KR20100112662A - Polyelectrolyte multilayer and biosensor - Google Patents

Abstract

PURPOSE: A polyeletrolyte multilayer and biosensor using the same are provided to ensure high cell adsorption and wetting property and to measure specific material content. CONSTITUTION: A polyelectrolyte multilayer comprises a cyclodextrin or derivative thereof in a laminated structure of an anionic polymer electrolyte. The anionic polyelectrolyte multilayer contains polymethacrylic acid, polyacrylic acid or hyaluronic acid. The cyclodextin is alpha-cyclodextrin or beta-cyclodextrin. A biosensor contains polyelectrolytic multimer.

Description

POLYELECTROLYTE MULTILAYER AND BIOSENSOR}

The present invention relates to a polymer electrolyte multilayer membrane and a biosensing device.

Recently, bioengineering has emerged. Bioengineering refers to the characteristics of cells through interaction between the support and the cells where cells are attached and grow, unlike conventional methods of controlling hormones, growth factors, and serum contained in cell culture in studying or inducing cell functions. It refers to the regulation of phosphorus attachment, proliferation, differentiation and extracellular matrix secretion. This requires the development of materials with biocompatibility and the development of chemical surface modification.

Therefore, a variety of multilayer membranes including a multilayer membrane having not only biocompatibility but also improving cell adhesion, and a multilayer membrane having both capturing ability and cell adhesion ability to a specific material and biosensors including the same Development is required.

In order to meet the above demands, an object of the present invention is to provide a polymer electrolyte multilayer film having excellent biocompatibility and having a selective capturing ability to a material, and a biosensing device including the multilayer film.

In order to achieve the above object, the present invention provides a multilayer membrane including cyclodextrin or a derivative thereof in an alternating stacked structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer.

In order to achieve the above object, the present invention includes an alternating layered structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer, wherein one or both layers of the outermost layer of the alternating layered structure is made of a cyclodextrin or a derivative thereof A multilayer film is provided.

In addition, the present invention provides a biosensing device comprising the multi-layer film in order to achieve the above object.

The inventors of the present invention prepare a multilayer film using a layer-by-layer (LBL) deposition technique using a polymer electrolyte that is safest in terms of immunity and toxicity, and can secure antimicrobial and antibacterial properties, and the cyclodextrin or When including or adsorbing derivatives thereof, or further laminating a film formed of a polymer made of a cyclodextrin or a derivative thereof on the outermost layer of the multilayer membrane, it is suitable for cell growth and hygroscopicity, and a specific material is collected according to application conditions or The present invention has been accomplished by confirming that it can induce optical activity of certain materials.

Hereinafter, the present invention will be described in more detail.

In the present invention, the polymer electrolyte refers to a polymer capable of dissociating in a solvent, and the polymer electrolyte may include a cationic polymer electrolyte and an anionic polymer electrolyte.

The cationic polymer electrolyte may mean a polymer electrolyte in which the chemical functional group of the polymer chain loses an anion or obtains hydrogen ions in an aqueous solution, and thus may be positively charged. The degree of ionization may be affected by the acidity or basicity of the surrounding environment. In addition to the various weakly electrolytic cationic polymers, neutral water-soluble polymers include neutral water-soluble polymers such as polyacrylamide, which are not ionized well in the general pH range of water and have a positive charge when the acidity of the surroundings is very high.

The anionic polymer electrolyte may mean a polymer electrolyte in which a polymer functional group of the polymer chain loses a cation or loses hydrogen ions, and thus may be negatively charged. The degree of ionization may vary depending on the acidity or basicity of the surrounding environment. Weak electrolyte anionic polymers are included.

In the present invention, 1 bilayer means a multilayer film in which one layer of each of two polymer electrolyte layers having different kinds is stacked.

In one aspect of the invention, the invention relates to a multilayer film.

The multilayer film may be a polymer electrolyte multilayer film, and more specifically, a multilayer film including an alternating stack structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer or the alternating stack structure, and the outermost layer of the alternating stack structure. Either layer or both layers may be a multilayer film made of a polymer of cyclodextrin or a derivative thereof.

