Resin for preparing biochip, biochip and application

Abstract

The invention provides a resin for preparing a biochip, the biochip and application of the biochip. The resin provided by the invention comprises the following components: a first monomer, a second monomer, a third monomer, a first initiator and a second initiator, two or three different types of monomers are used at the same time, and a two-step polymerization method of photo-initiation and thermal-initiation polymerization is adopted, so that the obtained cured resin has good biocompatibility, hydrophilicity and anti-swelling performance, and is suitable for large-scale production. And due to the characteristic of long-term storage at room temperature, the defect that a gel micro-material is easy to volatilize and dry is overcome. The resin provided by the invention is used for manufacturing a biochip, the chip can show good low-adsorption characteristic without post-treatment, and the preparation method is simple, low in cost and suitable for mass production and application.

Description

Technical Field

[0001] The present invention relates to the technical field of biochips, and in particular to a resin for preparing biochips, the biochips and their applications. Background Art

[0002] The continuous progress of chip technology has provided a powerful tool for the miniaturization, high throughput and automation of biomedical analysis and detection. Currently, commonly used microfluidic polymer materials include polymethacrylate (PMMA), polydimethoxysilane (PDMS), hydrogel, etc. Biochips made of PMMA and PDMS usually need to be hydrophilic, which is complicated to operate and may have problems such as inconsistent hydrophilicity of various parts of the chip after hydrophilic treatment; chips made of hydrogel have good hydrophilicity, but poor mechanical properties. At room temperature, water is easily volatile, which affects the microstructure of the chip, so it is difficult to store for a long time at room temperature. Therefore, it is of great application value to develop a resin with good biocompatibility, hydrophilicity, low adsorption and other characteristics and use it to manufacture chips. Summary of the invention

[0003] In view of the shortcomings of the prior art mentioned above, the purpose of the present invention is to provide a resin for preparing a biochip, a biochip and its application, by initiating two-step polymerization of the resin by light and heat to obtain a resin with good biocompatibility, hydrophilicity and low adsorption properties. The prepared biochip can be stored at room temperature for a long time and will not absorb water and swell during use, thus overcoming the problem that hydrogel chips are easily volatile at room temperature and have harsh storage conditions.

[0004] To achieve the above-mentioned purpose and other related purposes, the present invention adopts the following technical solutions:

[0005] The first aspect of the present invention provides a resin for preparing a biochip, wherein the resin is prepared from the following components: a first monomer, a second monomer, a third monomer, a first initiator and a second initiator; the mass ratio of the first monomer, the second monomer and the third monomer is 1:0.2-5:0-0.05; the mass ratio of the first initiator, the second initiator and the total mass of the monomers is 0.001-0.1:0.001-0.01:1; the first monomer is selected from an acrylate derivative of polyethylene glycol; the second monomer is selected from acrylate compounds; the third monomer is selected from one or more of methoxypolyethylene glycol acrylate compounds, 2-methacryloyloxyethyl phosphorylcholine and methacrylic acid sulfobetaine.

[0006] The second aspect of the present invention is a method for preparing the above-mentioned resin for preparing biochips, comprising the following steps: uniformly mixing the first monomer, the second monomer, the first initiator or the first monomer, the second monomer, the third monomer, and the first initiator, adding an organic solution dissolved with the second initiator, and mixing thoroughly to obtain the resin for preparing biochips.

[0007] The third aspect of the present invention provides a biochip, which is prepared using the above-mentioned resin for preparing biochips as a substrate.

[0008] The fourth aspect of the present invention provides a method for preparing the above-mentioned biochip, comprising the following steps:

[0009] (1) forming the resin for preparing the biochip, and curing it with ultraviolet light to obtain a semi-finished biochip;

[0010] (2) The semi-finished biochip product is subjected to heat treatment and cooled to obtain a biochip.

[0011] A fifth aspect of the present invention provides the use of the above-mentioned biochip in biomedical analysis.

