KR102071507B1 - Hydrogel structure for a biosensor, biosensor including the same, method of preparing the hydrogel structure for the biosensor - Google Patents

Abstract

A core comprising a first photocrosslinkable acrylic polymer; And a shell comprising a second photocrosslinkable acrylic polymer, a third photocrosslinkable acrylic polymer including a cationic functional group, and a microorganism detectable color change dye; a hydrogel structure for a biosensor comprising a core / shell structure. A biosensor comprising, and a method of manufacturing the hydrogel structure for the biosensor is disclosed.

Description

Hydrogel structure for a biosensor, a biosensor comprising the same, and a method of manufacturing the hydrogel structure for the biosensor {Hydrogel structure for a biosensor, biosensor including the same, method of preparing the hydrogel structure for the biosensor}

It relates to a hydrogel structure for a biosensor, a biosensor comprising the same, and a method for producing the hydrogel structure for the biosensor.

A biosensor is a device that analyzes a substance by an interaction represented by a biochemical receptor such as an enzyme, an antibody, an antigen, and the like to be measured.

Food biosensors are mainly used to check the stability of foods by detecting pathogens, residual pesticides, and other toxic chemicals.

In order to confirm such food stability, there is a need for a novel structure that is easy to carry and economically confirms the presence of food harmful bacteria in a short time.

One aspect is to provide a hydrogel structure for biosensors that is easy to carry and detects harmful foods in a short time and is economical.

Another aspect is to provide a biosensor comprising the hydrogel structure.

Another aspect is to provide a method of manufacturing a hydrogel structure for a biosensor capable of mass production at low cost.

According to one aspect,

A core comprising a first photocrosslinkable acrylic polymer; And

Provided is a hydrogel structure for a biosensor comprising a core / shell structure comprising a second photocrosslinkable acrylic polymer, a third photocrosslinkable acrylic polymer including a cationic functional group, and a microorganism detectable color change dye. .

According to another aspect,

There is provided a biosensor comprising the hydrogel structure described above.

According to another aspect,

Preparing a core solution including a first photocrosslinkable acrylic monomer, a first photoinitiator, a first photocrosslinker, and an organic solvent;

Preparing a shell aqueous solution including a second photocrosslinkable acrylic monomer, a third photocrosslinkable acrylic monomer including a cationic functional group, a second photoinitiator, a second photocrosslinker, and water; And

Contacting the core solution and the shell aqueous solution with an organic medium and performing a photopolymerization reaction to obtain a core / shell hydrogel structure; And

Preparing the hydrogel structure for the biosensor by impregnating the core / shell hydrogel structure into a microorganism detectable color change dye solution and adsorbing the microorganism detectable color change dye to the shell of the core / shell hydrogel structure; There is provided a method of manufacturing a hydrogel structure for a biosensor comprising a.

Hydrogel structure for a biosensor according to one aspect comprises a core / shell structure, the core / shell structure is a microorganism detectable color change dye is selectively located in the shell to stain the cell membrane of harmful bacteria by the diffusion of the dye. Can be. For this reason, it is possible to economically detect harmful bacteria only by changing the color of the shell.

In addition, the third photocrosslinkable acrylic polymer including a cationic functional group may be concentrated in the shell, thereby exhibiting high affinity through microorganisms that exhibit negative charges and electrostatic attraction. In addition, the production method of the hydrogel structure for the biosensor can be mass-produced at low cost.

1 is a schematic diagram showing a mechanism capable of detecting food corruption using a biosensor core / shell hydrogel structure (bead) according to one embodiment.
FIG. 2 is a schematic diagram showing improved affinity between the surface of the core / shell hydrogel structure with positive charge and the bacteria with negative charge.
3 is a schematic diagram illustrating a method of manufacturing a core / shell hydrogel structure (bead) using an electrospray apparatus according to one embodiment.
Figure 4 is a photograph of the cross-section of the beads after the impregnation of bacteria in the shell aqueous solution for 2 hours during the manufacturing process of the core / shell hydrogel beads of Example 1 with a camera.
FIG. 5 is a photograph of the core / shell hydrogel beads prepared according to Example 1 after immersing the bacterial solution in the bacterial solution for 6 hours.

