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WO2016129894A1 - Capteur intégré dans une fonction de concentration de biomolécules et son procédé de fabrication - Google Patents

Capteur intégré dans une fonction de concentration de biomolécules et son procédé de fabrication Download PDF

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Publication number
WO2016129894A1
WO2016129894A1 PCT/KR2016/001306 KR2016001306W WO2016129894A1 WO 2016129894 A1 WO2016129894 A1 WO 2016129894A1 KR 2016001306 W KR2016001306 W KR 2016001306W WO 2016129894 A1 WO2016129894 A1 WO 2016129894A1
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Prior art keywords
layer
electrodes
signal
electrode
impedance
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Korean (ko)
Inventor
황교선
김진식
김태송
이정훈
한성일
유용경
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Korea Institute of Science and Technology KIST
Research Institute for Industry Cooperation of Kwangwoon University
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Korea Institute of Science and Technology KIST
Research Institute for Industry Cooperation of Kwangwoon University
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/333—Ion-selective electrodes or membranes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/301—Reference electrodes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor

Definitions

  • the present specification relates to a biosensor and a method for manufacturing the same, and relates to an electrical microsensor having a biomolecule concentration function and a method for manufacturing the same.
  • Biomarkers are markers that can distinguish between normal and pathological conditions, predict therapeutic response, and measure objectively.
  • biomarkers nucleic acid DNA, RNA (gene), proteins, fats, metabolites, and the like, and changes in their patterns are used.
  • genes such as blood glucose for the diagnosis of diabetes mellitus, BCR-ABL gene fusion of chronic myelogenous leukemia, which is the treatment target of Gleevec, are all biomarkers and are biomarkers that are actually used in clinical practice.
  • DNA (Deoxyribonucleic Acid) is a genetic material that exists in the nucleus, and the gene is where chemical information is stored that determines the type of protein produced by an organism. Information constituting the human body can be grasped by analyzing DNA, and various DNA analysis techniques have been researched and developed and utilized for the prevention and treatment of diseases.
  • PCR Polymerase Chain Reaction
  • PCR repeats heating and cooling with heat-stable DNA polymerase in order to use the single strand formed by successive separation of DNA double helix into a new double helix.
  • the DNA is first divided into two chains by applying heat to DNA. .
  • Proteins are complex molecules made by linking relatively simple molecules called amino acids, which usually have very high molecular weights. There are about 20 types of amino acids that make up proteins, and these amino acids are linked to each other through chemical bonds to make polypeptides. At this time, the binding of amino acids is called a peptide bond, and the peptide is called a polypeptide in the meaning of several (poly-). In a broad sense, a protein is also a polypeptide. Generally, if a molecular weight is relatively small, it is called a polypeptide, and if a molecular weight is very large, a protein is called.
  • Such a protein is a representative molecule constituting the body of the organism, and is a substance responsible for the catalytic role and immunity of various chemical reactions in the cell. Protein is thus a very important organic material that constitutes the living body and participates in the reaction and energy metabolism in vivo.
  • Such DNA or protein may be analyzed to determine the onset and progression of cancer or disease.
  • a blood fingerprint analysis technique is known that finds an indicator protein that shows minute changes in the early stages of development of normal cells into cancer cells among proteins in blood.
  • the blood fingerprint analysis is based on the idea that metabolites in the human body can be changed depending on the presence or absence of cancer, and comprehensively analyzes mass spectrometry data of metabolites in the blood of cancer patients to diagnose cancer occurrence through trends in patterns. It is a technique.
  • the technologies and devices for protein analysis currently introduced have a problem that it is difficult to manufacture the device by using nanotechnology and it is difficult to be widely used.
  • the protein analysis device requires a high-sensitivity sensor or has a disadvantage in that accurate analysis is difficult with a small sample.
  • the present invention has been proposed to solve the above problems, and an object of the present invention is to provide a biosensor with a biomolecule enrichment function and a method of manufacturing the same.
  • Another object of the present invention is to provide a biosensor capable of easily detecting an extremely low concentration of a target substance.
  • the biomolecule enrichment function integrated sensor includes a microchannel, a reservoir for receiving a sample of a target material formed on both ends of the microchannel, and an optional ion permeable membrane pattern.
  • Thickener comprising a; And a cross-electrode portion formed on the substrate so that the two electrodes form a slot and are spaced apart from each other by a predetermined interval, wherein the target material concentrated in a predetermined region on the microchannel is formed in the slot formed by the two electrodes.
  • the detector is characterized in that the receptor for capturing the target material is installed in the slot formed by the two electrodes.
