KR102066268B1 - A kit for detecting target materials comprising image sensors and magnets - Google Patents

Abstract

The present invention relates to a kit for detecting a target material including two or more particles and a target material detection unit which can easily check the presence or absence of a target material. It is possible to form an asymmetric complex, which can be easily identified and analyzed to efficiently detect a plurality of targets.

Description

A kit for detecting target materials comprising image sensors and magnets}

The present invention relates to a kit for detecting a target substance including two or more particles and a target substance detection unit which can easily check the presence or absence of a target substance.

Techniques for detecting or isolating target substances in samples are used in a variety of fields, including disease diagnosis, food testing, and environmental monitoring. Conventional detection methods include immunoassays, and radioimmunoassays, enzyme immunoassays, fluorescent antibodies, chemiluminescent immunoassays according to the detection principle and method. Recently, there have been attempts to increase detection sensitivity by binding various types of nanoparticles such as gold and silica to the surface of antibodies or enzymes.

As a method of detecting or separating a specific target material from a biological sample, a method using silica, glass fiber, anion exchange resin or magnetic beads is known. According to the method of using magnetic beads among them, magnetic beads having a probe capable of binding to the target biomaterial on the surface thereof are introduced into the sample solution to capture the target biomaterial, and the magnetic beads and the target biomaterial are separated again. Extract material separately. As described above, a method of detecting or separating a target biomaterial using magnetic beads has been widely used to detect or isolate substances such as cells, proteins, nucleic acids, and the like (Non-Patent Document 1).

An image sensor refers to a device or an electronic component that detects subject information and converts it into an electrical image signal. The image sensor may be broadly classified into an image pickup tube and a solid state image sensor. The former includes a vidicon, a plumbicon, and the latter includes a complementary metal-oxide semiconductor (CMOS) and a charged coupled device (CCD). Used for

(Non-Patent Document 1) 1.BioChip J., Vol. 8, 1 (2014), 1 ~ 7

One object of the present invention is a magnetic field generating unit for generating a magnetic field; An image sensor connected in line with one side of the magnetic field generator; And a thin film attached to a surface of the image sensor; And different binding molecules that specifically bind to different recognition sites respectively present in one or more types of target materials, wherein the binding molecules include magnetic particles or particles, and one type of target material includes magnetic particles and The different binding molecules specifically bind to the target material so that one kind of particles are contained, the target material is collected in the thin film by the magnetic field generating unit, and the collection is performed without passing through the microfluidic channel. The image sensor is a complementary metal oxide semiconductor image sensor, and provides a kit for detecting a target material, wherein the collection and imaging of the target material by the image sensor are performed at the same time.

One aspect of the invention the magnetic field generating unit for generating a magnetic field; An image sensor connected in line with one side of the magnetic field generator; And a thin film attached to a surface of the image sensor; And different binding molecules that specifically bind to different recognition sites respectively present in one or more types of target materials, wherein the binding molecules include magnetic particles or particles, and one type of target material includes magnetic particles and The different binding molecules specifically bind to the target material so that one kind of particles are contained, the target material is collected in the thin film by the magnetic field generating unit, and the collection is performed without passing through the microfluidic channel. The image sensor is a complementary metal-oxide semiconductor image sensor, and provides a kit for detecting a target material, wherein the collection and imaging of the target material by the image sensor are performed at the same time.

According to an embodiment of the present invention, the magnetic field generating unit included in the kit may be selected from the group consisting of magnets, solenoids and superconducting magnets, and is preferably a magnet.

As used herein, the term "magnetic field" refers to a range in which a magnetic force acts around a magnet, an electric current, a changing electric field, and the like. It means the configuration to create. A material that maintains its own magnetic field without maintaining an external magnetic field is called a magnet (ferromagnetic material). An electro-magnet is a magnet that is magnetized when an electric current flows, such as a solenoid or a superconducting magnet.

As used herein, the term "image sensor" refers to a device or an electronic component that detects subject information and converts it into an electric image signal, and can be broadly divided into an imaging tube and a solid state image sensor, and a CMOS and CCD image. The sensor belongs to a solid image sensor. In the kit, the image sensor is preferably a complementary metal oxide semiconductor (CMOS) image sensor.

According to one embodiment of the invention, the kit may further include a display unit for displaying a signal of the image sensor as an image, the display unit may be used a display device or the like used in the art.