With reference to the accompanying drawings, it will be described in detail a hydrogel structure for a biosensor, a biosensor comprising the same, and a method for producing the hydrogel structure for the biosensor according to an exemplary embodiment. The following is presented by way of example, and the present invention is not limited thereto. The present invention is defined only by the scope of the claims to be described later.

As used herein, the term " ~ acrylic polymer (monomer) " is used in a concept including " ~ acrylic polymer (monomer) ", and " derivatives of ~ acrylic polymer (monomer) ".

As used herein, the term "hydrogel structure" refers to a "structure in which water flows into a polymer network structure," and refers to a structure having high permeability and swelling including an aqueous solution therein.

As used herein, "core / shell structure" means a structure in which an interface is formed due to different heterogeneous phase separation therein, and defines a central side of the structure as a core from the interface and a outer side of the structure as a shell from the interface. .

In the food sector, the Rural Development Institute for Agricultural Engineering developed a technique for detecting Salmonella enterosis using impedance biosensors in 2006. This technology can detect up to 10,000 grains / ml in 3 minutes, but it is difficult to popularize because complex devices and analysis equipment are required to detect and analyze the substance.

As an alternative, color conversion sensors have been developed that detect colors that change due to interactions with the materials they contain when they detect a material to be measured. Since the color conversion sensor has a low sensitivity characteristic, a method of promoting redox reaction of gas using a redox compound such as ferrocene has been developed in order to increase the sensitivity characteristic. However, in the case of the redox compound itself is harmful, there is a limit to use as a sensor for food.

In order to solve this problem, the inventors of the present invention intend to propose a hydrogel structure for a biosensor as follows, a biosensor comprising the same, and a method of manufacturing the hydrogel structure for the biosensor.

Hydrogel structure for a biosensor according to an embodiment includes a core comprising a first photocrosslinkable acrylic polymer; And a shell including a second photocrosslinkable acrylic polymer, a third photocrosslinkable acrylic polymer including a cationic functional group, and a microorganism detectable color change dye.

The biosensor hydrogel structure may include a core / shell structure. In the hydrogel structure including the core / shell structure, the microbe detectable color change dye is located in the shell as compared to the general hydrogel structure, and the cell membranes of harmful bacteria may be dyed by diffusion of the dye. In addition, the third photocrosslinkable acrylic polymer including a cationic functional group may be concentrated in the shell, thereby exhibiting high affinity through microorganisms exhibiting a negative charge and electrostatic attraction. In addition, the biosensor hydrogel structure is easy to carry.

The core / shell structure may further include a water layer on its surface. On the surface of the positively charged shell including the water film layer, food harmful bacteria having negative charges such as Escherichia coli, E. coli, Salmonella, etc., which have negative charges, may be attached.

For example, the first photocrosslinkable acrylic polymer and the second photocrosslinkable acrylic polymer may include a polymer represented by Formula 1 below:

[Formula 1]

In Chemical Formula 1,

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ , R ₁₈ , R ₁₉ , R ₂₀ , R ₂₁ , R ₂₂ , R ₂₃ , R ₂₄ may be each independently a hydrogen atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof;

m, n, l, k may each be an integer of 5 to 100,000.

For example, in Formula 1, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ , R ₁₈ , R ₁₉ , R ₂₀ , R ₂₁ , R ₂₂ , R ₂₃ , R ₂₄ may each independently be a hydrogen atom or a substituted or unsubstituted C1 to C5 alkyl group. have.

The cationic functional group may comprise a quaternary ammonium radical cation. The quaternary ammonium radical cation causes a positive charge on the surface of the hydrogel structure. In addition, the quaternary ammonium radical cation may have different solubility in water depending on the number or polarity of the chains of the functional groups substituted in the ammonium salt.

