invention will be described. It will be described in detail through exemplary drawings.

The sample concentration and separation device according to the present embodiment may be implemented as a structure including a channel and an ion exchange membrane, and may be commercialized as a lateral flow array or a microfluidic chip. The sample concentration and separation device moves a sample or an analyte in a sample using a physical force such as pressure or gravity, mechanical or electrical power when the sample enters the microchannel.

The sample concentration and separation device includes a channel, an ion exchange membrane, and a detection unit. The channel can be connected to the support. The sample concentration and separation device may further include a protective case or the like. The ion exchange membrane is attached to the channel. Both a cation exchange membrane and an anion exchange membrane can be attached to the channel.

The detection unit is located at a predetermined distance from the ion exchange membrane, is attached to the channel, and detects the analyte concentrated by the ion exchange membrane. The detector may be attached to the channel spaced apart from the position where the ion exchange membrane is attached. The ion exchange membrane and the detection unit may be attached at positions opposite to each other based on the channel. The sample concentration and separation device adjusts the position of the concentration point or concentration area of the analyte with an ion exchange membrane, and positions the detection unit at the concentration point or concentration area.

2A to 2F are diagrams illustrating a lateral flow analysis strip in which a sample concentration and separation device having an ion exchange membrane attached to a channel according to embodiments of the present invention is implemented.

The channel includes (i) a straight section, (ii) a curved section, (iii) a'v' shape or a bent section comprising a'a' shape, (iv) a slope section with different heights, and (v) a'X' shape. Intersecting section, (vi) a branch section including a'Y' shape, (vii) a stacked section in which a plurality of channels are layered, (viii) a through section in which upper and lower channels are connected via vias, or a combination thereof can do.

Referring to FIG. 2A, the sample concentration and separation device may include a channel 100, an ion exchange membrane 200, and a detector 140.

One end of the channel 100 may include an inlet 101, and the other end of the channel 100 may include an outlet 102. The inlet portion 10 is a passage through which the sample flows, and the outlet portion 102 is a passage through which the sample flows. The fluid moves from the inlet 10 to the outlet 102. The inlet portion 101 may be formed in a funnel shape.

Referring to FIG. 2B, the sample concentration and separation device may include a receiving part 120, a conjugate part 130, a channel 100, an ion exchange membrane 200, and an absorbing part 170. The inlet portion 101 may be formed in the receiving portion 120, and the outlet portion 102 may be formed in the absorbing portion 170.

The receiving part 120, the conjugate part 130, the channel 100, and the absorbing part 160 may be fixed to the support 110, and the channel 160 may serve as the support 110.

The receiving unit 120 serves as a buffer for receiving a sample to be detected. The conjugate portion 130 includes a detector coupled to the chromogenic material. The detector binds to the analyte in the sample. The detector is separated from the conjugate portion 130 as the sample moves. The channel 100 includes a detection unit 140 and a control unit 150, indicates whether an analyte is present in the sample, and indicates whether the sample is moving. The absorber 160 is a space that absorbs a sample using a capillary phenomenon.

The conjugate unit 130 is connected to the channel 100 and includes a combination of a detector and an indicator that combine with an analyte to form a complex. The indicator is a substance that generates a signal for detection using the naked eye or a sensor. The indicator may be a coloring material, for example, gold nanoparticles, but is not limited thereto, and a suitable material may be used depending on the design to be implemented.

The channel 100 includes at least one of the detection unit 140 and the control unit 150. The detection unit 140 detects whether an analyte is present in the sample by capturing the analyte directly or indirectly. The detection unit 140 includes a first capturer that captures an analyte or complex. The control unit 150 includes a second trapper that captures a conjugate or a detector that does not form a complex. That is, the control unit 150 checks whether or not the sample has moved regardless of the presence or absence of an analyte.