[0012] The resin for preparing a biochip and the biochip and their application provided by the present invention have the following beneficial effects:

[0013] 1. The resin provided by the present invention uses two or three different types of monomers at the same time and adopts a two-step polymerization method of light-initiated and heat-initiated polymerization. The obtained cured resin has good biocompatibility, hydrophilicity, anti-swelling properties, and can be stored for a long time at room temperature, overcoming the disadvantage of easy volatile drying of gel micromaterials.

[0014] 2. The present invention provides a resin for manufacturing chips. The obtained chip has good mechanical properties and can show good low adsorption characteristics without post-treatment. It is suitable for application in biomedical analysis, especially providing great convenience for constructing organoid models. The biochip preparation method provided by the present invention is simple, low-cost, and suitable for mass production and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Shown is a schematic diagram of the process of preparing a microarray biochip using the micro-nanoimprinting method of the present invention.

[0016] Figure 2 The morphology of the biochips prepared in Example 1, Comparative Example 1 and Comparative Example 2 of the present invention after being immersed in 37°C PBS (10mM, pH=7.4) for 48 hours is shown; a is the morphology of the biochip in Example 1 after immersion, b is the morphology of the biochip in Comparative Example 1 after immersion, and c is the morphology of the biochip in Comparative Example 2 after immersion. .

[0017] Figure 3 Shown are the morphology images of the biochips prepared in Example 1 of the present invention and Comparative Example 3 after being immersed in 37°C PBS (10mM, pH=7.4) for 48 hours; wherein a is the morphology image of the biochip in Example 1 after immersion, and b is the morphology image of the biochip in Comparative Example 3 after immersion.

[0018] Figure 4 Shown is a cell distribution diagram of an organoid model constructed using the biochip prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0019] The inventors of the present invention combined photo-induced polymerization with thermal-induced polymerization and selected the types of monomers to obtain a cured resin with good biocompatibility, hydrophilicity and anti-swelling properties through two-step polymerization. The cured resin was used to prepare chips, which showed good low adsorption properties. The present invention was completed on this basis.

[0020] The first aspect of the present invention provides a resin for preparing a biochip, wherein the resin is prepared from the following components: a first monomer, a second monomer, a third monomer, a first initiator and a second initiator; the mass ratio of the first monomer, the second monomer and the third monomer is 1:0.2-5:0-0.05; the mass ratio of the first initiator, the second initiator and the total mass of the monomers is 0.001-0.1:0.001-0.01:1; the first monomer is selected from an acrylate derivative of polyethylene glycol; the second monomer is selected from acrylate compounds; the third monomer is selected from one or more of methoxypolyethylene glycol acrylate compounds, 2-methacryloyloxyethyl phosphorylcholine and methacrylic acid sulfobetaine.

[0021] The material prepared entirely from the first monomer, the acrylate derivative of polyethylene glycol, has good biocompatibility and hydrophilicity, but due to the high hydrophilicity, the material easily absorbs water and swells in the water environment, resulting in damage to the material body. The material prepared entirely from the second monomer, the acrylate compound, exhibits certain hydrophobicity, and the surface of the chip composed of its polymer usually requires hydrophilic treatment. The present invention prepares a biochip with good biocompatibility, hydrophilicity and anti-swelling properties by adjusting the ratio of the first monomer to the second monomer, and the addition of the third monomer can further form a layer of hydrophilic polymer on the chip surface, which is beneficial to reduce the chip's adsorption performance to cells.

[0022] The present invention uses a two-step polymerization method of photo-initiated polymerization and thermal-initiated polymerization. The photoinitiator quickly initiates free radical polymerization of monomers under ultraviolet light irradiation to form a polymer cross-linked network to obtain a chip semi-finished product. Due to the short photopolymerization time and fast speed, the internal stress of the chip is relatively large, and there are problems such as monomer residue. By placing the chip semi-finished product in a thermal environment, the thermal initiator triggers the residual monomer or functional group to further react, which can improve the mechanical properties of the material and reduce the residual monomer; at the same time, the thermal environment releases the stress inside the material and improves the stability of the material.

[0023] In some embodiments of the present invention, the acrylate derivative of polyethylene glycol includes polyethylene glycol diacrylate and / or polyethylene glycol dimethacrylate.