  • the concentrator includes the first ion-permeable membrane pattern, the first layer in which the reservoir is formed, and the second layer in which the microchannel is formed, and the selective ion-permeable membrane pattern formed in the first layer is the second layer. And mutually intersecting the microchannel formed in the layer.
  • a positive electrode and a negative electrode may be connected to both ends of the reservoir, respectively, so that a potential difference may be generated at both ends of the microchannel.
  • the specific difference of the target material sample of the reservoir passes through the microchannel by the potential difference.
  • the cross-electrode portion of the detector Concentrated in a predetermined region of the front end of the selective ion permeable membrane, the cross-electrode portion of the detector is characterized in that the pattern is formed corresponding to the predetermined region of the micro channel.
  • the thickener and the sensor is characterized in that coupled through a transfer film (transfer film).
  • the detector may further include: a driving signal applying unit configured to apply a driving frequency signal having a predetermined frequency to one of the two electrodes; And an impedance measuring unit measuring an impedance by using a signal output to the other electrode of the two electrodes.
  • the detector a channel through which current flows between the two electrodes is formed, a protective cap for protecting the receptor installed for capture of the target material is installed in the slot formed by the two electrodes, the receptor It is characterized in that the adsorption prevention layer is coated on the inner wall of the protective cap and the surface of the two electrodes is not installed.
  • the cross-electrode portion of the detector may include a reference cross-electrode portion in which the target material is not captured, and a signal cross-electrode portion in which the target material is captured by a receptor provided between the two electrodes.
  • the detector may include: a driving signal applying unit configured to apply a driving frequency signal having a predetermined frequency to one of two electrodes of each of the reference cross electrode unit and the signal cross electrode unit; A reference impedance measuring unit measuring an impedance between two electrodes of the reference cross electrode unit as a reference impedance from a signal sensed by the other electrode of the two electrodes of the reference cross electrode unit; A signal impedance measuring unit measuring an impedance between two electrodes of the signal cross electrode unit as a signal impedance from a signal detected by the other electrode of the two electrodes of the signal cross electrode unit; And a differential amplifier configured to differentially amplify the reference impedance and the signal impedance from the reference impedance measuring unit and the signal impedance measuring unit, respectively.
  • the selective ion permeable membrane may be formed of one of nafion, polystyrene sulfonate (PSS) or polyallylamine hydrochloride (PAH).
  • PSS polystyrene sulfonate
  • PAH polyallylamine hydrochloride
  • the biomolecule enrichment function integrated sensor includes a first layer in which a reservoir for receiving a selective ion permeable membrane pattern and a target material sample is formed; A second layer having both ends connected to the reservoir to form a micro channel, the second layer having an upper end coupled to the bottom of the first layer; And two electrodes coupled to a lower end of the second layer and formed on the substrate so as to be spaced apart from each other at predetermined intervals to form slots therebetween, the cross electrodes being disposed at positions corresponding to the predetermined region of the microchannel. And a third layer having a portion.
  • a method for manufacturing a biomolecule enrichment function integrated sensor forms a selective ion permeable membrane using a microflow patterning technique on an upper surface of a substrate. step; Forming a PDMS mold on the upper surface of the substrate by pouring polydimethylsiloxane (PDMS) on the upper surface of the substrate on which the selective ion permeable membrane is formed; Separating the substrate and the PDMS mold to form a first PDMS layer including the selective ion permeable layer in the separated PDMS mold; Forming a reservoir in the first PDMS layer; Bonding a second PDMS layer having a microchannel to the first PDMS layer to which the selective ion permeable membrane is exposed; And forming an electrode pattern on the substrate such that the two electrodes form a slot and are spaced apart from each other to form a cross electrode portion, and joining the substrate on which the cross electrode portion is generated to the second PDMS layer.
  • PDMS polydimethylsiloxane
  • the selective ion-permeable membrane included in the first PDMS layer and the microchannel included in the second PDMS layer may mutually intersect at an interface between the first PDMS layer and the second PDMS layer.
  • the concentrated function integrated sensor according to an embodiment of the present specification, by forming an ion permeable membrane using a material capable of selective ion permeation, there is an excellent effect of protein concentration efficiency even without using a nanochannel.
  • the concentrated function integrated sensor according to an embodiment of the present specification has the effect of accurately detecting the presence and concentration of a biological material through impedance measurement without using conductive particles.
  • FIG. 1 is a perspective view of a biomolecule enrichment function integrated sensor according to an embodiment of the present specification.
  • Figure 2 is an exploded perspective view of a biomolecule enrichment function integrated sensor according to an embodiment of the present disclosure.
  • FIG 3 is a view showing the electrode pattern of the sensor in the biomolecule enrichment function integrated sensor according to an embodiment of the present disclosure.