In the conjugate portion, an indicator-detector conjugate is located, and as the analyte passes through the conjugate portion, an indicator-detector-analyte complex is formed. The shifted indicator-detector-analyte complex is captured by the first capturer at the detection site.

The reaction of forming a complex in the conjugate portion or capturing in a channel may be formed by a physical reaction, a chemical reaction, a biological reaction, or a combination thereof. For example, it may be an antigen-antibody reaction.

The method of detecting a sample or analyte in a sample concentration and separation device is not limited to an antibody-antigen reaction, and the binding sites (ligands) referred to herein are protein ligands (Ligand), nucleic acids (DNA) for various analytes. Or RNA) contains the binding site of the molecular sequence, and specific binding substances are proteins capable of selectively and specifically binding to the binding site, viral phage, nucleic acid molecule aptamer, hapten, DNP, etc. It includes all of the biomolecules containing, and is not limited to the description.

The path of movement of the sample comprising the channel can be implemented with a porous medium that responds to the capillary action. Media include, but are not limited to, Cellulose, Nitrocellulose, polyethersulfone, polyvinylidene, fluoride, nylon, and polytetrafluoroethylene that can be used for lateral flow. The medium can be used alone or in combination with other materials.

The ion exchange membrane 200 is attached to the channel 100. The form in which the ion exchange membrane 200 is attached to the channel 100 may be attached to the top, bottom, or side surfaces of the channel 100, or a combination thereof. Since the ion exchange membrane 200 forms a depletion force and a depletion region in a region contacting the channel, which is a movement passage, the ion exchange membrane 200 is disposed long along the channel. Mainly, the length of the ion exchange membrane 200 is controlled, and the thickness and width as well as the length are controlled to control the depletion force and depletion region. As the analyte passes through the bottom of the channel region to which the ion exchange membrane 200 is attached, the analyte is concentrated.

Due to the size of the micro pores in the channel, the movement speed decreases depending on the size and weight of the analyte, and the anion exchange membrane 200 forms a depletion force and a depletion region in the region contacting the channel, which is a movement passage, so that the analyte passes The cross-sectional area of the channel is further reduced. After a certain period of time, the analyte has a movement speed close to O at the bottom of the channel region where the ion exchange membrane 200 is attached.

Referring to FIG. 2C, the ion exchange membrane 200 may be attached over a portion of another passage, such as the conjugate portion 130.

2D and 2E, a plurality of ion exchange membranes 200, 201, and 202 may be attached. The plurality of ion exchange membranes 200, 201, and 202 may be arranged in a zigzag or formed in a stacked structure.

Referring to FIG. 2F, a concentration position adjusting unit 250 may be positioned between the ion exchange membrane 200 and the channel 100. The depletion force and depletion region of the ion exchange membrane are adjusted according to the size and position of the concentration position controller 250 to change the point at which the analyte is concentrated. That is, the analyte can be concentrated near the detector.

The location of the analyte concentration in the channel changes depending on the presence or absence of surfactant in the sample. The direction in which the movement started based on the moving direction of the analyte is called forward, and the direction in which the movement proceeds is called the back. If there is no surfactant in the sample or channel, the analyte is concentrated at a point slightly ahead of the ion exchange membrane, and if there is a surfactant, the analyte is concentrated at a point midway or slightly behind the ion exchange membrane relative to the direction of the analyte. Since the surfactant suppresses the occurrence of Ion Concentration Polarization (ICP), the higher the concentration of the surfactant, the higher the concentration point is pushed back. That is, the concentration point of the analyte can be adjusted according to the concentration of the surfactant.

The ion exchange membrane 200 may be formed to have a thickness of several hundred nanometers to tens of micrometers, and may have a width of tens to hundreds of micrometers. The length of the ion exchange membrane 200 may be formed to have tens of millimeters along the path of movement of the sample.

The ion exchange membrane 200 is also called a selective permeable membrane. Ion exchange membrane (Ion exchange membrane, IEM) is a cation exchange membrane (Cation Exchange Membrane, CEM), anion exchange membrane (Anion Exchange Membrane, AEM).