The anionic polymer electrolyte layer is preferably poly methacrylic acid (PMA), poly acrylacid (PAA), polystyrene sulfonate (PSS) and hyaluronic acid (HA) It may be made of any one selected from the group consisting of), the cationic polymer electrolyte layer is preferably poly allylamine hydrochloride (poly (allylamine hydrochloride) (PAH), poly acrylamide (PAAm) and poly 4 It may be made of any one selected from the group consisting of -vinylbenzyltrimethylammonium chloride (poly (4-vinylbenzlytri-methyl ammonium chloride, PVTAC).

The multilayer membrane may be a multilayer membrane having at least one cationic polymer electrolyte layer and one anionic polymer electrolyte layer. For example, the multilayer film may be 1 bilayer or more, and may be two or more layers, respectively.

In the present invention, the layer-by-layer (LBL) refers to a method of controlling the thickness of the polymer electrolyte multilayer film or the film by repeating as many times as desired so that the polymer electrolyte having a relative charge is adsorbed on the substrate surface. .

In addition, when the cationic polymer electrolyte layer or the anionic polymer electrolyte layer is two or more, the electrolyte layer may be laminated with the same polymer electrolyte layer or different polymer electrolyte layers, respectively. Specifically, when the anionic polymer electrolyte layer is two or more layers, specific anionic polymer electrolyte layers may be alternately stacked with the cationic polymer electrolyte layer, or different types of anionic polymer electrolyte layers may be alternately stacked with the cationic polymer electrolyte layer. Can be. The cationic polymer electrolyte layer may be the same as above.

The outermost layer refers to the outermost layer of the multilayer film, that is, the outermost layer, that is, the polymer electrolyte layer located at the outermost layer.

The polymethacrylic acid may include not only polymethacrylic acid itself but also salts thereof. Specifically, at least one selected from the group consisting of the polymethacrylic acid and polymethacrylic acid salts. The molecular weight of the polymethacrylic acid is not particularly limited, but may be, for example, 10,000 to 200,000 or 50,000 to 150,000.

The polyacrylic acid may include not only polyacrylic acid itself but also a salt thereof. Specifically, it may be at least one selected from the group consisting of the polyacrylic acid and polyacrylic acid salt. The molecular weight of the polyacrylic acid is not particularly limited, but may be, for example, 10,000 to 100,000 or 30,000 to 90,000.

The hyaluronic acid is a kind of glucosamide glycan consisting of a disaccharide unit in which D-glucuronic acid and N-acetylglucosamine are connected by β (1 → 3) glycosidic bonds, and the hyaluronic acid has a chemical and physical structure. There are no species differences, humans have a metabolic system and are the safest biomaterials in terms of immunity and toxicity.

The hyaluronic acid may include not only hyaluronic acid itself but also all salts thereof. Specifically, the hyaluronic acid may be one or more selected from the group consisting of hyaluronic acid and hyaluronic acid salts. The hyaluronic acid salt includes both inorganic salts such as sodium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronic acid and the like and organic salts such as tetrabutylammonium hyaluronic acid. The molecular weight of the hyaluronic acid is not particularly limited, but may be, for example, 100,000 to 10,000,000.

The polyallylamine hydrochloride may be chemically bonded with hyaluronic acid or polyacrylic acid, and the molecular weight of the polyallylamine hydrochloride is not particularly limited, but may be, for example, 10,000 to 100,000 or 30,000 to 80,000.

The poly acrylamide may be chemically bonded with hyaluronic acid or polyacrylic acid, and the molecular weight of the poly acrylamide is not particularly limited, but may be, for example, 100,000 to 10,000,000.

When the cationic polyelectrolyte is polyacrylamide, the multi-layered film may have cross-linking formed by performing a crosslinking reaction between the polymer electrolytes in order to impart stability to the biological environment. The crosslinking reaction may be performed by, for example, a chemical reaction or heat treatment through EDAC.

