KR100729953B1 - Method for manufacturing plastics substrate by plasma process and plastics substrate manufactured using the same - Google Patents

Abstract

The present invention provides a method of manufacturing a plastic substrate having low self-luminescence and improved specificity, a) R _a <3 nm or R _q under a condition of 50 μm × 50 μm or less. Molding a plastic substrate having an Atomic Force Microscope (AFM) surface roughness of <4 nm; b) treating the plastic substrate with a plasma; And c) treating the substrate for surface modification monomers. According to the method of the present invention, there is provided a plastic substrate which can significantly reduce the self-luminescence and thereby significantly improve the detection specificity of the target biomolecule. Therefore, in spite of the advantages of various designs including microstructures compared to glass substrates, plastic substrates, which have been limited in use due to the disadvantage of high self-luminescence, can be widely used for microarrays, biochips or well plates. In addition, according to the present invention it is possible to improve the specificity of the substrate surface and to easily adjust the characteristics.

Plastic, Substrate, Plasma, Surface Modification, Self-luminous, Roughness, Microarray, Biochip, Well Plate

Description

Method for manufacturing plastics substrate by plasma process and plastics substrate manufactured using the same

1 schematically illustrates a plasma generating apparatus used for plasma processing in an embodiment of the present invention.

Figure 2 schematically shows the reaction taking place in the plasma treatment step and the allyl glycidyl ether treatment step as monomers for surface modification of the process according to the invention.

Figure 3a is a photograph of a plastic substrate according to the present invention, Figure 3b schematically shows a plastic substrate according to the present invention including a microfluidic structure.

4 illustrates the results of immobilizing the probe biomolecules using the plastic substrate according to the present invention, hybridizing the target biomolecules, and detecting them by fluorescence.

The present invention relates to a method for producing a plastic substrate having excellent detection specificity of a target biomolecule and a plastic substrate produced by the method.

Recently, with the development of biotechnology, nucleotide sequences and protein sequences constituting genetic information of individuals have been revealed. Accordingly, there is an active movement to develop biochips for the purpose of sequence analysis and disease diagnosis. The biochip is a high density integrated micromolecule such as DNA or protein to be analyzed on a substrate. Biochips are classified into photolithography chips, pin type spotting chips, or inkjet type spotting chips according to manufacturing methods.

The target biomolecule to be analyzed is coupled to the biochip in which the probe biomolecules are immobilized in a microarray form, and different binding degrees are detected according to the complementary degree of the probe biomolecule and the target biomolecule. The detection of the binding degree is generally performed by an optical method of hybridizing a target biomolecule labeled with a fluorescent material with a probe biomolecule, and then detecting a signal emitted from the fluorescent material.

The microarray technique is advantageous in that a large amount of biological information can be obtained by using very small amounts of biomolecules such as DNA or protein. Biochip is a lab-on-a-chip (LOC) concept that implements all pre-processing and analysis such as separation, purification and amplification of target biomolecules, including the fixation of probe biomolecules in microarray form. With the concept of point of care (POC), which enables immediate diagnosis at the emergency site, it is gradually becoming an e-Healthcare concept that can check and predict personal health conditions regardless of time and place through convergence with the information and communication field. It is developing.

In the power generation process, surface modification for immobilizing biomolecules on a specific substrate is the most basic technology, and a wet process is generally used for this purpose. In the conventional wet process, a silane compound having a functional group such as epoxide, amine or aldehyde is diluted with a solvent such as ethanol, coated on a glass substrate in the same manner as a dip coating, and then heat treated at a temperature of 100 ° C. or higher for about 1 hour. It is a method of manufacturing by. In this case, the material that can be used as the solvent may be selected from the group of alcohol solvents such as methanol, propanol and butanol, or methyl ethyl ketone or dimethylformamide in addition to ethanol. In addition, the coating method may be applied in various ways such as spray coating, spin coating in addition to the tip coating method.

However, the biggest technical limitation of the conventional wet process is that it is difficult to partially modify the surface of the micro size. Therefore, a new process technology that can overcome the technical limitations of the conventional wet process is increasingly required.

Meanwhile, various materials such as glass, silicon, or plastic are used as the substrate for the microarray. Among these materials, plastics have advantages in that they are easier to design various materials including microstructures, easier to mass-produce, and more economical than other materials, but have high self-luminescence and low detection specificity of target biomolecules. Because of this situation is not widely used.