The selective ion permeable material forming the ion exchange membrane 200 refers to a material that binds well with specific ions and attracts each other, but does not bind well with other ions and does not attract. The selective ion permeable material can be, for example, nafion.

In addition to Nafion, the ion exchange membrane 200 may also be implemented as a polymer electrolyte such as polystyrene sulfonate (PSS) or polyallylamine hydrochloride (PAH), and optionally, any material capable of ion permeation may be used. Do.

The ion exchange membrane may have a form in which an ion exchange group is introduced into a fluorine-based polymer. For example, DuPont's Nafion, Dow Chemicals' Dow membrane, Asahi Chemicals' Asciplex-S membrane, Asahi Glass Glass)'s Flemion film.

On the other hand, there are ion exchange membranes developed using non-fluorine polymers and partially substituted fluorine polymers, sulfonated poly(phenylene oxide)-based, poly(phenylene sulfide)-based, polysulfone-based, and poly(para-phenylene). And ion exchange membranes using polymers such as ), polyetheretherketone, and polyimide.

Ion exchange membranes can be produced by immersing a base with fine pores in a selective ion permeation solution (resin). In view of the molecular size of the analyte, the size diameter of the micropores can range from tens to hundreds of nanometers. For example, it may be set to 200 nm or less.

3 to 10 are diagrams illustrating a sample concentration and separation device according to embodiments of the present invention.

The base includes a sample base and a passage base. The sample base stores the sample together with the receiving portion. The passage base acts like a channel. The coating layer is located in at least a portion of the base to prevent adsorption of the sample to be treated and to separate the storage space or the movement path. The sample concentration and separation device can fix the base and the ion exchange membrane with an adhesive tape or the like.

As the hydrophobic material, a wax or the like can be used as an alcohol fatty acid ester. The wax can be printed on the base or combined by heating. However, it is not limited to this, and acrylic, olefin, amide, imide, styrene, carbonate, vinyl acetal, diene, vinyl, ester, vinyl ester, ketone, fluorocarbon or Teflon ), PDMS, silane (Silane), and the like. In addition, there is an alkylsilane-based Siliclad, and the silicone-based is a hydride-terminated polydimethylsiloxane (Methylhydrosiloxane-Dimethysiloxane Copolymer).

In general, when a material becomes hydrophobic in a material, it may have hydrophobicity due to the influence of the structure of the surface in contact with water and the properties of the surface itself of the material. Particularly, the micro-channel is formed by coating the paper as the base, and the latter is applied, and forming the channel by coating the wax on the paper makes the other parts hydrophobic except for the part to be used as a channel in the hydrophilic material called paper. can do. This hydrophobic property can achieve a similar effect when using a base material similar to paper, so it is also possible to use a fiber-based base in addition to cellulose paper.

This embodiment discloses a method of coating a pattern of a hydrophobic material on paper as a method of bonding a hydrophobic material, wax, to the base. There is no particular limitation on the wax patterning method, and it is also possible to simply bond the hydrophobic material with the paper, or to bond the paper with the hydrophobic material infiltrated, but preferably, the hydrophobic material is applied to the paper to prevent leakage of the sample. It is desirable to bond to penetrate. In the case of paper, it is not only inexpensive, but also has good elasticity and moldability, and it is easy to form a coating layer having a certain thickness on the paper in the form of adsorption and penetration with a hydrophobic material, and has excellent bonding strength with a hydrophobic coating material.

The base may include a porous membrane, and the resolution may be controlled by controlling the thickness of the base to separate and concentrate the sample. For example, if the base is paper, resolution can be adjusted by differentially applying the thickness of the paper to 50 μm, 180 μm, 350 μm, etc., and the cross-sectional area of the channel can be efficiently adjusted by adjusting the size of the pores in the base. Can be controlled.