For example, the third photocrosslinkable acrylic polymer including the cationic functional group may include a polymer represented by Formula 2 below:

[Formula 2]

In Chemical Formula 2,

R ' ₁ , R' ₂ , R ' ₃ , R' ₄ , R ' ₅ , R' ₆ , R ' ₇ , R' ₈ , R ' ₉ , R' ₁₀ , R _'11 , R' ₁₂ , R ' ₁₃ , R _'14 , R' ₁₅ , R _'16 , R' ₁₇ , R _'18 , R' ₁₉ , R _'20 , R' ₂₁ , R _'22 , R' ₂₃ , R _'24 are each independently hydrogen An atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof;

R ′ ₂₅ , R ′ ₂₆ , R ′ ₂₇ may be independently a substituted or unsubstituted C 1 to C 10 alkyl group, or a substituted or unsubstituted C 6 to C 20 aryl group, or a combination thereof;

p, q, r, and v may each be an integer of 5 to 100,000.

For example, in Formula 2, R _'25 , R' ₂₆ , R _'27 may be a substituted or unsubstituted C1 to C5 alkyl group independently of each other.

The microorganism detectable color change dye is MTT (3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide) or Rezazurin (7-Hydroxy-3 H -phenoxazin-3-one 10-oxide It can be a dye selected from). For example, the microorganism detectable color change dye may be MTT (3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide). However, the microorganism detectable color change dye is not limited thereto, and any microorganism detectable color change dye available in the art is possible.

The hydrogel structure may include beads, fibers, nonwovens, films, or a combination thereof. For example, the hydrogel structure may be beads or fibers. The beads may be spherical, elliptical, rod-shaped, or the like, for example, spherical.

The beads or fibers may have a diameter of 200 μm to 3 mm, for example a diameter of 500 μm to 3 mm, for example a diameter of 1 mm to 3 mm. The bead or fiber-type hydrogel structure may have a wider surface area than the film form, and thus may detect food harmful bacteria.

The core / shell structure may stain the cell membrane of the harmful bacteria by diffusion of a microorganism detectable color change dye located in the shell when the harmful bacteria adhere to the surface thereof, thereby changing the color of the shell to be distinguished from the color of the core.

The harmful bacterium is selected from Escherichia coli, E. coli, Salmonella, Staphylococcus aureus, enteritis Vibrio parahaemolyticus, Bacillus cereus, or a combination thereof. Can be.

1 is a schematic diagram showing a mechanism capable of detecting food corruption using a biosensor core / shell hydrogel structure (bead) according to one embodiment.

Referring to FIG. 1, a core / shell hydrogel structure (bead, 2) for a biosensor is a hydrogel structure composed of a core 5 and a shell 6. The core / shell hydrogel structure includes microorganism-detectable color change dye particles 1 in the shell 6, and a water layer 3 is disposed on the surface of the core / shell hydrogel structure. During food decay, for example, bacteria (4), which are food harmful bacteria, adhere to the surface of the water film layer (3), and the microorganism detectable color change dye particles (1), which are divided into a cationic portion and an anionic portion in an aqueous shell solution, By diffusion, the dye is combined with a substance that exhibits a negative charge of the plasma membrane inside the bacterial (4) cell membrane. At this time, the bacteria 4 are changed to the color changed bacteria 4 'and the color of the shell of the core / shell hydrogel structure (bead 2) for the biosensor is changed.

FIG. 2 is a schematic diagram showing improved affinity between the surface of the core / shell hydrogel structure with positive charge and the bacteria with negative charge.

Referring to FIG. 2, a chain portion 11 polymerized with a monomer of a second photocrosslinkable acrylic polymer in a shell of a core / shell hydrogel structure and a monomer of a third photocrosslinkable acrylic polymer having a cationic functional group (12) is connected with the optical crosslinking agent (13). During food decay, the bacteria 14 are electrostatically charged with the positively charged material and the negatively charged bacteria 14 of the chain portion 12 polymerized with a photocrosslinkable polymer monomer having a cationic functional group disposed in the shell. Affinity is enhanced by the attraction, so that the bacteria 14 can be easily attached and economically detect the attached bacteria 14.

The hydrogel structure may be included in the food storage package or food storage container surface if the patch or high permeability packaging material or food storage container. Therefore, the hydrogel structure can be miniaturized and portable.

Biosensor according to another embodiment may include the aforementioned hydrogel structure.

