Dual Immunodiagnostics Chip

Abstract

The present invention provides a dual immunodiagnostic chip. By using the dual immunodiagnostic chip, changes in color and electrical signals can be observed through a single sample input. Through this, the presence or absence of a target substance can be detected rapidly and accurately. The dual immunodiagnostic chip of the present invention comprises: an injection section (100) into which a composite nanostructure, comprising a sample, a nano particle, and a first bioreceptor, is injected; a reaction section (200) coated with a color-changing reagent that reacts with the composite nanostructure to emit color; an electrode section (300) in which a second bioreceptor capable of binding to a target substance is fixed and generates an electrical signal; and a microfluidic channel section (400) connected to the injection section (100), the reaction section (200), and the electrode section (300), and providing a passage so that the composite nanostructure injected into the injection section (100) can flow into the reaction section (200) and the electrode section (300).

Description

Technology Field

[0001] The technology described below relates to a dual immune diagnostic chip. Background Technology

[0002] Microfluidics-based technology involves the manipulation of fluids in small volumes. Microfluidics is utilized in various fields, including biotechnology, medicine, environmental science, and physics. Recently, technologies utilizing microfluidics to detect the presence of target substances within samples are being developed. In particular, immunodiagnostic chips capable of detecting cancer or viruses are being utilized by combining composite nanostructure technology, in which antibodies are bound to functional nanoparticles. Immunodiagnostic chips can detect cancer or viruses by operating in such a way that changes in optical or electrochemical signals vary depending on the presence or absence of target substances. Prior art literature

[0003] Korean Registered Patent 10-1188172 The problem to be solved

[0004] One aspect is to provide a dual immune diagnostic chip.

[0005] Another aspect is to provide a method for detecting target substances using a dual immunodiagnostic chip. means of solving the problem

[0006] The dual immunodiagnostic chip comprises: an injection section (100) into which a composite nanostructure, comprising a sample, a nano particle, and a first bioreceptor, is injected; a reaction section (200) coated with a color-changing reagent that reacts with the composite nanostructure to emit color; an electrode section (300) in which a second bioreceptor capable of binding to a target substance is fixed and which generates an electrical signal; and a microfluidic channel section (400) connected to the injection section (100), the reaction section (200), and the electrode section (300), which provides a passage so that the composite nanostructure injected into the injection section (100) can flow into the reaction section (200) and the electrode section (300).

[0007] A method for detecting a target substance using a dual immunodiagnostic chip comprises: a step of injecting a composite nanostructure, in which a sample, a nanoparticle, and a first bioreceptor are combined, into an injection part (100); and a step of detecting whether a target substance is present in the sample based on the result of measuring (a) or (b) below. (a) a color change of the reaction part (200) of the dual immunodiagnostic chip; (b) a change in the electrical signal of the electrode part (300) of the dual immunodiagnostic chip. Effects of the invention

[0008] Using the technique described below, it is possible to detect whether a target substance is present in a sample.

[0009] By utilizing the technology described below, the presence of a target substance within a sample can be detected by observing changes in color and electrical signals with a single sample input. This enables rapid and accurate detection of the target substance even when using a small amount of sample. Brief explanation of the drawing

[0010] Figure 1 is an overall overview of the dual immunodiagnostic chip. FIG. 2 is a perspective view of one embodiment of a dual immune diagnostic chip (10). FIG. 3 is an exploded view of one embodiment of a dual immune diagnostic chip (10). FIG. 4 is an example of the first layer of a dual immune diagnostic chip (10). FIG. 5 is an example of the second layer of a dual immune diagnostic chip (10). FIG. 6 is an example of the third layer of a dual immune diagnostic chip (10). FIG. 7 is an example of an electrode part (300). FIG. 8 is an example of an electrode part (300). FIG. 9 is a plan view of one embodiment of a dual immune diagnostic chip (10). FIG. 10 is an example of a reaction that may occur in the reaction section (200). FIG. 10 is an enlarged view of a part of FIG. 1. FIG. 11 is an example of a reaction that may occur in the electrode portion (300). FIG. 11 is an enlarged view of a portion of FIG. 1. FIG. 12 is an example of a change in electrical signal that may occur in the electrode portion (300). Specific details for implementing the invention

[0011] Embodiments are described in detail below with reference to exemplary drawings. It should be noted that when assigning reference numerals to the components of each drawing, the same components are assigned the same reference numeral whenever possible, even if they are shown in different drawings. Furthermore, in describing the embodiments, if it is determined that a detailed description of related known configurations or functions would hinder understanding of the embodiments, such detailed description is omitted.