The multilayer film of the present invention has a cyclodextrin or a derivative thereof in the multilayer film including the alternating stack structure of the polymer electrolyte layer or includes the alternating stack structure, and any one or both layers of the outermost layer of the alternating stack structure It may be a multilayer film made of a polymer of cyclodextrin or a derivative thereof.

The cyclodextrin is a cyclic oligosaccharide in which glucopyranose units bind α- (1,4), and typically 6 units of alpha-cyclodextrin (α-cyclodextrin). Dextrin), 7 units of beta-cyclodextrin (β-cyclodextrin), 8 units of gamma-cyclodextrin (γ-cyclodextrin), and the like. In addition, the cyclodextrin derivative may be a hydroxyl group (hydroxyl) bonded to each carbon may be substituted with another functional group, for example, carbon 2, 3 and 6 of the glucopyranose unit as shown in the following formula (1) The at least one hydroxyl group selected from the group consisting of hydroxyl groups bonded to may be substituted with a functional group or another functional group including an amine group.

[Formula 1]

The multilayer film may be applied to a device or a device requiring various polymer multilayer films including a biosensing device.

In one embodiment, the present invention provides a biosensing device including the multilayer film.

In the case of the multilayer membrane, the polymer electrolyte or the cyclodextrin polymer may be patterned for purposes such as selective adsorption of cells or for application of a biosensing device.

A biomaterial fixing material may be combined with the multilayer pattern layer formed through the patterning. For example, the biomaterial may be any one selected from the group consisting of cells, monosaccharides, disaccharides, oligosaccharides, fatty acids, polypeptides, proteins, and polynucleotides, and preferably cells. The cell may be any cell including prokaryotic and eukaryotic cells, for example fibroblasts, hepatocytes, neurons, cancer cells (eg HeLa cells), B cells, leukocyte cells ( white blood cells such as Raw264.7), and the like, including immune cells and embryonic cells.

The biomaterial-fixing material may be a biomaterial-fixing biomaterial. The biomaterial for fixing the biomaterial may be any oligopeptide including an oligopeptide sequence having an amino acid sequence of RGD and a protein including the oligopeptide and other receptor peptides, and the protein including the oligopeptide may be fibronectin ( fibronectin), fibrin, or the like, and the protein having the receptor peptide may be laminin, collagen, or the like.

In addition, the multilayer film may further include a thin film on the pattern layer, the thin film may include the biomaterial, preferably cells, the biomaterial is the biomaterial for fixing the biomaterial, Preferably, the polypeptide may include an oligopeptide sequence having an amino acid sequence of RGD. The thin film may be attached to a pattern layer including a polymer of a polymer electrolyte or a cyclodextrin or a derivative thereof. Preferably, the cells included in the thin film are a polymer electrolyte or a cyclodextrin or a derivative thereof in which a cell biomaterial is bound. It may be attached to a pattern layer containing a polymer of.

When the multilayer film is used in a DNA chip, protein chip, or cell-based biosensor, the outermost layer not including the pattern layer may be a substrate adhesive layer bonded to a substrate. In addition, when the multilayer film is used as a surface facing the damaged tissue for tissue regeneration inducing function, such as used for induced tissue regeneration, or as a dressing material for skin or mucosal tissue, the outermost layer not including the pattern layer May be a tissue adhesion layer that bonds to living tissue.

In another aspect of the invention, the invention relates to a bioreactor.

The bioreactor may be a biosensing device including the multilayer membrane.

Specifically, the bioreactor is

Board; The outermost layer may be a biosensing device including the multi-layered film formed on the substrate such that a polymer electrolyte layer or a layer made of a polymer of the cyclodextrin or a derivative thereof is a top layer.

The biosensing device is combined with a positively charged polymer electrolyte containing an amide group in a cell-fixing biomaterial, and thus can be applied to a cell-based biosensing device because it has excellent chemical stability and can provide conditions suitable for cell culture. . The biosensing device may be, for example, a biosensor.

The substrate is a substrate that can be applied to the biosensing device, the material is not particularly limited, for example, made of glass (glass slide) or metal substrate such as silicon (silicon) or gold (gold) It may have been. Specifically, the substrate may be a substrate that tends to be positively charged, such as mica, and may be a substrate that tends to be negatively charged, such as metal substrates such as glass, silicon, gold, and steel. It may be an organic substrate having no charge such as plastics (polystyrene, acrylic, Teflon, etc.).