According to another embodiment, a method of manufacturing a hydrogel structure for a biosensor includes preparing a core solution including a first photocrosslinkable acrylic monomer, a first photoinitiator, a first photocrosslinker, and an organic solvent; Preparing a shell aqueous solution including a second photocrosslinkable acrylic monomer, a third photocrosslinkable acrylic monomer including a cationic functional group, a second photoinitiator, a second photocrosslinker, and water; Contacting the core solution and the shell aqueous solution with an organic medium and performing a photopolymerization reaction to obtain a core / shell hydrogel structure; And impregnating the core / shell hydrogel structure into a microorganism detectable color change dye solution and adsorbing the microorganism detectable color change dye to the shell of the core / shell hydrogel structure to prepare the hydrogel structure for the biosensor. It can include;

Contacting the organic medium may comprise spraying through a double nozzle of the electrospray device.

3 is a schematic diagram illustrating a method of manufacturing a core / shell hydrogel structure (bead) using an electrospray apparatus according to one embodiment. The electrospray device has a syringe 22 and a double nozzle 23 and is connected to a power source 21. The droplets are sprayed 24 through the double nozzle 23. The diameter of the hydrogel structure (bead) can be adjusted by the needle size of the nozzle, the distance between the collection and spinning nozzle tip, the solution injection rate, or the operating voltage. For example, the needle size of the nozzle may be adjusted in the range of 18G to 27G, the distance between the collecting portion and the spinning nozzle tip in the range of 1 cm to 10 cm, and the operating voltage in the range of 5 to 20 kV, respectively. The solution injection rate can be adjusted by varying the solution injection rate of the core and the shell within the range of 1 to 5 ml / h.

Method for producing a hydrogel structure for a biosensor according to an embodiment is as follows.

First, a core solution including a first photocrosslinkable acrylic monomer, a first photoinitiator, a first photocrosslinker, and an organic solvent may be prepared. Next, a shell aqueous solution including a second photocrosslinkable acrylic monomer, a third photocrosslinkable acrylic monomer including a cationic functional group, a second photoinitiator, a second photocrosslinker, and water may be prepared.

The first photocrosslinkable acrylic monomer and the second photocrosslinkable acrylic monomer may each include a monomer represented by Formula 3 below:

[Formula 3]

In Chemical Formula 3,

R ₂₅ or R ₂₆ may be independently of each other a hydrogen atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof.

The third photocrosslinkable acrylic monomer may include a monomer represented by Formula 4 below:

[Formula 4]

In Chemical Formula 4,

R _'30 , R' ₃₁ may be independently of each other a hydrogen atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof;

R ′ ₃₂ , R ′ ₃₃ and R ′ ₃₄ may be each independently a substituted or unsubstituted C1 to C10 alkyl group, or a substituted or unsubstituted C6 to C20 aryl group, or a combination thereof.

The first photoinitiator and the second photoinitiator independently of each other 2, 2-dimethoxy-2-phenylacetophenone, 2-methoxy-2-phenylacetophenone, 2-hydroxy-1- [4- (2- Hydroxyethoxy) phenyl] -2-methyl-1-propanone, acetophenone derivatives, camphorquinone and mixtures thereof.

The first photocrosslinker and the second photocrosslinker are independently of each other ethylene glycol dimethacrylate, 1,3-butanediol dimethacrylate, 1,3-butanediol diacrylate, 1,4-butanediol dimethacrylate , 1, 4-butanediol diacrylate, 1, 6-hexanediol diacrylate, pentaerythritol triacrylate, tetraethylene glycol dimethacrylate, divinyl benzene, trimethylolpropane triacrylate, isophorone diisocyanate It may be at least one selected from glycidyl methacrylate, trimethylolpropane trimethacrylate, or a mixture thereof.

The organic solvent may be at least one selected from benzene, toluene, styrene, xylene, ethanol and ether.

Next, the core solution and the shell aqueous solution are contacted with an organic medium and subjected to photopolymerization to obtain a core / shell hydrogel structure.