Referring to FIG. 7, in the concentration method using an ion exchange membrane (IEM) attached to a channel, a change in hydrogen ion concentration (pH) occurs at each moving point of the analyte due to ion absorption of the ion exchange membrane. Immunoassay-based lateral flow assay strips vary in response to changes in pH. The change in pH causes errors such as detection performance and false positives in the lateral flow analysis strip.

Referring to FIG. 8, a buffer solution having a polarity complementary to that of the ion exchange membrane is used to adjust the hydrogen ion concentration (pH) of the analyte moving the channel within a predetermined range. For example, when a cation exchange membrane (CEM) is attached, a change in pH can be reduced by utilizing a basic buffer solution. As a buffer solution, PBS (Phosphate-buffered saline), borate, or the like can be used. After the analyte passes through the bottom of the ion exchange membrane, the hydrogen ion concentration can be adjusted within a certain range.

Referring to FIGS. 7 and 8, when only one cation exchange membrane is attached, strong acidity appears at the bottom of the cation exchange membrane that absorbs hydrogen ions, and analytes migrate to show acidity at the back of the cation exchange membrane. That is, a difference in hydrogen ion concentration occurs before and after the analyte passes through the bottom of the ion exchange membrane.

9 and 10, the cation exchange membrane (Cation Exchange Membrane, CEM) and the anion exchange membrane (Anion Exchange Membrane, AEM) are complementarily coupled. The ion exchange membrane may include both a cation exchange membrane (CEM) and an anion exchange membrane (AEM). By placing the cation exchange membrane and the anion exchange membrane in contact or spaced apart, the hydrogen ion concentration (pH) of the analyte moving the channel can be adjusted within a predetermined range. The difference in hydrogen ion concentration can be reduced before and after the analyte passes through the bottom of the ion exchange membrane.

11 and 12 are diagrams illustrating a principle in which an analyte is concentrated and moved in a sample concentration and separation device according to embodiments of the present invention.

Ion exchange membrane (IEM) is attached to a porous material (eg, paper, cotton, thread, etc.) made of a fiber network structure, and when the sample passes through an area perpendicular to the region where the ion exchange membrane is attached, the porous material The sample is concentrated at the bottom of the ion exchange membrane in the region of.

The point where the movement speed of the sample is delayed while passing through the fine pores of the channel of the porous material, and the depletion force caused by the ion exchange membrane attached to the channel interacts, where it is perpendicular to the location where the ion exchange membrane is attached. Alternatively, the channel is moved as the analyte is concentrated in a region adjacent to the vertical point.

Depletion force refers to a force that occurs when the distance between colloids to be energized by the Short Range Attraction is sufficiently close, and does not act when it is farther than that distance. The sufficient distance here is equal to the size of another colloid, Depletant, which is smaller than a colloid, but distributed within the same system to generate a free-moving depletion force. The term depletion refers to particles that induce a depletion force. The large amount of colloidal particles bind to each other.

When the sample passes through the bottom of the ion exchange membrane due to the capillary phenomenon, only the anion or cation is selectively absorbed by the native charge of the ion exchange membrane and depletion force is generated and depletion force occurs. A depletion zone is formed.

The depletion zone decreases the width or height of a path through which a sample having charge can pass, thereby reducing sample mobility and stacking in a specific area. That is, the depletion force acts in the opposite direction of the channel to which the ion exchange membrane is attached, thereby reducing the channel passage size.

A sample having a concentration lower than the initial concentration flows out of the sample passing through the region to which the ion exchange membrane is attached and exiting the terminal.

It is advantageous in terms of portability and cost because it can improve the detection performance with no power supply by combining an ion exchange membrane with a paper-based channel.

It can also be used to separate particles of different sizes or masses. Separation of specific particles according to the mobility of particles by controlling the pore size (Pore Size) of the material in the form of a fiber network and the intensity of the depletion force and the width and height of the depletion region by the original charge of the ion exchange membrane (IEM) can do.