According to one embodiment of the present invention, the thin film may be selected from the group consisting of a metal thin film, a compound thin film, a semiconductor thin film and an insulating thin film, but is not limited thereto. Preferably, the thin film may be an insulating thin film, and most preferably, the thin film may be a glass plate.

As used herein, the term "thin film" refers to a thin film that prevents direct contact between the image sensor and the sample.

According to one embodiment of the invention, the particles in the kit may be used regardless of the material of the particles, such as gold particles, metal particles such as silver particles, silica particles and polymer particles, preferably polystyrene particles. In addition, the kind of particles in the kit may be two or more, it is most preferably two kinds. In the case of two kinds of particles, the materials of the particles are preferably different from each other, and one of the two particles is most preferably a magnetic particle.

According to one embodiment, it is preferable that the particles have different sizes, and when one particle size is nanometer scale, the other particle may be nanometer or micrometer size larger than that. In addition, it is preferable that the particles having small size are magnetic particles, and the size thereof is preferably in the range of 100 nm to 3 μm.

Unlike bulk metals, metal nanoparticles exhibit a strong absorption phenomenon called surface plasmon resonance band in the visible region, and the absorption wavelength is known to change according to the size and shape of the particles. Therefore, by varying the size and shape of the metal nanoparticles used according to the target material, and confirming the image while changing the wavelength range of the irradiated light can be detected for a plurality of target materials.

In addition, the particles may have a variety of shapes, such as spherical, rod-shaped, linear or polyhedral, and the shape of the two particles may be different from each other, but is preferably spherical. If different types of particles are used for each target material, the presence of a plurality of target materials can be confirmed by checking an image of the particles captured in the thin film.

As used herein, the term "binding molecules" is linked to the surface of the particles through chemical bonding, and can specifically bind to the target material, thereby mediating specific binding of the particles and the target material. It means a molecule.

According to one embodiment of the invention, the binding molecule may be any molecule that can specifically bind to the target material, and may be selected from the group consisting of polypeptides, polynucleotides and lipids, but is not limited thereto. . The binding molecule can be, for example, an enzyme, antibody, ligand, aptamer or micro RNA. Preferably, the binding molecule may be an antibody or aptamer.

As used herein, the term "specifically binding" has the same meaning as is commonly known to those skilled in the art, and the enzyme and the enzyme, This means that molecules such as aptamers and ligands bind to the target with high affinity and specificity.

The kit of the present invention uses two or more kinds of particles having different sizes and binding molecules having different recognition sites. Since the binding molecules recognize different sites of the target material, the target material can be detected with high accuracy. In addition, simultaneous detection of a plurality of target substances is possible as long as it contains a binding molecule specifically recognized for each target substance and a sufficient amount of particles capable of binding to the binding molecule.

A method of detecting a plurality of targets using different binding molecules for each target is shown in FIG. 1.

The present invention is a kit for detecting a target substance including a target substance detection unit and two or more particles, and an example of the kit is shown in FIG. 2. The target material detection unit 100 includes a magnetic field generating unit 110 for generating a magnetic field; An image sensor 120 connected in line with one side of the magnetic field generator; And a thin film 130 attached to a surface of the image sensor.

Figure 3 shows the principle of detecting the target material using the image sensor included in the kit of the present invention. When the target material is present in the sample, the magnetic particle-target material-polystyrene particle complex is formed by two kinds of particles, and the magnetic particles adhere to the thin film on the surface of the image sensor by the magnetic field. Thereafter, the existence of the target material may be confirmed by observing the presence of polystyrene particles on the surface of the thin film. Since the polystyrene particles are larger in size than the magnetic particles, the polystyrene particles can be easily identified, and when the polystyrene particles are present on the surface of the thin film, the polystyrene particle-target material-magnetic particle composite may be formed.

Using the kit of the present invention, it is possible to form asymmetric complexes between particles using different binding molecules with respect to the target material, which can be easily identified and analyzed to efficiently detect a plurality of target materials.

1 is a view schematically showing the principle of detecting a plurality of target substances using different kinds of binding molecules.
Figure 2 is a schematic diagram showing the configuration of the target material detection unit in the target material detection kit of the present invention.
3 is a view showing a method for identifying a protein-antibody-particle complex using an image sensor.
4 is a diagram showing an image of a protein-antibody-particle complex using an image sensor and a magnet.
5 is a graph showing the number of polystyrene particles identified in an image photographed using an image sensor and a magnet.