In detail, the core / shell hydrogel structure inserts the core solution and the shell aqueous solution into the syringe 22, injects the same into the double nozzle 23, and drops the sprayed droplet 24 onto the organic medium. The spray droplets 24 are phase-separated due to the formation of an interface between the oil phase and the water, and the photopolymerization using a UV source therein as the spray droplets 24 are separated from the organic medium. The reaction is carried out to obtain a core / shell hydrogel structure (bead, 26). The photopolymerization reaction using the UV source may include radicals of a first photoinitiator or a second photoinitiator due to the UV source, and include a first photocrosslinkable acrylic monomer or a second photocrosslinkable acrylic monomer including a first photocrosslinker or a second photocrosslinker. Crosslinking is performed between carbon double bonds (= CH ₂ ) of the third photocrosslinkable acrylic monomer.

Next, the core / shell hydrogel structure (bead, 26) is impregnated with a microorganism detectable color change dye solution, and the microorganism detectable color change dye is adsorbed on the shell of the core / shell hydrogel structure (bead, 26). To prepare a hydrogel structure for a biosensor. The production method of the hydrogel structure for the biosensor is possible to mass production by reducing the cost.

The organic medium may comprise mineral oil. The organic medium may be used in addition to the mineral oil, an organic medium usable in the art.

Looking at the definition of the substitution (group) used in the formula of the present specification are as follows.

The "substituted" having an alkyl group or an aryl group includes a halogen atom, a C1 to C10 alkyl group substituted with a halogen atom (eg, CCF ₃ , CHCF ₂ , CH ₂ F, CCl _3, etc.), a hydroxy group, a nitro group, a cyano group, an amino group, Amidino groups, hydrazines, hydrazones, carboxyl groups or salts thereof, sulfonic acid groups or salts thereof, phosphoric acid or salts thereof, alkyl groups of C1 to C10, alkenyl groups of C2 to C10, alkynyl groups of C2 to C10, heteros of C1 to C20 It means substituted with an alkyl group, a C6 to C20 aryl group, a C6 to C20 arylalkyl group, a C6 to C20 heteroaryl group, or a C6 to C20 heteroarylalkyl group.

Specific examples of the C1 to C10 alkyl group used in the formula include methyl, ethyl, propyl, isobutyl, sec-butyl, ter-butyl, neo-butyl, iso-amyl, or hexyl, and the like. The above hydrogen atoms may be substituted with substituents as defined in the above-mentioned "substitution".

Specific examples of the C6 to C20 aryl group used in the formula include phenyl, naphthyl, tetrahydronaphthyl and the like, and at least one hydrogen atom of the aryl group is substituted with a substituent as defined in the aforementioned "substitution". It is possible.

Specific examples of the C2-C10 alkenyl group used in the "substitution" include vinylene, allylene and the like.

Specific examples of the C2-C10 alkynyl group used in the "substitution" include acetylene and the like.

C1 to C20 heteroalkyl group or arylalkyl group used in the "substituted" means that one or more of the carbon atoms constituting the alkyl group is replaced with a hetero atom such as N, O, S, or P or an aryl group such as phenyl .

The aryl group of C6 to C20 used in the "substitution" is used alone or in combination, and means an aromatic system containing one or more rings, for example phenyl, naphthyl, tetrahydronaphthyl and the like. .

The C6 to C20 heteroaryl group used in the "substituted" means one or more heteroatoms selected from N, O, P or S, and means that the remaining ring atom is an organic compound of carbon, for example pyridyl and the like Can be mentioned.

C6 to C20 heteroarylalkyl group used in the above "substituted" means that one or more of the carbon atoms constituting the alkyl group is replaced with the aforementioned heteroaryl group.

Hereinafter, examples of the present invention will be described. However, the following examples are only examples of the present invention and the present invention is not limited to the following examples.

EXAMPLE

Example  1: core / shell Hydrogel Bead  Produce

A first photocrosslinkable acrylic monomer (HEMA) represented by the following formula (3-1), ethylene glycol dimethacrylate (EGDMA) as the first photocrosslinker, 2,2-dimethoxy-2-phenylacetophenone (first photoinitiator) DMPA), and toluene were prepared in a core solution in a molar ratio of 100: 1: 1: 1 respectively.