When simultaneously moving the first sample and the second sample having a smaller molecular size than the first sample in the channel, at the bottom of the specific point (D ₂ ) to which the ion exchange membrane having a predetermined length (D ₁ ) is attached along the direction of movement Due to the width (W ₂ ) or height (H ₂ ) of the depletion region due to the depletion force, the movable width (W ₃ ) or height (H ₃ ) is greater than the width (W ₁ ) or height (H ₁ ) of the channel. Will decrease. The first sample having a size larger than the movable width W ₃ or height H ₃ is concentrated, and the second sample having a size smaller than the movable width W ₃ or height H ₃ passes. That is, the pore size of the channel, the force to be moved (eg, capillary phenomenon), and the depletion region due to the depletion force of the ion exchange membrane (IEM) interact to play the role of a filter.

As a result, when alexa 350 (size: 1 nm or less) and albumin-FITC (size: ~ 20 nm) are concentrated together with cellulose paper, alexa 350 does not collect, and only lbumin-FITC can be observed at the bottom of the ion exchange membrane.

The method of moving the concentrated analyte after concentration of the analyte by applying an electric field to the ion exchange membrane coupled to the inlet or the receiving portion, when a voltage difference occurs, ions present in the fluid by the electric field according to the voltage difference It is drawn toward the electrode opposite to the electrical properties of the ions. In this way, ions move in the channel according to the electrical properties, leading to bringing together fluid particles by viscous force. The whole fluid flow 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 are changed near the channel implemented by the ion exchange membrane, 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 channel. At this time, the depletion zone acts as a kind of electrical barrier for an analyte having a charge due to a low ion concentration and a high electric field. As a result, the analyte does not pass through the deficient region and is concentrated before it. The analyte is concentrated very quickly in front of the depletion region in the channel. The size of the depletion region due to the ion concentration polarization expands as the ion concentration polarization of the sample progresses, so that the analyte forms a concentrated region in the center region of the channel.

On the other hand, the concentration method using the ion exchange membrane coupled to the channel according to the present embodiment reduces the cross-sectional area of the channel by forming a depletion region by a depletion force in the channel having fine pores without applying an electric field to the ion exchange membrane. There is an effect that the analyte can be concentrated several times or more, and analytes having different sizes can be filtered by reducing the moving speed of the analyte through the reduced channel.

In the channel, analytes having a size smaller than the reduced pores may be passed, and progression of analytes having a size larger than the reduced pores may be prevented and concentrated. The size of the fine pores in the channel to which the ion exchange membrane is attached is reduced, so that the first analyte having a size smaller than the size of the reduced fine pores passes through, and the second analyte having a size larger than the size of the reduced fine pores Failure to pass, the analyte is separated.

Referring to FIG. 12, a detection unit 140 is attached to the channel 100. When the first sample is moved in the channel, the width (W ₂ ) of the depletion region due to the depletion force at the bottom of the specific point (D ₂ ) to which the ion exchange membrane having a predetermined length (D ₁ ) is attached along the moving direction ) Or height (H ₂ ), the width (W ₅ ) or height (H ₅ ) of the channel is reduced than the width (W ₁ ) or height (H ₁ ) of the channel. The first sample having a size larger than the movable width W ₅ or height H ₅ is concentrated. That is, the pore size of the channel, the force to be moved (eg, capillary phenomenon), and the depletion region due to the depletion force of the ion exchange membrane (IEM) interact to play the role of a filter.

The method of sample concentration and separation involves attaching an ion exchange membrane to the channel. The method of sample concentration and separation includes injecting a sample into an inlet or a receiver. The method of sample concentration and separation includes concentrating an analyte while the analyte passes through an inlet or a receiver and passes through an ion exchange membrane-attached channel. The analyte is detected at the detector and the sample is transferred to the outlet or absorber.