[0024] In some embodiments of the present invention, the molecular weight of the acrylate derivative of polyethylene glycol is 400-10000Da, which can be 400-500Da, 500-1000Da, 1000-2000Da, 2000-4000Da, 4000-6000Da, 6000-8000Da, or 8000-10000Da.

[0025] In some embodiments of the present invention, the acrylic acid ester compound includes any one or more of propyl acrylate, butyl acrylate, pentyl acrylate, hexyl acrylate, propyl methacrylate, butyl methacrylate, pentyl methacrylate, and hexyl methacrylate.

[0026] In some embodiments of the present invention, the methoxy polyethylene glycol acrylate compound includes methoxy polyethylene glycol monomethacrylate and / or methoxy polyethylene glycol monoacrylate.

[0027] In some embodiments of the present invention, the first initiator is a photoinitiator selected from any one of 2-hydroxy-2-methylpropiophenone, 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone and phenyl (2,4,6-trimethylbenzoyl) lithium phosphate (LAP).

[0028] In some embodiments of the present invention, the second initiator is a thermal initiator selected from any one of azobisisobutyronitrile, azobisisoheptanenitrile and dibenzoyl peroxide.

[0029] The second aspect of the present invention provides a method for preparing a resin for preparing a biochip, comprising the following steps: uniformly mixing a first monomer, a second monomer, and a first initiator or a first monomer, a second monomer, a third monomer, and a first initiator, adding an organic solution in which the second initiator is dissolved, and mixing thoroughly to obtain a resin for preparing a biochip.

[0030] In some embodiments of the present invention, the organic solvent of the organic solution is selected from any one of dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), and dioxane (1,4-dioxane).

[0031] In some embodiments of the present invention, the mass ratio of the second initiator in the organic solution to the volume ratio of the organic solution is 40-60 mg / mL.

[0032] In some embodiments of the present invention, the mixing is performed at room temperature. In the present invention, the room temperature is room temperature, usually 20-25°C.

[0033] In some embodiments of the present invention, the resin used to prepare the biochip needs to be stored away from light to avoid degradation or cross-linking of the resin caused by light. Preferably, the storage temperature should be lower than 25°C to avoid deterioration caused by excessive temperature; the humidity should be lower than 70% to avoid degradation of the resin performance caused by excessive humidity.

[0034] The third aspect of the present invention provides a biochip, which is prepared by using the resin for preparing a biochip as a substrate. From the experimental results recorded in the examples, it can be seen that the biochip provided by the present invention has good biocompatibility, hydrophilicity and anti-swelling properties, and has low adsorption characteristics.

[0035] The fourth aspect of the present invention provides a method for preparing the above-mentioned biochip, comprising the following steps:

[0036] (1) forming the resin for preparing the biochip, and curing it with ultraviolet light to obtain a semi-finished biochip;

[0037] (2) The semi-finished biochip product is subjected to heat treatment and cooled to obtain a biochip.

[0038] In some embodiments of the present invention, the preparation and molding adopts 3D printing, micro-nano imprinting, soft lithography, casting or injection molding.

[0039] In some embodiments of the present invention, the light source of ultraviolet light is an ultraviolet lamp with a wavelength of 365nm.

[0040] In some embodiments of the present invention, the heat treatment is specifically to place the semi-finished biochip product in a constant temperature box for treatment.

[0041] In some embodiments of the present invention, the heat treatment temperature is 60-80°C, which can be 60-65°C, 65-70°C, 70-75°C, or 75-80°C; the heat treatment time is 3-6h, which can be 3-4h, 4-5h, or 5-6h.

[0042] In some embodiments of the present invention, the cooling process further includes washing and drying. Preferably, the washing medium is deionized water; the drying condition is drying in an oven at 45-55°C for 2-4 hours, more preferably drying in an oven at 50°C for 3 hours.

[0043] The fourth aspect of the present invention provides the application of the above-mentioned biochip in biomedical analysis. The biochip provided by the present invention has good mechanical properties and can show good low adsorption characteristics without post-treatment, and is suitable for application in biomedical analysis, especially providing great convenience for building organoid models.

