Dual immunodiagnostic chip

Abstract

The present invention provides a dual immunodiagnostic chip. The dual immunodiagnostic chip can be used to observe changes in color and electrical signal, through a single sample injection. Thus, the presence or absence of a target material can be rapidly and accurately detected. The dual immunodiagnostic chip of the present invention comprises: an injection unit (100) into which a composite nanostructure in which a sample, nanoparticles, and a primary bioreceptor are conjugated; a reaction unit (200) coated with a chromogenic reagent that develops color by reacting with the composite nanostructure; an electrode unit (300) on which a secondary bioreceptor capable of binding to a target material is immobilized and which generates an electrical signal; and a microfluidic channel unit (400) which is connected to the injection unit (100), the reaction unit (200), and the electrode unit (300) and provides a passage through which the composite nanostructure injected into the injection unit (100) can flow into the reaction unit (200) and the electrode unit (300).

Description

Dual immune diagnostic chip

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

[0002] Microfluidics-based technology deals with handling small volumes of fluid. Microfluidics is utilized in diverse fields, including biotechnology, medicine, the environment, and physics. Recently, technologies utilizing microfluidics to detect the presence of target substances in samples are being developed. Specifically, immunodiagnostic chips, which can detect cancer or viruses, are being utilized in combination with composite nanostructure technology that combines antibodies with functional nanoparticles. These immunodiagnostic chips operate by activating changes in optical or electrochemical signals depending on the presence of target substances, enabling the detection of cancer, viruses, and genetic material.

[0003] [Prior Art Literature]

[0004] [Patent Document]

[0005] Korean Patent No. 10-1188172

[0006] One aspect is to provide a dual immune diagnostic chip.

[0007] Another aspect is to provide a method for detecting target substances using a dual immune diagnostic chip.

[0008] The dual immune diagnostic chip includes an injection unit (100) into which a composite nanostructure in which a sample, nanoparticles, and a first bioreceptor are combined is injected; a reaction unit (200) on which a color-changing reagent that reacts with the composite nanostructure to emit color is applied; an electrode unit (300) on which a second bioreceptor capable of binding to a target substance is fixed and which generates an electric signal; and a microfluidic channel unit (400) connected to the injection unit (100), the reaction unit (200), and the electrode unit (300), and providing a passage through which the composite nanostructure injected into the injection unit (100) can flow to the reaction unit (200) and the electrode unit (300).

[0009] A method for detecting a target substance using a dual immune diagnostic chip includes a step of injecting a composite nanostructure in which a sample, nanoparticles, and a first bioreceptor are combined into an injection unit (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) Color change of a reaction unit (200) of a dual immune diagnostic chip (b) Electrical signal change of an electrode unit (300) of a dual immune diagnostic chip

[0010] The technology described below can be used to detect the presence of a target substance in a sample.

[0011] Using the technology described below, the presence of a target substance within a sample can be detected by observing color and electrical signal changes with a single sample introduction. This allows for rapid and accurate detection of the presence of a target substance within a sample, even with small sample volumes.

[0012] Figure 1 is an overall overview of the dual immune diagnostic chip.

[0013] Figure 2 is a perspective view of one embodiment of a dual immune diagnostic chip (10).

[0014] Figure 3 is an exploded view of one embodiment of a dual immune diagnostic chip (10).

[0015] Figure 4 is an embodiment of the first layer of a dual immune diagnostic chip (10).

[0016] Figure 5 is an embodiment of the second layer of a dual immune diagnostic chip (10).

[0017] Figure 6 is an embodiment of the third layer of a dual immune diagnostic chip (10).

[0018] Figure 7 is an example of an electrode part (300).

[0019] Figure 8 is an example of an electrode part (300).

[0020] Figure 9 is a plan view of one embodiment of a dual immune diagnostic chip (10).

[0021] Fig. 10 is an example of a reaction that may occur in the reaction section (200). Fig. 10 is an enlarged view of a portion of Fig. 1.