In the case of the charged substrate, the multilayer film may be coated using a polymer electrolyte of opposite charge, and in the case of the organic substrate in the absence of the charge, the multilayer film may be coated using hydrophobic adsorption between polymer materials. .

The multilayer membrane may be one in which an anionic polymer electrolyte layer and a cationic polymer electrolyte layer are manufactured by using a layer-by-layer (LBL) deposition technique, and include a cyclodextrin or a derivative thereof, or a polymer of a cyclodextrin or a derivative thereof. It may be the outermost layer.

Since the biosensing device of the present invention and its manufacturing method use a polymer, it is possible to improve the adsorption of cells, to adsorb a specific material inside the multilayer film, and to improve the optical activity of the specific material. Not only can be used for various purposes including, but also combined with biological tissues can be used for experiments or diagnostics, medical or disease treatment, the industrial effect will be very large.

Hereinafter, preferred embodiments of the present invention will be described. However, the following examples are only examples for illustrating the present invention, and thus the protection scope of the present invention is not limited to the following examples.

Example :

Example  1-1. reagent

Poly (methacrylic acid) (PMA, Mw = 100,000) and Poly (allyamine hydrochloride) (PAH, Mw = 60,000) were used as reagents of Polysciences, and Carboxlyated beta-cyclodextrin was used as reagents of SUPELCO. Methylene blue Hydtrate was used by Fluka. All polymer electrolytes and reagents were used without further purification. Dulbecco's Phosphate Buffered Saline (DPBS), 0.25% Tripsinf-EDTA, and Dulbecco's modified eagle medium (DMEM) were used for cell experiments. In addition, Gibco fetal bovine serum (FBS) and Antibioic-Antimycotic were used to make DMEM.

Alpha-Cyclodextrin reagent was used by TCI, and beta-Cyclodextrin reagent was used by Aldrich. Epichlorohydrin (EP) and choline chloride (CC) were used from Aldrich. Dialysis was performed using Spectrum's molecular weight cut-off 1000 membrane. The whole process of dialysis and experiment was made with deionized water (> 18MΩcm Millipore Milli-Q).

Example  1-2: polymer Multilayer  Produce

The process of making and washing the polyelectrolyte aqueous solution was performed using deionized water (> 18 MΩ) produced using Millipore's Milli-Q water purifier.

PAH and PMA aqueous solution of the polymer electrolyte was made to a concentration of 0.01M and filtered through a filter having a pore of 0.45㎛ used to remove dust and microorganisms, and pH was adjusted with 0.1M HCl or NaOH aqueous solution. In the present invention, the polymer electrolyte multilayer membrane was formed by ionic bonding using positive and negative charges of the polymer electrolyte to form a polymer electrolyte multilayer membrane, and PAH and PMA were alternately adsorbed. In this experiment, the substrate (substrate) used for the measurement of the properties of the multilayered film was placed with a detergent diluted with silicon glass (silicon glass or silicon wafer) in a washing bottle, followed by ultrasonication for 15 minutes, and then rinsed again with deionized water. give. After 2-3 rinsing cycles, dry with nitrogen gas. After soaking in PAH cationic aqueous solution for 20 minutes, and then rinsing twice, immersing the substrate in PAA solution for 15 minutes and rinsing twice with deionized water for 2 minutes. Call this process 1 bilayer and increase the number of bilayers to the desired thickness. After the film is finished, it is quickly dried with nitrogen gas.

Example  1-3: multilayer thin film Cyclodextrin Including (adsorption)  Experiment

The experiment of including (adsorbing) the cyclodextrin was performed by the method shown in FIG. 1.

More specifically, the film on which the multilayer thin film of Example 1-2 was formed was immersed in Carboxylate beta cyclodextrin for 15 minutes.