[0044] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0045] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, as those skilled in the art grasp the prior art and record the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0046] Unless otherwise specified, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in the relevant technical fields. Unless otherwise specified, the materials and equipment used in the present invention are all commercially available.

[0047] Example 1

[0048] (1) Preparation of resin: Weigh 1 g of polyethylene glycol diacrylate (Mw=400), 1 g of butyl methacrylate, and 0.02 g of 2-hydroxy-2-methylpropiophenone, mix well, and then add 0.2 mL of azobisisobutyronitrile DMSO solution (50 mg / mL). After thorough mixing, obtain the resin and store it in the dark for later use.

[0049] (2) Preparation of biochip: The preparation process is as follows Figure 1As shown, a certain amount of resin is applied to the surface of the glass sheet, and the polydimethoxysilane mold with the microstructure array is carefully imprinted on the resin surface, and the vacuum treatment is performed to expel the residual air so that the resin can fully infiltrate the mold. A 365nm ultraviolet lamp is used to vertically irradiate the resin from the back of the glass sheet for 60s to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the semi-finished biochip has a microstructure array complementary to the mold. The semi-finished biochip is placed in a 70℃ constant temperature box, and the azobisisobutyronitrile decomposes to initiate secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. After 4 hours, it is cooled to room temperature, and the biochip is obtained after washing with deionized water and drying.

[0050] Example 2

[0051] (1) Preparation of resin: Weigh 1 g of polyethylene glycol diacrylate (Mw=400), 1 g of butyl methacrylate, 0.1 g of methacrylic acid sulfobetaine, and 0.02 g of 2-hydroxy-2-methylpropiophenone, mix well, and then add 0.2 mL of DMF solution of azobisisobutyronitrile (50 mg / mL). After thorough mixing, obtain a resin and store it in the dark for later use.

[0052] (2) Preparation of biochip: Absorb a certain amount of resin and coat it on the surface of the glass slide, carefully imprint the polydimethoxysilane mold with the microstructure array on the resin surface, and perform vacuum treatment to expel the residual air so that the resin can fully infiltrate the mold. Use a 365nm ultraviolet lamp to vertically irradiate the resin from the back of the glass slide for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the biochip has a microstructure array complementary to the mold. Place the semi-finished biochip in a 70°C constant temperature box, and the decomposition of azobisisobutyronitrile triggers the secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. Stop heating after 4 hours, cool to room temperature, wash with deionized water, and dry to obtain the chip.

[0053] Example 3

[0054] (1) Preparation of resin: Weigh 1 g of polyethylene glycol dimethacrylate (Mw=550), 1 g of butyl methacrylate, 0.1 g of methacrylic acid sulfobetaine, and 0.02 g of 2-hydroxy-2-methylpropiophenone, mix well, and then add 0.2 mL of azobisisobutyronitrile DMSO solution (50 mg / mL). After thorough mixing, obtain the resin and store it in the dark for later use.

[0055] (2) Preparation of biochip: Absorb a certain amount of resin and coat it on the surface of the glass slide, carefully imprint the polydimethoxysilane mold with the microstructure array on the resin surface, and perform vacuum treatment to expel the residual air so that the resin can fully infiltrate the mold. Use a 365nm ultraviolet lamp to vertically irradiate the resin from the back of the glass slide for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the biochip has a microstructure array complementary to the mold. Place the semi-finished biochip in a 70°C constant temperature box, and the decomposition of azobisisobutyronitrile triggers the secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. Stop heating after 4 hours, cool to room temperature, wash with deionized water and dry to obtain a biochip.

[0056] Example 4

[0057] (1) Preparation of resin: Weigh 1 g of polyethylene glycol diacrylate (Mw=1000), 1 g of butyl methacrylate, 0.1 g of methacrylic acid sulfobetaine, and 0.02 g of 2-hydroxy-2-methylpropiophenone, mix well, and then add 0.2 mL of NMP solution of azobisisobutyronitrile (50 mg / mL). After thorough mixing, obtain the resin and store it in the dark for later use.