[0022] Fig. 11 is an example of a reaction that may occur in the electrode section (300). Fig. 11 is an enlarged view of a portion of Fig. 1.

[0023] Figure 12 is an example of a change in an electric signal that may occur in an electrode section (300).

[0024] Hereinafter, embodiments are described in detail with reference to exemplary drawings. When designating components in each drawing, it should be noted that, where possible, identical components are given identical reference numbers, even if they appear in different drawings. Furthermore, when describing embodiments, detailed descriptions of related known configurations or functions will be omitted if they are deemed to hinder understanding of the embodiments.

[0025] The inventors have developed a dual immunodiagnostic chip (Fig. 1). They have also developed a method for detecting target substances using this chip. The dual immunodiagnostic chip can simultaneously perform optical and electrochemical analyses with a single sample input. This allows for rapid and accurate detection of target substances even with a small sample volume.

[0026] The present invention is described in detail below.

[0027] One aspect of the present invention provides a dual immune diagnostic chip (10).

[0028] 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 embodiment of a first layer of a dual immune diagnostic chip (10). FIG. 5 is an embodiment of a second layer of a dual immune diagnostic chip (10). FIG. 6 is an embodiment of a third layer of a dual immune diagnostic chip (10). FIG. 7 is an embodiment of an electrode unit (300). FIG. 8 is an embodiment of an electrode unit (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 unit (200). FIG. 11 is an example of a reaction that may occur in an electrode unit (300). FIG. 12 is an example of a change in an electrical signal that may occur in an electrode unit (300).

[0029] A dual immune diagnostic chip (10) includes an injection unit (100) into which a sample, a nano particle, and a first bioreceptor are combined and a composite nano structure are injected; a reaction unit (200) on which a color-changing reagent that reacts with the composite nano structure to emit color is applied; an electrode unit (300) on which a second bioreceptor capable of binding to a target substance is fixed and which generates an electric signal; and a microfluidic channel unit (400) connected to the injection unit (100), the reaction unit (200), and the electrode unit (300), which provides a passage through which the composite nano structure injected into the injection unit (100) can flow to the reaction unit (200) and the electrode unit (300). The nano particle may have peroxidase-mimicking activity.

[0030] Furthermore, the dual immune diagnostic chip (10) may further include an exhaust section (500) that allows the injected complex nanostructure to move to the electrode section (300) and then be exhausted to the outside.

[0031] In one embodiment of the present invention, the composite nanostructure may be a combination of a nanoparticle and a first bioreceptor. The composite may be referred to as a conjugate or complex, as the nanoparticle and the bioreceptor are linked.

[0032] In one embodiment of the present invention, a sample may be the subject of a test to determine whether it contains a target substance. For example, the sample may be blood, plasma, serum, bone marrow fluid, lymph, saliva, urine, tears, mucous membrane fluid, amniotic fluid, or samples obtained from various tissues within the body.

[0033] In one embodiment of the present invention, the nanoparticles may have peroxidase-mimetic activity. That is, the nanoparticles can reduce hydrogen peroxide (H2O2) to water (H2O) like a peroxidase, while oxidizing other substrates. 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.

[0034] In one embodiment of the present invention, the first bioreceptor can 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 sugar hormone receptors. If the first bioreceptor is an antibody, it may be an antibody that specifically binds to the target substance. The target substance may be a substance that the user wishes to detect in the sample.

[0035] The first bioreceptor may be linked to a nanoparticle. For example, the first bioreceptor, which is comprised of an antibody, may be linked to a nanoparticle comprised of Prussian Blue nanoparticles.

[0036] In one embodiment of the present invention, the injection unit (100) can receive a composite nanostructure from the outside. To this end, the injection unit (100) may have a shape that allows the composite nanostructure to be injected from the outside. For example, the injection unit (100) may have a shape with an open top.

[0037] A microfluidic channel section (400) may be connected to one side of the injection section (100). Accordingly, a composite nanostructure injected from the outside can flow into the microfluidic channel section (400) through the injection section (100).