Example 2: Checking the properties of the polymer multilayer

Example 2-1: Measurement of Wetting through Measurement of Contact Angle

In order to measure the wettability (wetability, wettability) of the multilayer film prepared in Example 1-3, the contact angle of water was measured using a CA-A Contact angle analyzer of Face. Advancing contact angle and receding contact angle, which are the contact angles identified by the contact angle measurement, are indices that help to measure the change tendency of the film with respect to moisture. In general, the wettable film is not only low in static contact angle but also reported in low advancing contact angle and receding contact angle.

The water used for measuring the contact angle was used after filtering the purified water, using a syringe to form a drop of 10 ul and then slowly raise the measuring plate on which the film is placed so as to contact the water drop so that the influence of gravity on the contact angle measurement It was. More specifically, as shown in FIG. 2, the static contact angle is measured, and the advancing contact angle is measured by further adding 10 ul of water in the state of measuring the static contact angle, and measuring the contact angle again. The receding contact angle was measured by sucking 10 ul of water from 20 ul of water droplets for measuring the advancing contact angle and measuring the contact angle again. The results are shown in FIG. 2.

As shown in FIG. 2, in the case of the polymer electrolyte multilayer membrane prepared in the present invention, when the beta-cyclodextrin is included, the contact interval is reduced by about half, and it is confirmed that the wettability is remarkably improved when the thickness of the membrane is different. .

Example  2-2: UV  How to measure

In the UV-vis spectrophotometer, when the ultraviolet light source passes through the sample, an electron transition occurs inside the sample. The frequency and intensity appear differently. Here, the absorbance of the material is measured according to the wavelength (nm) or the frequency (Hz) of the light and the data is displayed. It can also be measured in solution and used to see how well the film is formed. To explain how to use the Scinco S-3100 model used in the laboratory, first turn on the UV-vis spectrophotometer 20 minutes before using the program, and then run the program after 20 minutes. After executing the program, set up the basics (y-axis, number of repetitive measurements, etc.) and grab the base line before taking the sample. The base line is used as the slide glass cleaned by ultrasonication as well as the substrate secretion of the sample. For the base line, click the run blank (or the gray beacon in the toolbox) on the Measure menu to measure the base line after 10 scans. The sample is then measured. The sample is placed in the holder and can be measured by clicking run sample on the measure menu. Data is immediately graphed on the screen. To save the measured data, select File-Save-sample spectrum, CSV format file and press ok. Data is obtained in the form of a table. If you save the saved file in Excel and chart it, you can get the data in graph form.

2- 3atoms  Force microscope AFM Surface Morphological Characteristics Using

In order to examine the properties of the multilayer films prepared in Examples 1-2 and 1-3, surface morphological properties were confirmed using AFM. In general, as confirmed by the surface morphological properties, it is reported that the surfaces exhibiting the effect of preventing the non-selective adsorption of the cells exhibits a shape as if the fine particles are distributed on the measured surface as a whole.

Specifically, the microscope used in this experiment was using Digital Instrument's Dimension 3100 AFM, and the surface morphological properties were confirmed by taking a height image of AFM in a tapping mode using the AFM. It carried out by the method of checking the phase difference of a surface.

Example 2 -4: polymer electrolyte Multilayer film  Cell Adsorption Experiment of Film

Cell adsorption experiment of the polymer electrolyte multilayer membrane film was performed using the multilayer membrane.

In order to examine the interaction between the multilayered membrane and the cells, experiments were performed using several cells including kidney cells (HEK 293 kidney cells), and observed under a microscope.

Example  3: Multilayer  Check the effect

The film of PAH (8.5) / PMA (4.0) maintained about 40 °, but carboxylated beta After soaking in cyclodextrin once, it has a hydrophilic surface of about 20 ° .

On film Adsorbed carboxylated beta cyclodextrin silver Methylene blue  ( MeB ) With 0.001M complexation Can be formed.

The PAH / PMA film hardly reacted with MeB, but the film absorbed with Carboxylated beta cyclodextrin reacted with MeB to increase the UV absorption. It was found that the absorbance increased.