  • FIG. 4 is a cross-sectional view of a detector in a biomolecule enrichment integrated sensor according to an embodiment of the present disclosure.
  • FIG. 5 is a block diagram of a detector to which a differential amplifier is applied according to an embodiment of the present specification.
  • FIG. 6 is a reference view for explaining a method of manufacturing a sensor in a biomolecule enrichment function integrated sensor according to one embodiment of the present specification.
  • FIG. 7 is a reference diagram for explaining a method of manufacturing a first layer of a concentrator in a biomolecule concentrating function integrated sensor according to one embodiment of the present specification.
  • FIG. 8 is a reference diagram for explaining a method of manufacturing a second layer of a concentrator in a biomolecule concentrating function integrated sensor according to one embodiment of the present specification.
  • FIG. 9 is a reference diagram for explaining a process of combining a concentrator and a sensor in a biomolecule concentrating function integrated sensor according to one embodiment of the present specification.
  • FIG. 1 is a perspective view of a biomolecule enrichment function integrated sensor 500 according to an embodiment of the present specification.
  • the senor 500 is a concentrator 100 having a selective ion permeable membrane pattern 12 and a micro channel 52 and a target material through the concentrator 100. It includes a detector 200 that detects the concentration when concentrated in the area.
  • the concentrator 100 includes a microchannel 52, a target material sample reservoir 16 formed across the microchannel, and an optional ion permeable membrane pattern 12.
  • both inlets of the microchannel 52 are formed with reservoirs 16 for receiving a protein sample to be analyzed, respectively, and the reservoirs 16 are wires (not shown) that can be connected to an external power source, respectively. ) Is formed.
  • the negative electrode and the positive electrode are respectively connected to the reservoir 16 through the wire, so that a potential difference is generated at both ends based on the selective ion permeable membrane 12.
  • the reservoir 16 may be provided with a thin film electrode to be connected to an external power source.
  • the selective ion-permeable membrane 12 patterned on the PDMS layer 10 outside of the microchannel 52 and intersecting with the microchannel serves as a kind of nanofilter that selects and transmits protons. .
  • the detector 200 includes a cross-electrode portion 20 formed on a substrate so that two electrodes form a slot and are spaced apart from each other by a predetermined distance, and the target is concentrated in a predetermined region on the micro channel 52.
  • the material can be captured by a receptor (not shown) disposed in the slot formed by the two electrodes.
  • FIG 2 is an exploded perspective view of a sensor 500 according to an embodiment of the present disclosure.
  • the senor 500 has a form in which the concentrator 100 and the sensor 200 are combined, and the three layers structure is combined.
  • the concentrator 100 includes the first layer 10 having the reservoir 16 and the selective ion permeable membrane pattern 12 therein, and the second layer 50 having the microchannel 52 formed therein.
  • the detector 200 is a third layer 200 that is coupled to the bottom of the second layer 50, and the cross-electrode portion 20 having a pattern formed on the substrate so that the two electrodes form a slot and are spaced apart from each other by a predetermined interval is provided. Is formed.
  • the selective ion permeable membrane 12 patterned inside the first layer 10 and intersecting with the microchannel 52 of the second layer 50 serves as a kind of nano filter that selects and transmits protons. do.
  • H + ions are caused by the H3- ions in the chemical structure of Nafion and the hopping and vehicle mechanism (K.-. D. Kreuer, Chem. Mater. 1996, 8, 610-641).
  • the selective ion permeable membrane 12 provides a function of causing the microchannel 52 to function substantially as a nanochannel, and the protein materials to be analyzed at the front end toward the positive electrode of the nanochannel are very fast. It can be concentrated efficiently.
  • the device is configured such that the PDMS layer 10 in which the selective ion permeable material is patterned is disposed on the upper surface of the micro-sized channel 52. I can make it.
  • the microchannel 52 is externally formed, the microchannel 52 performs the function of the nanochannel through the selective ion permeable membrane 12.
  • EDL electric double layer
  • the characteristics of capillary electrophoresis and electro-osmosis are different near the nanochannel and ion concentration polarization near the nanochannel or the selective ion permeable membrane 12.
  • ion enrichment occurs at the cathode side and ion depletion at the anode side based on the nanochannel.
  • the depletion zone acts as a kind of electric barrier to the charged protein due to the low ion concentration and the high electric field.
  • the protein does not pass through the deficient region and is concentrated before it.
  • the third layer coupled to the bottom of the second layer 50 in which the micro channel 52 is formed performs the function of the detector 200 for detecting the concentration of the target material.