[0013] The inventors have developed a dual immunodiagnostic chip (Fig. 1). The inventors have developed a method for detecting target substances using the dual immunodiagnostic chip. The dual immunodiagnostic chip allows for simultaneous optical and electrochemical analysis with a single sample input. This enables rapid and accurate detection of target substances even with only a small amount of sample.

[0014] The present invention will be described in detail below.

[0016] One aspect of the present invention provides a dual immune diagnostic chip (10).

[0017] FIG. 2 is a perspective view of one embodiment of a dual immune diagnostic chip (10). FIG. 3 is an exploded view of one embodiment of a dual immune diagnostic chip (10). FIG. 4 is one embodiment of a first layer of the dual immune diagnostic chip (10). FIG. 5 is one embodiment of a second layer of the dual immune diagnostic chip (10). FIG. 6 is one embodiment of a third layer of the dual immune diagnostic chip (10). FIG. 7 is one embodiment of an electrode section (300). FIG. 8 is one embodiment of an electrode section (300). FIG. 9 is a plan view of one embodiment of a dual immune diagnostic chip (10). FIG. 10 is an example of a reaction that may occur in a reaction section (200). FIG. 11 is an example of a reaction that may occur in an electrode section (300). FIG. 12 is an example of a change in an electrical signal that may occur in an electrode section (300).

[0018] A dual immunodiagnostic chip (10) comprises: an injection section (100) into which a composite nanostructure, comprising a sample, a nano particle, and a first bioreceptor, is injected; a reaction section (200) coated with a color-changing reagent that reacts with the composite nanostructure to produce color; an electrode section (300) in which a second bioreceptor capable of binding to a target substance is fixed and generates an electrical signal; and a microfluidic channel section (400) connected to the injection section (100), the reaction section (200), and the electrode section (300), and providing a passage so that the composite nanostructure injected into the injection section (100) can flow into the reaction section (200) and the electrode section (300). The nanoparticle may have peroxidase-mimicking activity.

[0019] Furthermore, the dual immune diagnostic chip (10) may further include an exhaust section (500) that allows the injected composite nanostructure to be discharged to the outside after moving to the electrode section (300).

[0020] In one embodiment of the present invention, the composite nanostructure may be one in which a nanoparticle and a first bioreceptor are combined. The composite may be called a conjugate or a complex, etc., in which the nanoparticle and the bioreceptor are connected.

[0021] In one embodiment of the present invention, the sample may be the subject of a test to determine whether it contains a target substance. For example, the sample may be a sample obtained from blood, plasma, serum, bone marrow fluid, lymph fluid, saliva, urine, tears, mucosal fluid, amniotic fluid, and various tissues within the body.

[0022] In one embodiment of the present invention, the nanoparticles may have peroxidase-mimicking activity. That is, the nanoparticles can oxidize other substrates while reducing hydrogen peroxide (H2O2) to water (H2O) like peroxidase. For example, the nanoparticles may be one or more selected from the group consisting of Prussian blue nanoparticles, gold-platinum bimetallic nanoparticles, cobalt oxyhydroxide (CoOOH) / carbon black (CB) nanoparticles, and analogs thereof.

[0023] In one embodiment of the present invention, the first bioreceptor may bind to a target substance contained in a sample. For example, the first bioreceptor may be one or more selected from the group consisting of antibodies, enzymes, antigens, aptamers, lectin nucleic acids, proteins, lipids, and glucose hormone receptors. If the first bioreceptor is an antibody, it may be an antibody that specifically binds to a target substance. The target substance may be a target that the user intends to detect within the sample.

[0024] The first bioreceptor may be connected to a nanoparticle. For example, a first bioreceptor composed of an antibody may be connected to a nanoparticle composed of Prussian blue nanoparticles.

[0025] In one embodiment of the present invention, the injection part (100) can receive a composite nanostructure from the outside. To this end, the injection part (100) may have a shape that allows it to receive a composite nanostructure from the outside. For example, the injection part (100) may have a shape with an open top.

[0026] A microfluidic channel (400) may be connected to one side of the injection part (100). Accordingly, a composite nanostructure injected from the outside can flow through the injection part (100) to the microfluidic channel (400).

[0027] In one embodiment of the present invention, the reaction unit (200) may change color depending on the presence or absence of a target substance in the sample, thereby allowing the user to determine whether there is a target substance in the sample. To this end, the reaction unit (200) may have a shape that allows the change in color to be checked from the outside. For example, the reaction unit (200) may have a shape with an open top. Alternatively, a material capable of transmitting light may be located on the top of the reaction unit (200).