Hereinafter, the present invention will be described in more detail with reference to one or more embodiments. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Hereinafter, one or more embodiments will be described in more detail with reference to Examples. However, these examples are provided to illustrate one or more embodiments illustratively and the scope of the present invention is not limited to these examples.

Example 1 Induction of Binding of Influenza A Nucleoproteins and Particles

In order to induce influenza A nucleoprotein (hereinafter referred to as 'nucleoprotein') and particles to form a complex, the following experiment was carried out as follows. Is formed.

Specifically, 100 fM of magnetic particles of 0.5 μm and 10 fM of polystyrene beads of 2.8 μm are prepared, followed by EDC-NHS {(1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide) -(N-hydroxysuccinimide)} was activated by treatment. Thereafter, the activated particles and the nuclear protein antibody having different recognition sites were mixed with each other to induce binding of the particles with the antibodies. Next, each of the nucleoprotein (10 pM; experimental group) or bovine serum albumin (hereinafter referred to as 'BSA'; control group) dissolved in binding buffer (1X PBS and 0.01% Tween 20) was used as the antibody-. Contact with the particle complex for a sufficient time led to the formation of the protein-antibody-particle complex.

Example 2: Confirmation of Formation of Protein-Antibody-Particle Complex

In order to confirm the formation of the protein-antibody-particle complex tested in Example 1, a CMOS image sensor (hereinafter referred to as 'CIS') and a magnet were used.

First, the sensor surface image of the control group and the experimental group was photographed without contacting the magnet to the CIS, and the amount of polystyrene particles was measured through the image. The magnet was then brought into contact with the CIS, the sensor surface was cleaned, and the image was taken again. At this time, the polystyrene particles combined with the magnetic particles are not washed away, and the polystyrene particles not bonded with the magnetic particles are washed to an intensity that can be washed out. Thereafter, the ratio was calculated by comparing the amount of polystyrene particles before contacting the magnet with the amount of polystyrene particles remaining after washing.

As a result, as shown in FIG. 4, in the initial image before contacting the magnet, it was found that the magnetic particles and the polystyrene particles were mixed in both the experimental group and the control group. However, even after contacting the magnet and cleaning the sensor surface, it was confirmed that the polystyrene particles were bonded to the sensor surface, which means that the magnetic particle-nucleoprotein-polystyrene particle complex was also collected on the sensor surface.

In particular, when comparing the number of polystyrene particles before and after contacting the magnet, it was confirmed that the ratio of the collected polystyrene particles was significantly higher in the experimental group (Fig. 5). In the experimental group, since the nucleoprotein, polystyrene particles, and magnetic particles are bonded to each other to form a complex, it means that the magnetic particles are collected by the magnet and thus the polystyrene particles are also collected.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

100: target material detection unit
110: magnetic field generating unit
120: image sensor
130: thin film

Claims (7)

A magnetic field generating unit generating a magnetic field;
An image sensor connected in line with one side of the magnetic field generator;
A target material detection unit including a thin film attached to a surface of the image sensor; And
Includes two kinds of binding molecules that specifically bind to different recognition sites present in the target material,
Of the binding molecules, one type of binding molecule is a magnetic particle bound, the other type of binding molecule is a polystyrene particle bound,
The diameter of the polystyrene particles is 5 to 6 times larger than the diameter of the magnetic particles,
The two kinds of binding molecules specifically bind to the target material,
The target material is collected in the thin film by the magnetic field generating unit,
The collection is made without the passage of microfluidic channels,
The image sensor is a complementary metal oxide semiconductor image sensor,
The thin film is glass,
The target material detection kit that is made at the same time the imaging of the target material by the capture and the image sensor.
The method of claim 1,
The magnetic field generating unit is a kit for detecting a target material, characterized in that any one selected from the group consisting of a magnet, a solenoid and a superconducting magnet.
delete delete delete The method of claim 1,
The magnetic particles are a target material detection kit, characterized in that the size of 100 nm to 3 ㎛.
The method of claim 1,
The binding molecule kit for detecting a target substance, characterized in that selected from the group consisting of polypeptides, polynucleotides and lipids.

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