[0058] (2) Preparation of biochip: Absorb a certain amount of resin and coat it on the surface of the glass slide, carefully imprint the polydimethoxysilane mold with the microstructure array on the resin surface, and perform vacuum treatment to expel the residual air so that the resin can fully infiltrate the mold. Use a 365nm ultraviolet lamp to vertically irradiate the resin from the back of the glass slide for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the biochip has a microstructure array complementary to the mold. Place the semi-finished biochip in a 70°C constant temperature box, and the decomposition of azobisisobutyronitrile triggers the secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. Stop heating after 4 hours, cool to room temperature, wash with deionized water and dry to obtain a biochip.

[0059] Example 5

[0060] (1) Preparation of resin: Weigh 1 g of polyethylene glycol diacrylate (Mw=400), 1 g of propyl methacrylate, 0.1 g of methacrylic acid sulfobetaine, and 0.02 g of 2-hydroxy-2-methylpropiophenone, mix well, and then add 0.2 mL of azobisisobutyronitrile DMSO solution (50 mg / mL). After thorough mixing, obtain the resin and store it in the dark for later use.

[0061] (2) Preparation of biochip: Absorb a certain amount of resin and coat it on the surface of the glass slide, carefully imprint the polydimethoxysilane mold with the microstructure array on the resin surface, and perform vacuum treatment to expel the residual air so that the resin can fully infiltrate the mold. Use a 365nm ultraviolet lamp to vertically irradiate the resin from the back of the glass slide for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the biochip has a microstructure array complementary to the mold. Place the semi-finished biochip in a 70°C constant temperature box, and the decomposition of azobisisobutyronitrile triggers the secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. Stop heating after 4 hours, cool to room temperature, wash with deionized water and dry to obtain a biochip.

[0062] Example 6

[0063] (1) Preparation of resin: Weigh 1 g of polyethylene glycol diacrylate (Mw=400), 0.5 g of propyl methacrylate, 0.1 g of methacrylic acid sulfobetaine, and 0.015 g of 2-hydroxy-2-methylpropiophenone, mix well, and then add 0.2 mL of azobisisobutyronitrile DMSO solution (50 mg / mL). After thorough mixing, obtain the resin and store it in the dark for later use.

[0064] (2) Preparation of biochip: Absorb a certain amount of resin and coat it on the surface of the glass slide, carefully imprint the polydimethoxysilane mold with the microstructure array on the resin surface, and perform vacuum treatment to expel the residual air so that the resin can fully infiltrate the mold. Use a 365nm ultraviolet lamp to vertically irradiate the resin from the back of the glass slide for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the biochip has a microstructure array complementary to the mold. Place the semi-finished biochip in a 70°C constant temperature box, and the decomposition of azobisisobutyronitrile triggers the secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. Stop heating after 4 hours, cool to room temperature, wash with deionized water and dry to obtain a biochip.

[0065] Comparative Example 1

[0066] (1) Preparation of resin: 1 g of polyethylene glycol diacrylate (Mw=400), 1 g of butyl methacrylate, and 0.02 g of 2-hydroxy-2-methylpropiophenone were weighed and mixed thoroughly to obtain a resin, which was then stored in the dark for later use.

[0067] (2) Preparation of biochip: A certain amount of resin is applied to the surface of a glass sheet, and the polydimethoxysilane mold with a microstructure array is carefully pressed onto the resin surface, and then vacuumed to remove the residual air so that the resin can fully infiltrate the mold. A 365nm ultraviolet lamp is used to irradiate the resin vertically from the back of the glass sheet for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a microstructure array complementary to the mold is obtained, which is then washed with deionized water and dried to obtain a biochip, which is then stored for later use.

[0068] Comparative Example 2

[0069] (1) Preparation of resin: Weigh 1 g of polyethylene glycol diacrylate (Mw=400), 0.01 g of 2-hydroxy-2-methylpropiophenone and 0.1 mL of azobisisobutyronitrile DMSO solution (50 mg / mL), mix thoroughly to obtain a resin, and store in the dark for later use.