[0028] A microfluidic channel section (400) may be connected to one side of the reaction section (200). Accordingly, the composite nanostructure injected through the injection section (100) can flow into the reaction section (200) through the microfluidic channel section (400).

[0029] A color-changing reagent may be applied to the reaction section (200). For example, the reaction section (200) may include paper (210) coated with the color-changing reagent. The paper (210) coated with the color-changing reagent may be located at the bottom of the reaction section (200).

[0030] The reaction section (200) can emit different colors depending on whether the sample contains a target substance or not. For example, consider the case where the nanoparticles are Prussian blue nanoparticles and the first bioreceptor is an antibody. Prussian blue nanoparticles exhibit a deep blue color. Prussian blue nanoparticles exhibit peroxidase-mimicking activity. Prussian blue induces a redox reaction in the presence of TMB (3,3',5,5'-Tetramethylbenzidine) and hydrogen peroxide (H2O2) to produce oxidized TMB and water (H2O). In this case, a color change can be observed with the naked eye. However, when the sample contains a target substance, the aforementioned redox reaction does not occur. This is because the target substance binds to the antibody and blocks the aforementioned reaction. In this case, no color change is observed.

[0031] In one embodiment of the present invention, the color change reagent may include (i) and (ii). (i) may include hydrogen peroxide (H2O2). (ii) may include at least one of ABTS (2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), TMB (3,3',5,5'-Tetramethylbenzidine), AEC (3-Amino-9-Ethylcarbazol), DAB (3,3'-diaminobenzidine tetrahydrochloride salt), OPD (o-phenylenediamine dihydrochloride), luminol, and Amplex Red.

[0032] In one embodiment of the present invention, the electrode portion (300) can generate different electrical signals depending on whether or not the sample contains a target substance. To this end, the electrode portion (300) may have a shape capable of generating an electrical signal.

[0033] A microfluidic channel (400) may be connected to one side of the electrode portion (300). Accordingly, a composite nanostructure injected from the outside can flow into the electrode portion (300) through the microfluidic channel (400). More specifically, the microfluidic channel (400) is connected to the electrode portion (300) chamber included in the electrode portion (300), and the composite nanostructure can flow into the electrode portion (300) chamber through the microfluidic channel (400).

[0034] A second bioreceptor capable of binding to a target substance may be immobilized on the electrode portion (300). The second bioreceptor may be one or more selected from the group consisting of antibodies, enzymes, antigens, aptamers, lectin nucleic acids, proteins, lipids, and glucose hormone receptors.

[0035] The electrode portion (300) may be a triode (Fig. 8). The triode may include a reference electrode, a working electrode, and a counter electrode. The reference electrode may be made of AG or Ag / AgCl paste. The working electrode may be made of Au or Carbon. The counter electrode may be made of Pt or Carbon.

[0036] The orientation of the electrode portion (300) can vary. For example, the electrode portion (300) may be positioned parallel to or perpendicular to the flow direction of the composite nanostructure, as shown in Type 1 to Type 3 of FIG. 8. Therefore, the dual immunodiagnostic chip (10) may have the shape of FIG. 2, but may also have a shape in which the orientation of the electrode portion is horizontal, as shown in FIG. 9.

[0037] The electrode portion (300) may have a protruding shape (Fig. 8 (a)). That is, one side of the electrode portion (300) may protrude outside the dual immune diagnostic chip (10). A connector capable of interlocking with an external electronic device may be connected to one end of the electrode portion (300) that protrudes outside.

[0038] The electrode portion (300) may have an insertable shape (Fig. 8 (b)). That is, one side of the electrode portion (300) may be inserted inside the dual immune diagnostic chip (10). A contact portion capable of contacting an external electronic device may be connected to one end of the inserted electrode portion (300).

[0039] The electrode portion (300) can generate different electrical signals depending on whether the sample contains a target substance or not. For example, consider the case where the nanoparticle is a Prussian blue nanoparticle and the first bioreceptor and second bioreceptor are antibodies (Figs. 11 and 12). When the sample does not contain a target substance, the antibody (second bioreceptor) fixed to the electrode portion (300) cannot bind to the composite nanostructure. In this case, there is no significant change in the current (electrical signal). Conversely, when the sample contains a target substance, the antibody (second bioreceptor) fixed to the electrode portion (300) can bind to the composite nanostructure. This is because the target substance is bound to the antibody (first bioreceptor) connected to the Prussian blue nanoparticle. In this case, a change in the current (electrical signal) occurs. By measuring the change in the current (electrical signal) in this way, it is possible to detect whether the target substance is present in the sample.