[0038] 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 a sample, thereby allowing a user to determine whether the target substance is present in the sample. To this end, the reaction unit (200) may have a shape that allows the color change to be observed from the outside. For example, the reaction unit (200) may have a shape with an open top. Alternatively, a material that can transmit light may be positioned on the top of the reaction unit (200).

[0039] 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) may flow into the reaction section (200) through the microfluidic channel section (400).

[0040] A color-changing reagent may be applied to the reaction unit (200). For example, the reaction unit (200) may include paper (210) to which the color-changing reagent is applied. The paper (210) to which the color-changing reagent is applied may be positioned at the bottom of the reaction unit (200).

[0041] The reaction unit (200) can emit different colors depending on whether or not the sample contains a target substance. As an example, let's consider a 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-mimetic 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, if the sample contains a target substance, the aforementioned redox reaction does not occur. This is because the target substance binds to the antibody, blocking the aforementioned reaction. In this case, no color change is observed.

[0042] In one embodiment of the present invention, the color-changing 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.

[0043] In one embodiment of the present invention, the electrode unit (300) can generate different electrical signals depending on whether a target substance is contained in the sample. To this end, the electrode unit (300) can have a shape capable of generating an electrical signal.

[0044] A microfluidic channel portion (400) may be connected to one side of the electrode portion (300). Accordingly, a composite nanostructure injected from the outside may flow into the electrode portion (300) through the microfluidic channel portion (400). More specifically, the microfluidic channel portion (400) is connected to the electrode portion (300) chamber included in the electrode portion (300), and the composite nanostructure may flow into the electrode portion (300) chamber through the microfluidic channel portion (400).

[0045] A second bioreceptor capable of binding to a target substance may be fixed to 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 sugar hormone receptors.

[0046] The electrode unit (300) may be a three-pole electrode (Fig. 8). The three-pole electrode may include a reference electrode, a working electrode, and a counter electrode. The reference electrode may be made of AG, Ag / AgCl paste. The working electrode may be made of Au, Carbon. The counter electrode may be made of Pt, Carbon.

[0047] The direction of the electrode portion (300) may vary. For example, the electrode portion (300) may be positioned parallel to or perpendicular to the direction of movement (Flow) of the composite nanostructure, as in Type 1 to Type 3 of FIG. 8. Accordingly, the dual immune diagnostic chip (10) may have the shape of FIG. 2, but may also have a shape in which the direction of the electrode portion is horizontal, as in FIG. 9.

[0048] 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 that can be linked to an external electronic device may be connected to one end of the electrode portion (300) protruding outside.

[0049] The electrode portion (300) may have an insertion shape (Fig. 8 (b)). That is, one side of the electrode portion (300) may be inserted into the dual immune diagnostic chip (10). A contact portion that can make contact with an external electronic device may be connected to one end of the inserted electrode portion (300).

[0050] The electrode unit (300) can generate different electrical signals when the sample contains a target substance and when it does not. For example, consider the case where the nanoparticles are Prussian blue nanoparticles and the first bioreceptor and the second bioreceptor are antibodies (Figs. 11 and 12). When the sample does not contain a target substance, the antibody (second bioreceptor) immobilized on the electrode unit (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) immobilized on the electrode unit (300) can bind to the composite nanostructure. This is because the target substance is bound to the antibody (first bioreceptor) linked to the Prussian blue nanoparticle. In this case, a change in the current (electrical signal) occurs. In this way, by measuring the change in the current (electrical signal), the presence of the target substance in the sample can be detected.

[0051] In one embodiment of the present invention, the microfluidic channel section (400) is connected to the injection section (100), the reaction section (200), and the electrode section (300), and can provide a passage through which the composite nanostructure injected into the injection section (100) can flow to the reaction section (200) and the electrode section (300).

[0052] In one embodiment of the present invention, an absorbent pad may be positioned at the bottom of the microfluidic channel portion. The absorbent pad (410) may control the fluid flow of the composite nanostructure flowing within the microfluidic channel portion (400).