  • the sensor 200 forms a predetermined electrode pattern on a substrate, and a receptor for capturing a target material is disposed between the electrodes to react with a chemical or biological target material, and detects a change in the receptor to detect an electrical signal. It can be configured to include a transducer (transducer) to convert to.
  • FIG 3 is a view showing the electrode pattern 20 of the sensor 200 according to an embodiment of the present disclosure
  • Figure 4 is a biomolecule enrichment function integrated sensor according to an embodiment of the present specification of the sensor 200 It is a figure which shows a cross section.
  • the cross-electrode portion 20 is provided on the substrate 60.
  • the interdigitated microelectrode part (IME part) 20 is installed in a form in which two electrodes 21 and 22 having a comb shape are interlocked with each other at a predetermined interval.
  • Equation 1 Z is impedance, R is resistance, X is reactance, C is capacitance, and w is angular frequency.
  • the reactance X is divided into the inductor component XL and the capacitor component XC. Since the two electrodes 21 and 22 are not electrically connected directly, the inductor component XL can be ignored so that only the capacitor component XC exists. Can be.
  • a receptor (mainly an antibody, an aptamer, etc.) 31 which specifically reacts with a target biomaterial 32 is fixed in a space between two electrodes 21 and 22 and the target biomaterial Confirmation of the impedance change between the two electrodes 21 and 22 when the reference numeral 32 responds to the receptor 31 enables the quantitative analysis of the target biomaterial 32.
  • the water (or buffer solution, serum, blood, etc.) existing between the two electrodes 21 and 22 becomes specific.
  • the resistance is increased because the target biomaterial 32 is positioned to push out.
  • the reactance decreases the value of the capacitance C due to the property of the target biological material 32 whose dielectric constant is smaller than that of water (or buffer solution, serum, blood, etc.), thereby increasing the value of the inductor component XC.
  • the -XC value will decrease.
  • the amount of change in the impedance can be checked to accurately detect the amount of the target biomaterial 32.
  • the impedance between the electrodes 21 and 22 of the cross-electrode portion 20 in the detector 200 applies a driving frequency signal having a predetermined driving frequency to one of the two electrodes 21 and 22.
  • the signal output from the remaining electrode 22 to which the driving frequency signal is not applied may be analyzed and measured.
  • the frequency of the driving frequency signal applied to the detector 200 should be high to make it easy to check the change in impedance, and the frequency is low.
  • the change in the detected signal is insignificant, making it difficult to confirm the change. Therefore, in order to detect a small amount of the target biomaterial 32, a high frequency driving frequency signal may be used.
  • the frequency of the driving frequency signal when the frequency of the driving frequency signal is high, current mainly flows through the upper space of the specifically coupled target biomaterial 32, that is, the A channel A shown in FIG. Detection will not work properly. In addition, if the frequency is high, the target biomaterial 32 may be damaged by high frequency and may not be detected properly.
  • the current flows through the B channel B.
  • driving of approximately 10 Hz to 100 Hz is performed so that the current can flow through the channel B. It is characterized by using a frequency signal.
  • a frequency signal When using a low frequency drive frequency signal in this way, because the frequency is low it can be prevented that the target biomaterial 32 is damaged.
  • the frequency of the driving frequency signal is low, there is a disadvantage in that it is difficult to detect a fine impedance change in the B channel (B), but this disadvantage can be overcome by using a differential amplifier as described below.
  • the gap between the two electrodes 21 and 22 is approximately 3-7 ⁇ m. Is preferably. This is because, if the gap is smaller than 3 mu m, the detection signal is too large, so that a reliable test cannot be performed. If the gap is too large, it is insufficient to detect a small amount of the biomaterial 32 because the gap is larger than 7 mu m. . Considering the deviation and sensitivity, the case of 5 ⁇ m is most preferable.
  • FIG. 5 is a block diagram of a sensor to which a differential amplifier is applied according to an embodiment of the present specification.
  • the signal cross electrode unit 120 and the reference cross electrode unit 220 are installed on the substrate 60, and each of the signal cross electrode unit 120 and the reference cross electrode unit 220 is illustrated in FIG. 3.
  • Two electrodes 21 and 22 having a comb-like shape, such as the cross-electrode portion 20, are spaced apart from each other by a predetermined interval and are arranged in a staggered manner.
  • the capture of the target biomaterial 32 is performed by the receptor 31 installed between the two electrodes 21 and 22, while the capture in the reference cross-electrode unit 220 is not performed. Do not. That is, the signal cross-electrode unit 120 allows the electrode pattern to be formed in a region where a specific target material is concentrated in a predetermined region in front of the selective ion permeable membrane 12 of the microchannel 52 of the concentrator 100, and the reference The cross-electrode portion 220 is formed with a pattern to be located in a region where the target material of the micro channel 52 is not concentrated.