[0040] In one embodiment of the present invention, the microfluidic channel portion (400) is connected to the injection portion (100), the reaction portion (200), and the electrode portion (300), and may provide a passage so that a composite nanostructure injected into the injection portion (100) can flow into the reaction portion (200) and the electrode portion (300).

[0041] In one embodiment of the present invention, the absorption pad may be located at the bottom of the microchannel fluid section. The absorption pad (410) can control the fluid flow of the composite nanostructure flowing inside the microfluidic channel section (400).

[0042] The absorbent pad (410) may be composed of at least one of the group consisting of porous absorbent paper and fabric materials.

[0044] The dual immune diagnostic chip (10) may be composed of three layers (Fig. 3). The thickness of the three layers may be 300 μm to 1000 μm. The three layers may be composed of biocompatible hard plastic materials. For example, the three layers may be composed of materials such as polycarbonate and polyester.

[0045] Each of the three layers can be called the first layer, the second layer, and the third layer.

[0046] The first layer may be located at the very bottom. The first layer may form the bottom surface of the microfluidic channel section (400). The first layer may have a hydrophilic coating at the required location. For example, the second layer may have a hydrophilic coating at the location of the injection section (100), microfluidic channel section (400), and electrode section (300) chambers where the composite nanostructure enters and exits.

[0047] The second layer may be located between the first layer and the third layer. In the second layer, an empty space may be formed at the location where the injection section (100), reaction section (200), microfluidic channel section (400), and electrode section (300) chambers are located. Furthermore, an empty space may be formed in the second layer at the location where the discharge section (500) is located.

[0048] The third layer may be located at the very top. A void space may be formed in the third layer at a location corresponding to the injection part (100). Furthermore, a void space may be formed in the third layer at a location where the discharge part (500) is located. Accordingly, a composite nanostructure may be injected or discharged through the void space formed in the third layer.

[0049] The absorbent pad (410) can be placed in the empty space at the location of the microfluidic channel section (400) in the second layer. That is, as shown in FIG. 3, the absorbent pad (410) can be located at the location of the microfluidic channel section (400) at the top of the first layer.

[0050] The absorbent pad (410) may be thinner than the thickness of the second layer. In one embodiment, if the thickness of the second layer is 300 to 400 μm, the thickness of the absorbent pad (410) may be 150 μm. Accordingly, a flow space may be created on the upper part of the microfluidic channel section (400).

[0051] The paper (210) coated with a discoloration reagent can be placed in the empty space at the location of the reaction section (200) in the second layer. That is, as shown in FIG. 3, the absorption pad can be placed at the location of the reaction section (200) on top of the first layer and the absorption pad (410).

[0053] Another aspect of the present invention provides a dual detection method for target substances using a dual immunodiagnostic chip.

[0054] A method for detecting a target substance using a dual immunodiagnostic chip comprises: a step of injecting a composite nanostructure, in which a sample, a nanoparticle, and a first bioreceptor are combined, into an injection part (100); and a step of detecting whether a target substance is present in the sample based on the result of measuring (a) or (b). (a) may be a color change of the reaction part (200) of the dual immunodiagnostic chip. (b) may be a change in the electrical signal of the electrode part (300) of the dual immunodiagnostic chip. Furthermore, the method for detecting a target substance using a dual immunodiagnostic chip may further include a step of washing the inside of the dual immunodiagnostic chip.

[0055] Furthermore, the method for detecting target substances using a dual immunodiagnostic chip may further include a step of inducing a composite nanostructure to flow into a reaction section (200) and an electrode section (300) using a magnetic material after injecting the composite nanostructure. For example, as shown in FIG. 3, a magnetic material (magnet) may be placed at the electrode section (300) to allow the composite nanostructure to flow into the electrode section (300).

[0056] The detection step may be a step of detecting whether a target substance is present in the sample by simultaneously measuring (a) and (b). As described above, if the target substance is present in the sample, an oxidation-reduction reaction does not occur, so a color change in the reaction section (200) is not observed, but a change in current in the electrode section (300) can be observed. Conversely, if the target substance is not present in the sample, an oxidation-reduction reaction occurs, so a color change in the reaction section (200) is observed, but a change in current in the electrode section (300) is not observed. In this way, by simultaneously measuring the color change and electrical signal with a single sample injection, the detection of the target substance can be performed more accurately and quickly. Furthermore, the target substance can be detected by observing the color change and electrical signal change with a small amount of sample.