A second photocrosslinkable acrylic monomer (HEMA) represented by the following formula (3-1) and a third photocrosslinkable acrylic monomer (MAETC) represented by the following formula (4-1), wherein the concentration of MAETC is fixed at 50 mol% relative to the HEMA ), Ethylene glycol dimethacrylate (EGDMA) as the second photocrosslinker, 2,2-dimethoxy-2-phenylacetophenone (DMPA) as the second photoinitiator, and water, respectively, in a molar ratio of 100: 1: 1: 1. Aqueous shell solution was prepared.

The core solution and the shell aqueous solution were supplied with a transformer (230 High Voltage Power Supply, manufactured by Spellman) and a syringe pump (KDS100, manufactured by KD Scientific, Inc.) (distance between collector and spinning nozzle tip: 5 cm Injection into the dual nozzles (inner diameter: 400 μm, outer diameter: 900 μm) (core injection rate: 1 ml / h, shell injection rate: 2 ml / h) Sprayed to oil, performing a photopolymerization reaction using a UV source (wavelength: 365 nm) to a core comprising a first photocrosslinkable acrylic polymer represented by the formula (1-1), and a second represented by the formula (1-1) A core / shell hydrogel bead consisting of a shell comprising a photocrosslinkable acrylic polymer and a third photocrosslinkable acrylic polymer including a cationic functional group represented by 2-1 below was prepared.

[Formula 1-1]

In Chemical Formula 1-1,

a, b, c and d are each about 100 to 1,000.

[Formula 2-1]

In Chemical Formula 2-1,

a ₁ , b ₁ , c ₁ , and d ₁ are each about 100 to 1,000.

 [Formula 3-1]

[Formula 4-1]

Analysis example  1: Check core / shell structure

During the preparation of the core / shell hydrogel beads of Example 1, the aqueous solution of the shell was impregnated with bacteria for 2 hours, and the cross section of the beads was observed using a camera (VH-310G2, manufactured by Nikon). The results are shown in FIG.

As bacteria, Escherichia coli (E. coli) produced during food decay was used. The solution containing Escherichia coli (E. coli) contained in the shell fixed the OD value to 1 at 600 nm wavelength, and the concentration of the solution containing Escherichia coli (E. coli) was about 0.6 at an OD value of 1. ˜2 × 10 ⁹ (cells / mL). Whether bacteria were attached to the surface of the hydrogel beads was confirmed by an assay using an MTT solution. The MTT solution used was a solution in which triazole blue tetrazolium bromide powder was dissolved in phosphage buffer saline (PBS) suspension at a concentration of 5 mg / mL.

Referring to FIG. 4, the color of the core of the hydrogel beads prepared in Example 1 was not changed from yellow, but the color of the shell was changed from yellow to purple.

From this, it can be confirmed that the core / shell hydrogel bead of Example 1 is attached to the shell of the hydrogel bead when Escherichia coli (Escherichia coli, E. coli) is a harmful bacterium. This is believed to be due to the formation of formazan crystals by reduction of tetrazolium salts by dehydrogenase in the mitochondria of Escherichia coli cells due to the metabolism of Escherichia coli (E. coli), a harmful bacterium.

The color of the shell of the core / shell hydrogel beads of Example 1 is distinguished from the color of the core, thereby confirming that the hydrogel beads have a core / shell structure.

Evaluation example  1: Evaluation of harmful bacteria detection

The core / shell hydrogel beads prepared in Example 1 were impregnated with the bacterial solution for 6 hours and then observed with a camera (VH-310G2, manufactured by Nikon). The results are shown in FIG.

As bacteria, Escherichia coli (E. coli) produced during food decay was used. The bacterial solution was fixed at an OD value of 1 at 600 nm wavelength, and the concentration of E. coli (Escherichia coli) solution was about 0.6-2 × 10 ⁹ (cells / mL) when the OD value was 1. Whether bacteria were attached to the surface of the hydrogel beads was confirmed by an assay using an MTT solution. The MTT solution used was a solution in which triazole blue tetrazolium bromide powder was dissolved in phosphage buffer saline (PBS) suspension at a concentration of 5 mg / mL.