[0053] The absorbent pad (410) may be composed of at least one of a group consisting of porous absorbent paper and fabric materials.

[0054] The dual immune diagnostic chip (10) may be composed of three layers (Fig. 3). The thickness of the three layers may range from 300 μm to 1000 μm. The three layers may be composed of a biocompatible hard plastic material. For example, the three layers may be composed of materials such as polycarbonate or polyester.

[0055] The three individual layers can be referred to as the first layer, the second layer, and the third layer.

[0056] The first layer may be positioned at the bottom. The first layer may form the bottom surface of the microfluidic channel portion (400). The first layer may have a hydrophilic coating at required locations. For example, the second layer may have a hydrophilic coating at the injection portion (100), the microfluidic channel portion (400), and the electrode portion (300) chamber locations through which the composite nanostructure enters and exits.

[0057] The second layer may be positioned between the first layer and the third layer. The second layer may have an empty space formed at the location where the injection unit (100), reaction unit (200), microfluidic channel unit (400), and electrode unit (300) chambers are located. Furthermore, the second layer may have an empty space formed at the location where the discharge unit (500) is located.

[0058] The third layer may be positioned at the top. A void may be formed in the third layer at a location corresponding to the injection port (100). Furthermore, a void may be formed in the third layer at a location corresponding to the discharge port (500). Accordingly, the composite nanostructure may be injected or discharged through the void formed in the third layer.

[0059] The absorption pad (410) can be placed in the empty space at the position of the microfluidic channel portion (400) in the second layer. That is, as shown in FIG. 3, the absorption pad (410) can be positioned at the position of the microfluidic channel portion (400) at the top of the first layer.

[0060] 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. This may create a flow space above the microfluidic channel portion (400).

[0061] The paper (210) to which the discoloration reagent is applied can be placed in the empty space at the position of the reaction part (200) in the second layer. That is, as shown in FIG. 3, the absorption pad can be positioned at the position of the reaction part (200) on top of the first layer and the absorption pad (410).

[0062] Another aspect of the present invention provides a method for dual detection of a target substance using a dual immune diagnostic chip.

[0063] A method for detecting a target substance using a dual immune diagnostic chip includes the steps of: injecting a composite nanostructure in which a sample, nanoparticles, and a first bioreceptor are combined into an injection unit (100); and 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 in a reaction unit (200) of the dual immune diagnostic chip. (b) may be a change in an electric signal in an electrode unit (300) of the dual immune diagnostic chip. Furthermore, the method for detecting a target substance using a dual immune diagnostic chip may further include a step of washing the inside of the dual immune diagnostic chip.

[0064] Furthermore, the method for detecting a target substance using a dual immune diagnostic chip may further include a step of injecting a composite nanostructure and then inducing the injected composite nanostructure to flow to the reaction unit (200) and the electrode unit (300) using a magnetic material. For example, as shown in FIG. 3, a magnetic material (magnet) may be positioned on the electrode unit (300) to allow the composite nanostructure to flow into the electrode unit (300).

[0065] The detecting step may be a step of simultaneously measuring (a) and (b) to detect whether a target substance is present in the sample. As described above, if the target substance is present in the sample, a redox reaction does not occur, so that no color change in the reaction unit (200) is observed, but a change in the current of the electrode unit (300) can be observed. Conversely, if the target substance is not present in the sample, a redox reaction occurs, so that a color change in the reaction unit (200) is observed, but a change in the current of the electrode unit (300) is not observed. In this way, by simultaneously measuring color change and electric signal with a single sample injection, target substance detection can be made more accurate and rapid. In addition, target substances can be detected by observing color change and electric signal change with a small amount of sample.

[0066] The detection step may be a step of comparing the measured result with a preset value to detect whether a target substance is present in the sample.