  • the target biomaterial 32 may be provided only to the signal cross-electrode unit 120.
  • the receptor 31 is installed only on the signal cross electrode unit 120, and the receptor 31 is not installed on the reference cross electrode unit 220, and both the signal cross electrode unit 120 and the reference cross electrode unit 220 are provided.
  • the target biomaterial 32 may be provided to the target biomaterial 32 so that the capture of the target material is possible only at the signal cross-electrode unit 120.
  • a driving frequency is applied to one of the two electrodes 21 and 22 of the signal cross electrode unit 120 and the reference cross electrode unit 220 through the driving signal applying unit 1.
  • the signal impedance measuring unit 125 receives a signal applied through the remaining electrodes 22 of the signal cross electrode unit 120 and measures the impedance of the signal cross electrode unit 120 as the signal impedance.
  • the reference impedance measuring unit 225 receives a signal applied through the remaining electrodes 22 of the reference cross-electrode unit 220 and measures the impedance of the reference cross-electrode unit 220 as a reference impedance.
  • the signal impedance will be the impedance between the two electrodes 21 and 22 for the case where the target biomaterial 32 is captured between the two electrodes 21 and 22, and the reference impedance will be between the two electrodes 21 and 22. Will be the impedance between the two electrodes 21, 22 for the case where the target biomaterial 32 is not captured.
  • the differential amplifier 300 receives the signal impedance and the reference impedance, respectively, and differentially amplifies the signal impedance and outputs a result signal.
  • the impedance change in the B channel B is insignificant with respect to the driving frequency signal in the low frequency range of 10 Hz to 100 Hz. (32) is difficult to detect.
  • the signal cross-electrode part 120 and the reference cross-electrode part 220 are provided to differentially amplify the impedance of the signal cross-electrode part 120 based on the impedance of the reference cross-electrode part 220.
  • FIG. 6 is a reference diagram for describing a manufacturing process of the sensor 200 according to an exemplary embodiment of the present specification.
  • a 500 nm thick silicon oxide film SiO 2, 61 is formed on the silicon substrate 60 by thermal oxidation, and then sputtered on the silicon oxide film 61.
  • a metal layer 20a is formed by sequentially stacking 30 nm of titanium (Ti) and 150 nm of platinum (Pt) by a sputtering method.
  • the titanium (Ti) layer is used as an adhesion layer for increasing the bonding force between the platinum (Pt) layer and the silicon oxide film 11.
  • a photoresist film is coated on the metal layer 20a and the photoresist film is patterned by a photolithography process to form a photoresist pattern 40.
  • the metal layer 20a is formed using an inductively coupled plasma reactive ion etcher (ICP-RIE) until the silicon oxide layer 61 is exposed using the photoresist pattern 40 as an etch mask. 2) to form the two electrodes 21 and 22, and then, the photoresist pattern 40 is removed as shown in FIG.
  • ICP-RIE inductively coupled plasma reactive ion etcher
  • Specific binding of the target biomaterial 32 using the electrode pattern thus formed is performed as follows.
  • a Calixcrown SAM Self-Assembled Monolayer as a linking molecule layer 33 for selectively fixing beta amyloid antibody on the surface of the silicon oxide film 61 between the two electrodes 21 and 22.
  • the beta amyloid antibody is immobilized on the linker molecule layer 33 as the receptor 31.
  • the beta amyloid that is the target biomaterial 32 is selectively specifically bound to the receptor 31.
  • a protective cap (not shown) is preferably installed on the two electrodes 21 and 22 so that the two electrodes 21 and 22 are placed in the channel.
  • the channel by the protective cap also helps the sample enter the specific binding region. It is preferable to use a protective cap of PDMS (Polydimethylsiloxane) material.
  • the target biomaterial 32 When a sample containing various components is introduced into the channel, only the target biomaterial 32 that specifically reacts with the receptor 31 is bound to the receptor 31.
  • the target biomaterial 32 will be beta amyloid.
  • an adsorption prevention layer (not shown) is coated on the surface.
  • the adsorption prevention layer here is preferably made of BSA (Bovine Serum Albumin).
  • the detector 200 By configuring the detector 200 as described above, it is possible to accurately detect the presence and concentration of the biological material through the impedance measurement without using the conductive particles as in the prior art. In addition, there is an advantage that the precise detection can be made while using a low frequency of 10Hz ⁇ 100Hz so that the target biomaterial 32 is not denatured or damaged. In addition, by using the differential amplifier 300, there is an advantage that precision detection is possible even for a small amount of biomaterial.