Referring to FIG. 5, the color of the core / shell hydrogel beads prepared by Example 1 changed from yellow to purple. From this, it can be confirmed that Escherichia coli (Escherichia coli, E. coli), which is a harmful bacterium, is attached to the surface of the core / shell hydrogel bead during food decay. This is believed to be due to the formation of formazan crystals by reduction of tetrazolium salts by dehydrogenase in the mitochondria of Escherichia coli cells due to the metabolism of Escherichia coli (E. coli), a harmful bacterium.

1: microbe detectable color change dye particles,
2: core / shell hydrogel structure (bead) for biosensor,
3: water layer, 4: bacteria, 4 ': color changed bacteria,
5, 15: core, 6, 16: shell, 11: chain portion polymerized with the monomer of the second photocrosslinkable acrylic polymer, 12: chain portion polymerized with the monomer of the third photocrosslinkable acrylic polymer having a cationic functional group,
13: photocrosslinker, 14: bacteria, 21: power source,
22: syringe, 23: double nozzle, 24: spray droplets,
25: organic medium, 26: core / shell hydrogel structure (bead)

Claims (23)

A core comprising a first photocrosslinkable acrylic polymer; And
And a core / shell structure consisting of a second photocrosslinkable acrylic polymer, a third photocrosslinkable acrylic polymer including a cationic functional group, and a microbial detectable color change dye;
The core / shell structure is a biosensor, in which the cell membrane of the harmful bacterium is stained by the diffusion of the microorganism detectable color change dye located in the shell when the harmful bacterium is attached to the surface, and the color of the shell is changed to distinguish the color of the core. Hydrogel structure for. The method of claim 1,
The core / shell structure is a hydrogel structure for a biosensor further comprising a water layer (water layer) on the surface. The method of claim 1,
The first photocrosslinkable acrylic polymer and the second photocrosslinkable acrylic polymer are hydrogel structures for biosensors comprising a polymer represented by Formula 1 below:
[Formula 1]

In Chemical Formula 1,
R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ , R ₁₈ , R ₁₉ , R ₂₀ , R ₂₁ , R ₂₂ , R ₂₃ , R ₂₄ are independently of each other a hydrogen atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof;
m, n, l, k are integers of 5-100,000, respectively. The method of claim 3,
In Formula 1, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ , R ₁₈ , R ₁₉ , R ₂₀ , R ₂₁ , R ₂₂ , R ₂₃ , R ₂₄ are each independently a hydrogen atom or a substituted or unsubstituted C1 to C5 alkyl group. Structure. The method of claim 1,
The cationic functional group is a hydrogel structure for a biosensor comprising a quaternary ammonium radical cation. The method of claim 1,
The third photocrosslinkable acrylic polymer including the cationic functional group is a hydrogel structure for a biosensor comprising a polymer represented by the following formula (2):
[Formula 2]