[0070] (2) Preparation of biochip: Absorb a certain amount of resin and coat it on the surface of the glass slide, carefully imprint the polydimethoxysilane mold with the microstructure array on the resin surface, and perform vacuum treatment to expel the residual air so that the resin can fully infiltrate the mold. Use a 365nm ultraviolet lamp to vertically irradiate the resin from the back of the glass slide for 60 seconds to quickly solidify the resin. After carefully peeling off the mold, a semi-finished biochip is obtained, and the surface of the biochip has a microstructure array complementary to the mold. Place the semi-finished biochip in a 70°C constant temperature box, and the azobisisobutyronitrile decomposes to initiate secondary polymerization of the resin, further improving the crosslinking degree and mechanical properties of the resin. Stop heating after 4 hours, cool to room temperature, wash with deionized water, and dry to obtain a biochip, which is stored for later use.

[0071] Comparative Example 3

[0072] (1) Preparation of resin: 1 g of methacryloyl-gelatin (GelMA) and 5 mg of ammonium persulfate (APS) were weighed and dissolved in 1 mL of an aqueous solution of 1% by mass of 2-hydroxy-2-methylpropiophenone to obtain a GelMA resin solution.

[0073] (2) Preparation of biochips: After a certain amount of resin solution is applied to the surface of a glass slide, a polydimethoxysilane mold with a microstructure array is imprinted on the resin surface and vacuumed to expel residual air so that the solution can fully infiltrate the mold. A 365nm UV lamp is used to irradiate vertically from the back of the glass slide for 60 seconds, and the solution quickly solidifies to form a hydrogel. After carefully peeling off the mold, place it in a 70°C constant temperature box, and APS initiates the secondary polymerization of the resin to further improve the crosslinking degree and mechanical properties of the hydrogel. After washing with deionized water and drying, the hydrogel microarray biochip is obtained and stored for later use.

[0074] Figure 1The main steps of preparing microarray chips by the micro-nano imprinting method of the present invention are given. Figure 1 It can be seen that the preparation method of the biochip provided by the present invention has few steps, is simple to operate, and can be completed using existing equipment.

[0075] Anti-swelling test: The biochips prepared in Example 1 and Comparative Examples 1 to 3 of the present invention were immersed in 37° C. PBS (10 mM, pH=7.4), and photographed after 48 hours to observe the morphological changes of the microarray region of the biochip before and after.

[0076] Figure 2 The morphology of the biochips prepared in Example 1, Comparative Example 1 and Comparative Example 2 of the present invention after being immersed in 37°C PBS (10mM, pH=7.4) for 48 hours. Figure 2 It can be seen that the biochip prepared in Example 1 ( Figure 2 a) The structure remains intact, while the comparative example 1 ( Figure 2 b) and Comparative Example 2 ( Figure 2 The biochips prepared in (c) all showed obvious swelling and damage.

[0077] Figure 3 The morphology of the biochips prepared in Example 1 and Comparative Example 3 of the present invention after being immersed in 37°C PBS (10mM, pH=7.4) for 48 hours. Figure 3 It can be seen that the biochip prepared in Comparative Example 3 ( Figure 3 The macroscopic morphology of the biochip (b) in Example 1 has no obvious change, but the microstructure can be observed to have swollen and deformed under a microscope. Figure 3 There is no obvious change in a), showing an inverted pyramid structure consistent with the mold. The above experimental results show that the resin provided by the present invention has good anti-swelling performance.

[0078] Cell experiment: The biochip was sterilized using ethylene oxide sterilization and then placed in a matching culture dish container. The microarray area of ​​the biochip was rinsed once with sterile PBS, and then 1 mL of lung tumor cell suspension (5×10 6 The cells were added to the microarray area of ​​the chip, and the chip was gently shaken and left to stand for 15 minutes to allow the cells to be evenly dispersed in the biochip. After the cells were cultured at 37°C and 5% CO2 for 48 hours, they were observed and photographed using a microscope.

[0079] Figure 4 This is a cell distribution diagram of an organoid model constructed using the biochip prepared in Example 1 of the present invention. Figure 4It can be seen that the cells are uniformly captured by the microarray on the surface of the biochip and grow well, indicating that the biochip has good cell compatibility. Due to the spatial limitations of the microarray, the cells aggregate into balls and gradually form organoids. In addition, the microarray exhibits obvious low adsorption characteristics, and the cells do not have obvious adhesion phenomenon.