  • FIG. 7 is a reference diagram for explaining a process of manufacturing a first layer of a concentrator according to one embodiment of the present specification.
  • a substrate 11 having a rectangular shape having a predetermined thickness and area is formed.
  • the substrate 11 may be formed of a glass substrate, a plastic substrate such as polycarbonate (PC) or polydimethylsiloxane (PDMS), or a silicon substrate.
  • the upper portion of the substrate is preferably surface-treated using Teflon or the like on the surface.
  • a permselective membrane 12 capable of selective ion permeation is formed on the top surface of the Teflon-treated substrate 11 by using a microflow patterning method.
  • the permselective membrane 12 is a membrane that selectively permeates specific ions, hereinafter referred to as selective ion permeable membrane 12.
  • the selective ion permeable membrane 12 is formed of a material capable of selective ion permeation, and typically Nafion is used.
  • polyelectrolyte such as polystyrene sulfonate (PSS) and polyallylamine hydrochloride (PAH) may be applied. It's okay.
  • the selective ion-permeable membrane 12 patterned on the upper surface of the substrate 11 is formed of a Nafion membrane 12 having a predetermined thickness and length through a microflow patterning technique and is hundreds of times. It can be formed to have a thickness of several tens of micrometers in the nano.
  • the selective ion-permeable membrane 12 may be formed on the top surface of the substrate 11 by using a bonding technique without using the same microflow patterning technique, but in this case, leakage due to incomplete bonding may occur. Can be generated.
  • the selective ion permeable membrane 12 may have a width of approximately 500 ⁇ m and a height of 50 ⁇ m.
  • an oxide layer may be formed by depositing an oxide such as SiO 2 on the surface of the selective ion permeable membrane 12.
  • an oxide such as SiO 2
  • a metal may be deposited on the surface of the oxide.
  • the metal may be deposited on the surface of the oxide using Au, Pt and the like.
  • the deposited metal film or metal layer serves as an electrode, and controls the amount of ions passing through the Nafion membrane or the surface-potential and zeta-potential of the Nafion membrane. Therefore, the selective ion permeable film 12, the oxide layer (not shown), and the metal layer (not shown) may be patterned on the upper surface of the substrate 11 in order.
  • an electrode pattern is formed on both surfaces of the PDMS layer 10 so that a part of the metal thin film layer is exposed to the outside, and thus charge may be supplied or exhausted to the metal layer. For example, as the charge is exhausted through the electrode pattern, negative charge is formed inside the metal layer.
  • positive charges are formed in the oxide layer in contact with the metal layer, and negative charges are formed in the oxide layer in contact with the surface of the selective ion permeable membrane 12 on the opposite side. In order to compensate for the negative charge on the oxide surface to maintain electrical neutrality, the selective ion permeable membrane retains more cations in the fluid.
  • the ions passing through the selective ion permeable membrane You can adjust the amount.
  • the amount of ions passing through the selective ion permeation membrane is directly related to the intensity of the ion concentration polarization phenomenon, and by controlling this, the degree of concentration can be controlled.
  • the zeta potential here refers to an electrodynamic potential difference resulting from the difference in positive charge density in the diffusion double layer of immobilized moisture on the charged particle surface and movable moisture that can be easily separated from the particle.
  • the colloidal particles move (move) in the direction opposite to the sign of the surface potential.
  • the electric field strength and the hydrodynamic effect (solvent viscosity, dielectric constant, etc.) applied the particle movement speed at this time Calculated in consideration of the zeta potential.
  • the PDMS mold 10 is formed by pouring PDMS (polydimethylsiloxane) on the upper surface of the substrate 11 on which the selective ion permeable membrane 12 is patterned.
  • the mold 10 is formed by using the PDMS.
  • the selective ion permeable layer 12 is patterned by pouring a polymer such as PD or PC in addition to the PDMS. It is also possible to form a mold 10 on the top surface.
  • a PDMS layer 10 including the selective ion permeable layer 12 is formed in the PDMS.
  • a surface treatment using Teflon or the like is performed on the upper surface of the substrate 11 as described above. It is desirable to.
  • a reservoir 16 is formed in the PDMS layer 10 to receive a target material (eg, protein) sample to be analyzed, and each of the wires 16 may be connected to an external power source. (Not shown) is formed.
  • the negative electrode and the positive electrode are respectively connected to the reservoir through the wire, so that a potential difference is generated at both ends based on the selective ion permeable membrane 12.
  • the reservoir may be provided with a thin film electrode to be connected to an external power source.