In Chemical Formula 2,
R ' ₁ , R' ₂ , R ' ₃ , R' ₄ , R ' ₅ , R' ₆ , R ' ₇ , R' ₈ , R ' ₉ , R' ₁₀ , R _'11 , R' ₁₂ , R ' ₁₃ , R _'14 , R' ₁₅ , R _'16 , R' ₁₇ , R _'18 , R' ₁₉ , R _'20 , R' ₂₁ , R _'22 , R' ₂₃ , R _'24 are each independently hydrogen An atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof;
R ′ ₂₅ , R ′ ₂₆ , R ′ ₂₇ are each independently a substituted or unsubstituted C 1 to C 10 alkyl group, or a substituted or unsubstituted C 6 to C 20 aryl group, or a combination thereof;
p, q, r, and v are each an integer of 5 to 100,000. The method of claim 6,
In Formula 2, R _'25 , R' ₂₆ , R _'27 are independently substituted or unsubstituted C1 to C5 alkyl group of the biosensor hydrogel structure. The method of claim 1,
The microorganism detectable color change dye is MTT (3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide) or Rezazurin (7-Hydroxy-3 H -phenoxazin-3-one 10-oxide Hydrogel structure for biosensor which is at least one dye selected from). The method of claim 1,
The hydrogel structure is a hydrogel structure for a biosensor comprising a bead (bead). The method of claim 9,
The bead or fiber is a hydrogel structure for a biosensor having a diameter of 200 ㎛ to 3 mm. delete The method of claim 1,
The harmful bacterium is selected from Escherichia coli, E. coli, Salmonella, Staphylococcus aureus, Vibrio parahaemolyticus, Bacillus cereus, or a combination thereof. Hydrogel structure for phosphorus biosensor. A biosensor comprising a hydrogel structure according to any one of claims 1 to 10 and 12. Preparing a core solution including a first photocrosslinkable acrylic monomer, a first photoinitiator, a first photocrosslinker, and an organic solvent;
Preparing a shell aqueous solution including a second photocrosslinkable acrylic monomer, a third photocrosslinkable acrylic monomer including a cationic functional group, a second photoinitiator, a second photocrosslinker, and water;
Contacting the core solution and the shell aqueous solution with an organic medium and performing a photopolymerization reaction to obtain a core / shell hydrogel structure; And
The method of claim 1, wherein the core / shell hydrogel structure is impregnated with a microorganism detectable color change dye solution and the microorganism detectable color change dye is adsorbed to the shell of the core / shell hydrogel structure. A method for producing a hydrogel structure for a biosensor, comprising the steps of: preparing a hydrogel structure for a biosensor according to claim 1. The method of claim 14,
Contacting the organic medium comprises spraying through a double nozzle of an electrospray device. The method of claim 14,
The first photocrosslinkable acrylic monomer and the second photocrosslinkable acrylic monomer is a method for producing a hydrogel structure for a biosensor, each comprising a monomer represented by the following formula (3):
[Formula 3]

In Chemical Formula 3,
R ₂₅ or R ₂₆ are independently of each other a hydrogen atom, a substituted or unsubstituted C1 to C10 alkyl group, or a combination thereof. The method of claim 14,
Method for producing a hydrogel structure for a biosensor comprising the monomer represented by the third photocrosslinkable acrylic monomer comprising the cationic functional group:
[Formula 4]

In Chemical Formula 4,
R ′ ₃₀ , R ′ ₃₁ are independently of each other a hydrogen atom, a substituted or unsubstituted C 1 to C 10 alkyl group, or a combination thereof;
R _'32 , R' ₃₃ and R _'34 are each independently a substituted or unsubstituted C1 to C10 alkyl group, or a substituted or unsubstituted C6 to C20 aryl group, or a combination thereof. The method of claim 14,
The third photocrosslinkable acrylic monomer containing the cationic functional group is a method of producing a hydrogel structure for a biosensor for which the mole fraction is 10 mol% to 70 mol% with respect to the second photocrosslinkable acrylic monomer in a shell aqueous solution. The method of claim 14,
The first photoinitiator and the second photoinitiator independently of each other 2, 2-dimethoxy-2-phenylacetophenone, 2-methoxy-2-phenylacetophenone, 2-hydroxy-1- [4- (2- Hydroxyethoxy) phenyl] -2-methyl-1-propanone, acetophenone derivative, camphorquinone and a method for producing a hydrogel structure for a biosensor which is at least one selected from these mixtures. The method of claim 14,
The first photocrosslinker and the second photocrosslinker are independently of each other ethylene glycol dimethacrylate, 1,3-butanediol dimethacrylate, 1,3-butanediol diacrylate, 1,4-butanediol dimethacrylate , 1, 4-butanediol diacrylate, 1, 6-hexanediol diacrylate, pentaerythritol triacrylate, tetraethylene glycol dimethacrylate, divinyl benzene, trimethylolpropane triacrylate, isophorone diisocyanate A method for producing a hydrogel structure for a biosensor which is at least one selected from glycidyl methacrylate, trimethylolpropane trimethacrylate, or mixtures thereof. The method of claim 14,
The organic solvent is at least one selected from benzene, toluene, styrene, xylene, ethanol, ether. The method of claim 14,
Bio-sensor for biosensor further comprising a step of producing a core / shell structure by performing phase polymerization and photopolymerization reaction due to the interface between the oil phase and the water after contacting the organic medium Method for producing a gel structure. The method of claim 14,
The organic medium is a method for producing a hydrogel structure for a biosensor comprising a mineral oil.

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