[0067] [Description of drawing symbols]

[0068] 100: Injection part

[0069] 200: Reaction section

[0070] 210: Paper coated with discoloration agent

[0071] 300: Electrode section

[0072] 400: Microfluidic channel section

[0073] 410: Absorbent pad

[0074] 500: exhaust

Claims

1. An injection unit (100) into which a composite nanostructure comprising a sample, nano particles, and a first bioreceptor is injected; A reaction section (200) to which a color-changing reagent that reacts with a composite nanostructure to emit color is applied; An electrode part (300) on which a second bioreceptor capable of binding to a target substance is fixed and which generates an electric signal; and A microfluidic channel section (400) connected to the injection section (100), reaction section (200) and electrode section (300), and providing a passage so that the composite nanostructure injected into the injection section (100) can flow to the reaction section (200) and electrode section (300); including, The above nanoparticles are a dual immunodiagnostic chip having peroxidase-mimicking activity.

2. In paragraph 1, A dual immunodiagnostic chip, wherein the nanoparticles are at least one selected from the group consisting of Prussian Blue nanoparticles, gold-platinum bimetallic nanoparticles, cobalt oxyhydroxide (CoOOH) / carbon black (CB) nanoparticles, and analogs thereof.

3. In paragraph 1, A dual immunodiagnostic chip, wherein the first bioreceptor is at least one selected from the group consisting of antibodies, enzymes, antigens, aptamers, lectin nucleic acids, proteins, lipids, and sugar hormone receptors.

4. In paragraph 1, A dual immunodiagnostic chip, wherein the second bioreceptor is at least one selected from the group consisting of antibodies, enzymes, antigens, aptamers, lectin nucleic acids, proteins, lipids, and sugar hormone receptors.

5. In paragraph 1, A dual immunodiagnostic chip, wherein the color-changing 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.

6. In paragraph 1, One side of the above electrode portion (300) protrudes outside the dual immune diagnostic chip, A dual immune diagnostic chip further comprising a connector connected to one end of the electrode portion (300) protruding outwardly so that the dual immune diagnostic chip can be linked with an external electronic device.

7. In paragraph 1, One side of the above electrode part (300) is inserted inside the dual immune diagnostic chip, A dual immune diagnostic chip further comprising a contact portion located at one end of the inserted electrode portion (300) through which the dual immune diagnostic chip can contact an external electronic device.

8. In paragraph 1, A dual immune diagnostic chip further comprising an absorption pad (410) positioned at the bottom of the microfluidic channel section (400) and controlling the fluid flow of the complex nanostructure flowing inside the microfluidic channel section (400).

9. In paragraph 8, The above microfluidic channel portion (400) is a dual immune diagnostic chip made of at least one selected from the group consisting of porous absorbent paper and fabric materials.

10. In paragraph 1, A dual immune diagnostic chip further comprising an exhaust section (500) that allows the above-mentioned composite nanostructure to move to the electrode section (300) and then be exhausted to the outside.

11. In the method for dual detection of a target substance using the dual immune diagnostic chip of paragraph 1, A step of injecting a composite nanostructure in which a sample, nanoparticles, and a first bioreceptor are combined into the injection unit (100); and A method for detecting a target substance using a dual immune diagnostic chip, comprising: a step of detecting whether a target substance is present in the sample based on the result of measuring (a) or (b). (a) Color change of the reaction part (200) of the dual immune diagnostic chip (b) Changes in electrical signals of the electrode portion (300) of the dual immune diagnostic chip 12. In paragraph 11, A method for dual detection of a target substance using a dual immune diagnostic 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.

13. In paragraph 11, A method for detecting a target substance using a dual immune diagnostic chip, wherein the detecting step comprises a step of comparing the measured result with a preset value to detect whether the target substance is present in the sample.

14. In paragraph 11, A method for detecting a target substance using a dual immune diagnostic chip, further comprising a step of washing the interior of the dual immune diagnostic chip.

15. In paragraph 11, A method for detecting a target substance using a dual immune diagnostic chip, further comprising, after the step of injecting the above-mentioned composite nanostructure, a step of inducing the injected composite nanostructure to flow to the reaction unit (200) and electrode unit (300) using a magnetic material.

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