[0057] The detection step may be a step of detecting whether a target substance is present in the sample by comparing the measured result with a preset value. Explanation of the symbols

[0058] 100 : Injection part 200 : Reaction part 210: Paper coated with discoloration reagent 300 : Electrode part 400: Microfluidic channel section 410: Absorbent pad 500 : Discharge part

Claims

Claim 1 An injection section (100) into which a composite nanostructure, comprising a sample, a nano particle, and a first bioreceptor, is injected; a reaction section (200) coated with a color-changing reagent that reacts with the composite nanostructure to emit color; an electrode section (300) in which a second bioreceptor capable of binding to a target substance is fixed and generates an electrical signal; a microfluidic channel section (400) connected to the injection section (100), the reaction section (200), and the electrode section (300), and providing a passage so that the composite nanostructure injected into the injection section (100) can flow into the reaction section (200) and the electrode section (300); and an absorption pad (410) located at the bottom of the microfluidic channel section (400) and controlling the fluid flow of the composite nanostructure flowing inside the microfluidic channel section (400). A dual immunodiagnostic chip comprising, wherein the nanoparticles have peroxidase-mimicking activity. Claim 2 A dual immunodiagnostic chip according to claim 1, wherein the nanoparticles are one or more selected from the group consisting of Prussian Blue nanoparticles, gold-platinum bimetallic nanoparticles, and cobalt oxyhydroxide (CoOOH) / carbon black (CB) nanoparticles. Claim 3 A dual immunodiagnostic chip according to claim 1, wherein the first bioreceptor is one or more selected from the group consisting of antibodies, enzymes, antigens, aptamers, lectin nucleic acids, proteins, lipids, and glucose hormone receptors. Claim 4 A dual immunodiagnostic chip according to claim 1, wherein the second bioreceptor is one or more selected from the group consisting of antibodies, enzymes, antigens, aptamers, lectin nucleic acids, proteins, lipids, and glucose hormone receptors. Claim 5 A dual immunodiagnostic chip according to claim 1, wherein the color change reagent comprises at least one of ABTS (2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), TMB (3,3',5,5'-Tetramethylbenzidine), AEC (3-Amino-9-Ethylcarbazol), DAB ((3,3'-diaminobenzidine tetrahydrochloride salt), OPD (o-phenylenediamine dihydrochloride), luminol, and Amplex Red. Claim 6 A dual immune diagnostic chip according to claim 1, wherein one side of the electrode portion (300) protrudes outside the dual immune diagnostic chip, and further comprising a connector connected to one end of the electrode portion (300) protruding outside so that the dual immune diagnostic chip can be coupled with an external electronic device. Claim 7 A dual immune diagnostic chip according to claim 1, wherein one side of the electrode portion (300) is inserted into the dual immune diagnostic chip, and further comprising a contact portion located at one end of the inserted electrode portion (300) so that the dual immune diagnostic chip can contact an external electronic device. Claim 8 delete Claim 9 In claim 1, the absorbent pad (410) is a dual immunodiagnostic chip selected from at least one of the group consisting of porous absorbent paper and fabric material. Claim 10 A dual immune diagnostic chip according to claim 1, further comprising an exhaust portion (500) that allows the composite nanostructure to be discharged to the outside after moving to the electrode portion (300). Claim 11 A method for dual detection of a target substance using a dual immunodiagnostic chip according to claim 1, comprising: a step of injecting a composite nanostructure, in which a sample, a nanoparticle, and a first bioreceptor are combined, into an injection part (100); and a step of detecting whether a target substance is present in the sample based on the result of measuring (a) or (b) below. (a) a color change of the reaction part (200) of the dual immunodiagnostic chip; (b) a change in the electrical signal of the electrode part (300) of the dual immunodiagnostic chip. Claim 12 A dual detection method for target substances using a dual immunodiagnostic chip, wherein the detecting step is a step of simultaneously measuring (a) and (b) to detect whether a target substance is present in the sample. Claim 13 A method for detecting a target substance using a dual immunodiagnostic chip, wherein the detecting step comprises: a step of detecting whether the target substance is present in the sample by comparing the measured result with a preset value in the above-mentioned detection step. Claim 14 A method for detecting a target substance using a dual immunodiagnostic chip, wherein, in claim 11, the method further comprises the step of washing the interior of the dual immunodiagnostic chip. Claim 15 A method for detecting a target substance using a dual immunodiagnostic chip, wherein, in claim 11, after the step of injecting the composite nanostructure, the method further comprises the step of using a magnetic material to induce the injected composite nanostructure to flow to the reaction section (200) and the electrode section (300).

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