The effect of multi-layer thin films formed on various pH conditions on the host-guest interaction between carboxylated β-CyD and Methyleneblue (MeB) was investigated. Since PAH and PMA are weak polymer electrolytes, the charge density varies with pH, resulting in a change in the chain structure. Changes in the chain structure have various properties such as morphology and wettability of the film.

We formed various films under the conditions of PAH (4.0) / PMA (4.0), PAH (8.5) / PMA (4.0), PAH (8.5) / PMA (8.5). In addition, the experiment was performed to secure the space inside the film by treating with di - water of low pH (2.0) for 2 min after film formation .

In addition to MeB, Salicylic acid (SCA) was used as a guest molecule to test the interaction between Carboxylated β-CyD and Host-Guest.

Although the film became hydrophilic under the influence of Salicylic acid even without Carboxylated β-CyD, the contact angle decreased slightly due to the Host-Guest interaction of Carboxylated β-CyD with Salicylic acid.

Cell adsorption experiment

Renal cells (HEK 293 cells) are a species of cells that are useful in medicine and biomaterials because of their rapid proliferation and ease of culture. Substrate for cell adsorption experiment was coated with a multilayer thin film of two-thirds of 60mm diameter TCPSTissue Culture Grade Polystyrene Dish to compare the coated and unbarred areas (bare) It was. For the experiment cells give made of 37 ℃ put in a constant temperature bath to give a match to the culture media for the temperature suitable for the CO ₂ cells To divide the cells cultured in the incubator into the specimen, remove the media of TCPS growing, wash the cells of TCPS twice with DPBS (Dulbecco Phosphate Buffered Saline) and insert Tripsinf-EDTA 700Чl into TCPS. After wetting about 2 ~ 3 times horizontally, we tried to completely remove Trypsin-EDTA using Micro-Pipet. For stabilization of the cells, incubate for 5 minutes in the incubator and take out again to disperse 4ml of new media in the cells. Cells were dispersed with 1 ml of 4 ml of media and 4 ml of media was put on 70% ethanol sterilized sample. And again CO ₂ After incubation in the incubator and observed the adsorption process of the cells 1, 3, 5 days after the experiment. Cell experiments were performed depending on the presence or absence of carboxylated β-CyD.

2-2 Cationic β-CyD polymer synthesis

1) Experiment

1-1) Reagent

β-CyD, Epichlorohydrin (EP) and choline chloride (CC) were used by Aldrich. Dialysis was performed using Spectrum's molecular weight cut-off 1000 membrane. The entire process of dialysis and experiment was made with deionized water (> 18MΩcm Millipore Milli-Q).

1-2) Cationic β-CyD polymer [P (βCyD)] synthesis

Synthesis proceeded at a molar ratio of β-CyD / EP / CC = 1/15/1. Dissolve 1 g NaOH in 20 ml deionized water. 1.135g β-CyD is added to an aqueous NaOH solution. NaOH aqueous solution containing β-CyD is stirred at 25 ° C for 24 hours. Add 0.140 g of CC to the aqueous solution and add 1.388 g of EP at a rate of about 0.1 ml / min. After all the EP is added, heat it to 60 ℃. Maintain 60 ℃ and 600rpm during the polymerization. After two hours, the polymerization was terminated by neutral aqueous solution by adding 3N aqueous hydrochloric acid solution. The solution is dialisis for 24 hours using a molecular weight cut-off 1000 membrane. The solution obtained through dialysis can obtain pure powder through lyophilization.

2) results

2-1) GPC

GPC was used to measure the molecular weight of the synthesized Cationic β-CyD polymer. GPC smaple was prepared at the concentration of 3mg / 1ml and GPC standard was the result using PEG. Mn was 2211.5 g / mol and Mw was 2306.4 g / mol D: 1.0429.

2-2) NMR

NMR was measured to analyze the synthesized cationic β-CyD polymer structure. Implemented on the NMR of JNM-AL400, it has a 400MHz resonance frequency. NMR solvent was used Al Drich's D ₂ O.