In the step of concentrating the analyte, a phenomenon in which the movement speed of the analyte is delayed while passing through the micropores of the channel and a depletion force caused by the ion exchange membrane attached to the channel interacts, thereby attaching the ion exchange membrane. The analyte is concentrated at a point perpendicular to the location.

The step of concentrating the analyte forms a depletion force and depletion region in the region where the ion exchange membrane abuts the channel, reducing the cross-sectional area of the channel through which the analyte will pass, and decreasing the rate of analyte movement over time.

13A, 13B, 14, and 15 are diagrams illustrating the results of concentrating an analyte using a sample concentration and separation device according to embodiments of the present invention.

When performing the experiment, the operation time is 1800 sec, the operation volume is 100 uL, the absorbent pad is LFA (Lateral Flow Assay) Absorbent Paper, and the sample is albumin-FITC. The receiving part is GE Healthcare Chromatography paper, and the ion exchange membrane is Nafion infiltrated paper. Nafion infiltrated paper is formed by immersing Chromatography paper in 20 wt% of Nafion resin. Pad material is Chromatography paper, Nafion dilution ratio is Nafion resin 20wt% stock solution, and deposition capacity is 50-60 uL/cm ² . The infiltration conditions were dropped in the form of droplets, left for about 5 minutes, and the drying conditions were heat-treated at 80°C and 10 min. The passage base is Nitrocellulose Membrane and has a pore size of 0.2 um.

13A is a result of experiments by implementing Nitrocellulose shown in FIGS. 3 and 4 as a base, and FIG. 13B is a result of experiments by implementing cellulose shown in FIGS. 5 and 6 as a base. 13A and 13B, it can be confirmed that the analyte can be concentrated 10 times or more without using a power source.

14 and 15 are views illustrating a result of concentrating an analyte by forming a sample concentration and separation device according to embodiments of the present invention in a lateral flow analysis strip structure.

When performing the experiment in FIG. 14, the operation time is 5 min, the sample volume is 50 uL, the sample is hCG, and the buffer solution is deionized water. It can be seen that the concentration ratio of hCG increases.

In (a) of FIG. 15, when the ion exchange membrane is not attached (A), when the positions of the ion exchange membranes are set differently (B, D, F), when the cation exchange membrane and the anion exchange membrane are combined (C, E), The detection performance of Type B was evaluated using the influenza diagnostic strip.

In FIG. 15(b), it can be understood that the detection performance is improved because the gray scale is low in the ion exchange membranes (E) and (F) attached to the Type B detector.

In these embodiments, a depletion region by a depletion force is formed in a channel without applying an electric field to the ion exchange membrane to the biomarker, which is an analyte, to reduce the cross-sectional area of the channel and increase the analyte movement speed through the reduced channel. By reducing, the analyte can be separated while the analyte is concentrated several times or more.

In these embodiments, by attaching both the cation exchange membrane and the anion exchange membrane to the channel, there is an effect that the hydrogen ion concentration (pH) of the analyte moving the channel can be adjusted within a predetermined range.

The present embodiments are for explaining the technical spirit of the present embodiment, and the scope of the technical spirit of the present embodiment is not limited by these examples. The protection scope of the present embodiment should be interpreted by the claims below, and all technical spirits within the equivalent range should be interpreted as being included in the scope of the present embodiment.