[0080] In summary, the present invention successfully prepares a resin material and uses it to prepare a biochip, showing good biocompatibility, hydrophilicity and anti-swelling properties. The present invention effectively overcomes various shortcomings of the prior art and has high industrial utilization value.

[0081] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A resin for preparing a biochip, characterized in that: The resin comprises the following components: a first monomer, a second monomer, a third monomer, a first initiator and a second initiator; the mass ratio of the first monomer, the second monomer and the third monomer is 1:0.2-5:0-0.05; the mass ratio of the first initiator, the second initiator and the total mass of the monomers is 0.001-0.1:0.001-0.01:1; the first monomer is selected from acrylate derivatives of polyethylene glycol; the second monomer is selected from acrylate compounds; the third monomer is selected from one or more of methoxy polyethylene glycol acrylate compounds, 2-methacryloyloxyethyl phosphorylcholine and methacrylic acid sulfobetaine.

2. The resin for preparing a biochip according to claim 1, characterized in that: Includes one or more of the following characteristics: 1) The acrylate derivative of polyethylene glycol includes polyethylene glycol diacrylate and / or polyethylene glycol dimethacrylate; 2) The acrylic acid ester compound includes any one or more of propyl acrylate, butyl acrylate, pentyl acrylate, hexyl acrylate, propyl methacrylate, butyl methacrylate, pentyl methacrylate, and hexyl methacrylate; 3) The methoxy polyethylene glycol acrylate compound includes methoxy polyethylene glycol monomethacrylate and / or methoxy polyethylene glycol monoacrylate; 4) the first initiator is a photoinitiator, selected from any one of 2-hydroxy-2-methylpropiophenone, 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone and phenyl (2,4,6-trimethylbenzoyl) phosphate lithium salt; 5) The second initiator is a thermal initiator selected from any one of azobisisobutyronitrile, azobisisoheptanenitrile and dibenzoyl peroxide.

3. The resin for preparing a biochip according to claim 2, characterized in that: The molecular weight of the acrylate derivative of polyethylene glycol is 400-10000Da; and / or the molecular weight of methoxy polyethylene glycol monomethacrylate and methoxy polyethylene glycol monoacrylate in the third monomer is 1000-10000Da.

4. A method for preparing a resin for preparing a biochip according to claim 1, characterized in that: The method comprises the following steps: uniformly mixing the first monomer, the second monomer, the first initiator or the first monomer, the second monomer, the third monomer, the first initiator, adding an organic solution containing the second initiator, and fully mixing to obtain a resin for preparing a biochip.

5. The resin for preparing a biochip according to claim 4, characterized in that: The organic solvent of the organic solution is selected from any one of dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, and dioxane; and / or the mass ratio of the second initiator in the organic solution to the volume of the organic solution is 40-60 mg / mL.

6. The resin for preparing a biochip according to claim 4, characterized in that: The mixing is carried out at room temperature; And / or, the obtained resin for preparing the biochip needs to be stored away from light.

7. A biochip, characterized in that: The biochip is prepared by using the resin for preparing a biochip as described in any one of claims 1 to 6 as a substrate.

8. A method for preparing a biochip according to claim 7, characterized in that: The steps include: (1) forming the resin for preparing the biochip, and curing it with ultraviolet light to obtain a semi-finished biochip; (2) The semi-finished biochip product is subjected to heat treatment and cooled to obtain a biochip.

9. The method for preparing a biochip according to claim 8, characterized in that: Includes one or more of the following features: (a) The preparation and molding is carried out by any one or more of 3D printing, micro-nano imprinting, soft lithography, casting or injection molding methods; (b) the light source of the ultraviolet light is an ultraviolet lamp with a wavelength of 365 nm; (c) The heat treatment specifically comprises placing the semi-finished biochip product in a constant temperature box for treatment; (d) the heat treatment temperature is 60 to 80° C. and the heat treatment time is 3 to 6 hours; (e) After cooling, it also includes cleaning and drying.

10. Use of the biochip according to claim 7 or the biochip prepared according to any one of claims 8 to 9 in biomedical analysis.

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