  • a second layer 50 having a microchannel connected to the reservoir 16 while contacting the selective ion permeable membrane 12 is formed to be bonded with the first layer.
  • FIG 8 is a reference diagram for explaining a second layer manufacturing process of the concentrator 100 according to one embodiment of the present specification.
  • a thin film PDMS 53 is deposited on an upper surface of a substrate 51 having a predetermined thickness and area.
  • a transfer film (transfer film) 55 is attached to the PDMS 53.
  • the PDMS layer 53 to which the transfer film 55 is attached is separated from the substrate 51.
  • a micro channel 52 is formed inside the PDMS layer. Both ends of the micro channel 52 are connected to each of the reservoirs 16 formed in the first layer, and the selective ion permeable membrane 12 and the micro channel 52 formed in the first layer are in contact with each other. do.
  • the microchannel 52 may have a width of about 100 ⁇ m and a thickness of about 50 ⁇ m.
  • FIG. 9 is a reference diagram for explaining a coupling process of the concentrator 100 and the sensor 200 according to an exemplary embodiment of the present specification.
  • the first layer 10 described above with reference to FIG. 7, the second layer 50 described with reference to FIG. 8, and the third layer (ie, the detector) 200 described with reference to FIG. 6 are sequentially layered. Bonding and coupling form the sensor 500 herein.
  • the surface on which the selective ion permeable membrane 12 of the first layer 10 is formed and the PDMS layer of the second layer 50 are bonded to each other.
  • the layer 10 and the second layer 50 are combined.
  • the transfer film 55 of the second layer 50 is coupled to face the lower surface to be combined with the third layer.
  • the concentrator and the detector are combined by applying heat or energy, the receptor may be damaged. In this embodiment, the concentrator and the detector are combined without applying heat or energy.
  • the sensor 500 may further include a driving signal applying unit for applying a driving frequency signal between two electrodes of the third layer and an impedance measuring unit for measuring impedance between the two electrodes of the third layer.

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Abstract

La présente invention se rapporte à un capteur intégré dans une fonction de concentration de biomolécules. Selon un mode de réalisation de la présente invention, le capteur comprend : un concentrateur comprenant un microcanal, un réservoir formé au niveau des deux extrémités du microcanal et recevant un échantillon de matériau cible, et un motif de membrane perméable sélective aux ions; et un capteur comprenant une partie de microélectrode interdigitée (IME pour Interdigitated MicroElectrode), qui présente un motif formé sur un substrat de telle sorte que deux électrodes soient séparées l'une de l'autre selon un intervalle prédéterminé tout en formant une fente entre elles, et permettant la capture du matériau cible, qui est concentré dans une région prédéterminée sur le microcanal, au niveau de la fente formée par les deux électrodes.
PCT/KR2016/001306 2015-02-09 2016-02-05 Capteur intégré dans une fonction de concentration de biomolécules et son procédé de fabrication Ceased WO2016129894A1 (fr)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018057647A1 (fr) * 2016-09-23 2018-03-29 Alveo Technologies, Inc. Méthodes et compositions de détection d'anévrismes
EP3399305A4 (fr) * 2015-12-28 2019-08-28 Korea Institute of Science and Technology Bio-capteur à électrode inter-digitée utilisant une réaction entre un récepteur et un biomatériau cible
EP3950132A1 (fr) * 2020-08-04 2022-02-09 Technische Universität Wien Procédé de détection et de quantification d'analytes dans un dispositif microfluidique
EP3818361A4 (fr) * 2018-07-03 2022-04-06 Curi Bio, Inc. Dispositifs et procédés de migration cellulaire à guidage topographique