10: Sample concentration and separation device
100: channel 200: ion exchange membrane
110: support 120: accommodation
130: conjugate unit 140: detection unit
150: control unit 160: absorption unit

Claims (20)

A sample concentration and separation device for measuring an analyte in a sample,
A channel having fine pores and moving the analyte; And
Ion exchange membrane attached to the channel and having a predetermined length along the direction in which the analyte moves
Sample concentration and separation device comprising a. According to claim 1,
A sample concentration and separation device comprising a detection unit that is located at a predetermined distance from the ion exchange membrane and is attached to the channel and detects an analyte concentrated by the ion exchange membrane. According to claim 1,
A depletion force and a depletion zone are formed in a region where the ion exchange membrane abuts the channel to reduce the cross-sectional area of the channel through which the analyte passes, and over time, the analyte Sample concentration and separation device, characterized by reducing the rate of movement. According to claim 1,
The phenomenon in which the movement speed of the analyte is delayed while passing through the fine pores of the channel and the depletion force by the ion exchange membrane attached to the channel interact, so that the point perpendicular to the position where the ion exchange membrane is attached or the A sample concentrating and separating device characterized in that the analyte is concentrated in a region adjacent to a vertical point. According to claim 1,
The size of the micropores in the channel to which the ion exchange membrane is attached is reduced, so that the first analyte having a size smaller than the size of the reduced micropores passes through and the agent having a size larger than the size of the reduced micropores 2 The analyte does not pass, so the lateral flow analysis method characterized in that the analyte is separated. According to claim 1,
The ion exchange membrane is arranged to have a predetermined length along a direction in which the channel extends, and the sample is characterized by controlling the depletion force and depletion region by adjusting the length, thickness, width, or a combination thereof. Concentration and separation device. According to claim 1,
The sample concentration and separation device, characterized in that the position of the concentration of the analyte in the channel is changed according to the presence or absence of a surfactant in the sample. According to claim 1,
A sample concentration and separation device characterized in that the concentration of hydrogen ions (pH) of the analyte moving the channel is adjusted to a predetermined range using a buffer solution having a hydrogen ion concentration set according to the polarity of the ion exchange membrane. According to claim 1,
The ion exchange membrane includes both a cation exchange membrane (CEM) and an anion exchange membrane (AEM),
A device for concentrating and separating a sample, characterized in that, by contacting or spaced apart the cation exchange membrane and the anion exchange membrane, the hydrogen ion concentration (pH) of the analyte moving the channel is adjusted within a predetermined range. According to claim 2,
Sample concentrating and separating device connected to the channel, further comprising a conjugate portion comprising a conjugate in which a detector and an indicator that bind to the analyte form a complex. The method of claim 10,
The detection unit includes a first capturer that captures the complex,
The channel is a sample concentration and separation device, characterized in that it comprises a control comprising a second capturer to capture the complex or the detector failed to form the complex. The method of claim 10,
The ion exchange membrane is a sample concentration and separation device, characterized in that attached over the conjugate portion. According to claim 1,
The ion exchange membrane is a sample concentration and separation device, characterized in that attached to the top, bottom, side, or a combined position of the channel. The method of claim 13,
The number of the ion exchange membranes is plural, and the plurality of ion exchange membranes are arranged in a zigzag manner or formed in a stacked structure, thereby concentrating and separating a sample. According to claim 1,
A device for concentrating and separating a sample, characterized in that a concentrating position control part is located between the ion exchange membrane and the channel. According to claim 1,
The channel may include (i) a straight section, (ii) a curved section, (iii) a'v' shape or a curved section including a'a' shape, (iv) a slope section with different heights, and (v) a'X' shape. Intersecting section comprising, (vi) a branching section containing a'Y' shape, (vii) a stacking section in which a plurality of channels are layered, (viii) a through section in which upper and lower channels are connected via vias, or a combination thereof. Characterized in that the sample concentration and separation device. According to claim 1,
The sample concentrating and separating device further comprises an inlet that is a passage through which the sample flows and an outlet that is a passage through which the sample flows. According to claim 1,
A sample concentrating and separating device further comprising a receiving portion connected to the channel and having a space for receiving the sample. According to claim 1,
A sample concentrating and separating device further comprising an absorber connected to the channel and absorbing the sample by capillary action. According to claim 1,
The sample concentration and separation device is a sample concentration and separation device, characterized in that implemented in a lateral flow array or a microfluidic chip.

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