2-3) Multilayer Thin Film Formation Using P (βCyD)

1. Experiment

1-1) Reagent

Poly (Acrylic acid) (Mw = 90.000) was used as a reagent of Polyscience. The polymer electrolyte was used without further purification. Dulbecco Phosphate Buffered Saline (DPBS), 0.25% Tripsinf-EDTA, and Dulbecco modified eagle medium (DMEM) were used for cell experiments. In addition, Gibco fetal bovine serum (FBS) and Antibioic-Antimycotic were used to make DMEM.

2. Results

After fixing the pH of P (βCyD) to 7.0, adjust the PAA pH to 3.0, 5.0, and 7.0, respectively.

After the film was formed and dyed with MeB, UV was measured.

Contact angle is the angle between the solid surface and the liquid surface when the liquid is thermodynamically balanced on the solid surface. At this time, since the angle varies depending on the state of the solid surface and the type of liquid, the contact angle shows the surface energy, the uniformity of the surface, and the degree of hydrophilicity. The contact angle was measured to evaluate the change in wettability as the film was laminated. In the wettability test, the contact angle of water was measured by using CA-A Contact angle analyzer of Face Company. Deionized water was used for the contact angle measurement, and a drop of 15 Чl was formed using a syringe, and then the measurement plate on which the film was lifted was slowly raised to contact the water drop, thereby preventing the influence of gravity on the contact angle measurement. First, the static contact angle was measured, and in that state, 15 ll was further added to measure the advancing contact angle. Finally, 15 Чl was sucked again from the droplet of 30 Чl to measure the Receding contact angle. The result value calculated | required the average value of the value measured 5 times by dropping a water droplet on the surface of a film.

In order to sufficiently charge density, the pH of P (βCyD) was adjusted to low pH. P (βCyD) adjusted to pH3.0 formed a film by PAA pH3.0, 5.0 7.0 and LBL method as in the previous experiment. This film is complexed with P (βCyD) in MeB. Observation was made using UV. Also compared with films of PAH / PAA, PAH / PAA / P (βCyD) / PAA.

If the guest molecules of the optically inactive form Cyclodextrin complexation with, is to have a characteristic which is active. Optical activity is induced by the interaction of cyclodextrin and guest molecules. Complexation can be confirmed by circular dichrorism spectra and structural information can be obtained. When complexation with MeB was found to have a CD value

1 is a schematic view showing a method of manufacturing a polymer electrolyte multilayer film including cyclodextrin in a polymer electrolyte multilayer film according to an embodiment of the present invention.

Figure 2 is a graph showing the results of the contact angle experiment showing the wettability of the polymer electrolyte multilayer according to an embodiment of the present invention.

3 to 18 are graphs or photographs showing contact angles, values measured through UV, and cell adhesion of the multilayer film according to an embodiment of the present invention.

Claims (9)

A polymer electrolyte multilayer membrane comprising cyclodextrin or a derivative thereof in an alternating stacked structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer. The method of claim 1, The anionic polymer electrolyte layer is one selected from the group consisting of polymethacrylic acid, polyacrylic acid and hyaluronic acid, and the cationic polymer electrolyte layer is any selected from the group consisting of polyallylamine hydrochloride and polyacrylamide. Multilayered film that consists of one. The method of claim 1, A multilayer membrane having at least two anionic polymer electrolyte layers and a cationic polymer electrolyte layer. The method of claim 3, The anionic polymer electrolyte layer is made of the same or different polymers, the cationic polymer electrolyte layer is made of the same or different polymers. The method of claim 1, Wherein said cyclodextrin is alpha-cyclodextrin or beta-cyclodextrin. An alternating stack structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer, One or both layers of the outermost layer of the alternating layered structure is composed of a cyclodextrin or a derivative thereof. The method of claim 6, The anionic polymer electrolyte layer is one selected from the group consisting of polymethacrylic acid, polyacrylic acid and hyaluronic acid, and the cationic polymer electrolyte layer is any selected from the group consisting of polyallylamine hydrochloride and polyacrylamide. Multilayered film that consists of one. The method of claim 6, Wherein said cyclodextrin is alpha-cyclodextrin or beta-cyclodextrin. A biosensing device comprising the multilayer film of claim 1.

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