CN114415078A (zh) * 2021-12-07 2022-04-29 中国科学院上海微系统与信息技术研究所 微型仿生弱磁场传感器及其制备方法和磁场测试方法
US11473128B2 (en) 2014-10-06 2022-10-18 Alveo Technologies, Inc. System and method for detection of nucleic acids
US12275007B2 (en) 2018-12-20 2025-04-15 Alveo Technologies, Inc. Handheld impedance-based diagnostic test system for detecting analytes
US12472492B2 (en) 2020-08-14 2025-11-18 Alveo Technologies, Inc. Systems and methods of sample depositing and testing

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11506580B2 (en) 2016-09-28 2022-11-22 Calth. Inc. Sample separation device based on paper folding
KR101968319B1 (ko) * 2017-08-22 2019-04-11 광운대학교 산학협력단 여과층을 이용한 종이접기 기반의 시료 분리 장치
KR101912890B1 (ko) * 2016-10-04 2018-10-29 한국과학기술연구원 표적 생체물질과 수용체의 반응을 개선한 교차 전극 바이오센서
KR101924415B1 (ko) * 2017-05-02 2019-02-20 한국과학기술원 복합 임피던스 측정 장치 및 측정 방법
KR102102534B1 (ko) * 2018-07-11 2020-04-23 주식회사 엑스와이지플랫폼 유전 전기 영동을 이용한 마이크로 전극 바이오 센서, 및 이를 이용한 생체물질 검출 방법
KR102181866B1 (ko) * 2019-07-23 2020-11-23 한국광기술원 나노물질 네트워크 소자를 이용한 미세먼지 감지 장치 및 방법
KR102350449B1 (ko) * 2019-11-15 2022-01-11 광운대학교 산학협력단 정전용량을 기반한 바이오센서 및 그의 제조방법
RU2745663C1 (ru) * 2019-12-31 2021-03-30 федеральное государственное бюджетное образовательное учреждение высшего образования "Московский государственный технический университет имени Н.Э. Баумана (национальный исследовательский университет)" (МГТУ им. Н.Э. Баумана) Способ изготовления матричного биосенсора на основе восстановленного оксида графена и матричный биосенсор на полимерной подложке

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090101764A (ko) * 2008-03-24 2009-09-29 디지탈 지노믹스(주) 생리활성물질을 전기적으로 검출하는 방법 및 이를 위한바이오칩
KR20100045640A (ko) * 2008-10-24 2010-05-04 재단법인대구경북과학기술원 교환 가능한 전극을 구비하는 ｆｅｔ 형 바이오 센서
KR20100087130A (ko) * 2007-09-26 2010-08-03 메사추세츠 인스티튜트 오브 테크놀로지 전기역학적 농축 장치 및 그의 사용 방법
KR101248271B1 (ko) * 2012-02-21 2013-03-27 광운대학교 산학협력단 마이크로-나노 채널을 이용한 에너지 변환 소자 및 그 제조 방법
KR20130095920A (ko) * 2012-02-21 2013-08-29 광운대학교 산학협력단 마이크로-나노 채널을 이용한 3차원 에너지 변환 소자 및 그 제조 방법

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20100087130A (ko) * 2007-09-26 2010-08-03 메사추세츠 인스티튜트 오브 테크놀로지 전기역학적 농축 장치 및 그의 사용 방법
KR20090101764A (ko) * 2008-03-24 2009-09-29 디지탈 지노믹스(주) 생리활성물질을 전기적으로 검출하는 방법 및 이를 위한바이오칩
KR20100045640A (ko) * 2008-10-24 2010-05-04 재단법인대구경북과학기술원 교환 가능한 전극을 구비하는 ｆｅｔ 형 바이오 센서
KR101248271B1 (ko) * 2012-02-21 2013-03-27 광운대학교 산학협력단 마이크로-나노 채널을 이용한 에너지 변환 소자 및 그 제조 방법
KR20130095920A (ko) * 2012-02-21 2013-08-29 광운대학교 산학협력단 마이크로-나노 채널을 이용한 3차원 에너지 변환 소자 및 그 제조 방법

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11473128B2 (en) 2014-10-06 2022-10-18 Alveo Technologies, Inc. System and method for detection of nucleic acids
EP3399305A4 (fr) * 2015-12-28 2019-08-28 Korea Institute of Science and Technology Bio-capteur à électrode inter-digitée utilisant une réaction entre un récepteur et un biomatériau cible
WO2018057647A1 (fr) * 2016-09-23 2018-03-29 Alveo Technologies, Inc. Méthodes et compositions de détection d'anévrismes
US11465141B2 (en) 2016-09-23 2022-10-11 Alveo Technologies, Inc. Methods and compositions for detecting analytes
EP3818361A4 (fr) * 2018-07-03 2022-04-06 Curi Bio, Inc. Dispositifs et procédés de migration cellulaire à guidage topographique
US12275007B2 (en) 2018-12-20 2025-04-15 Alveo Technologies, Inc. Handheld impedance-based diagnostic test system for detecting analytes
EP3950132A1 (fr) * 2020-08-04 2022-02-09 Technische Universität Wien Procédé de détection et de quantification d'analytes dans un dispositif microfluidique
WO2022029160A3 (fr) * 2020-08-04 2022-07-07 Technische Universität Wien Procédé de détection et de quantification d'analytes dans un dispositif microfluidique
US12472492B2 (en) 2020-08-14 2025-11-18 Alveo Technologies, Inc. Systems and methods of sample depositing and testing
CN114415078A (zh) * 2021-12-07 2022-04-29 中国科学院上海微系统与信息技术研究所 微型仿生弱磁场传感器及其制备方法和磁场测试方法

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