The inventors of the present invention have repeatedly studied to solve the problems of the prior art, as a result of controlling the roughness of the plastic substrate during the manufacture of the plastic substrate and processing the plasma during surface modification, the self-luminescence is significantly reduced to detect the biomolecules. It was confirmed that the plastic substrate with excellent degree can be produced.

Accordingly, it is an object of the present invention to provide a method for producing a plastic substrate which is capable of reducing self-luminescence and improving detection specificity of biomolecules and easily controlling properties.

In addition, another object of the present invention is to provide a plastic substrate produced by the method, which reduces the self-luminescence and improves the detection specificity of biomolecules.

In order to achieve the object of the present invention, the present invention provides a method for producing a plastic substrate, the present invention is a) R _a <3 nm or R _q under the condition of 50 ㎛ × 50 ㎛ Molding a plastic substrate having an Atomic Force Microscope (AFM) surface roughness of <4 nm; b) treating the plastic substrate with a plasma; And c) treating the substrate with a surface modification monomer.

In order to achieve the another object of the present invention, the present invention has an AFM (Atomic Force Microscope) surface roughness of R _a <3 nm or R _q <4 nm under conditions of 50 ㎛ × 50 ㎛, plasma and surface Provided is a plastic substrate characterized in that the monomer for modification has been treated.

Hereinafter, a method of manufacturing a plastic substrate according to the present invention will be described step by step.

Method of manufacturing a plastic substrate according to the present invention includes the step of molding a plastic substrate having a R _a <3 nm or surface roughness R _q <4 nm in (Atomic Force Microscope) AFM under the following conditions 50 ㎛ × 50 ㎛ .

The plastic substrate according to the present invention is selected from the group consisting of polystyrene (PS), cycloolefin copolymer (COC) and polycarbonate (PC), or a transparent resin selected from the group consisting of polypropylene (PP) and polyethylene (PE). Translucent resin. Preferably, the plastic substrate may be the transparent resin. The plastic substrate should have chemical resistance and heat resistance.

In the embodiment of the present invention, a PS resin that satisfies high economic efficiency, low shrinkage ratio, and adequate heat resistance was used. The factors affecting the self-luminescence of the plastic substrate are not only due to the resin itself and the additives, but also sensitive to local thermal decomposition, residual stress, and mold roughness in the processing step.

Plastic substrates with low surface roughness were found to significantly reduce self luminescence (see Example 1). Specifically, the plastic substrate according to the present invention preferably has an atomic force microscope (AFM) surface roughness of R _a <3 nm or R _q <4 nm under a condition of 50 μm × 50 μm or less.

In the above, R _a represents average roughness and R _q represents root-mean-square roughness.

The plastic substrate having a low surface roughness according to the present invention can be produced, for example, by an injection molding method. That is, a plastic substrate having a low surface roughness may be manufactured by injection molding the plastic substrate using a mold mirrored to an optical lens level.

Next, the method of manufacturing a plastic substrate according to the present invention includes the step of treating a plasma on the plastic substrate.

In the present invention, the plasma is preferably an argon plasma. Nitrogen or oxygen plasmas are somewhat inappropriate because they form not only radicals but also various nitrogen or oxygen compounds on the substrate surface.

The plasma energy density, which defines the nature of the plasma, is expressed as the ratio of W / FM, where W is the unit of J / sec by the power applied from the plasma generator, F is the unit of mole / sec by the flow rate, and M Represents the unit of g / mole in molecular weight. That is, the denominator has a mass flow rate unit in g / sec, and the whole has a unit in J / g.

In the present invention, the plasma is preferably a continuous plasma or pulsed plasma having an energy density of 10 ⁸ J / kg or less. Pulsed plasma has the advantage of minimizing functional damage compared to continuous plasma.

In the present invention, the plasma is preferably treated for 10 minutes or less.

In the present invention, the frequency of the plasma is preferably microwave (2.45 GHz) or RF (13.56 MHz).

The plasma treatment according to the present invention can be performed by conventional methods or apparatus. 1 schematically illustrates a plasma generating apparatus used for plasma processing in an embodiment of the present invention.

The plasma process used in the present invention belongs to the low temperature / vacuum plasma group, and since the damage probability such as deformation due to temperature is very small in the process, a material such as plastic is also possible. Therefore, it is advantageous to use plastics instead of inorganic substrates such as glass and silicon. 3b is because plastic is easy to mass-produce and economical with products having microstructures compared to glass or silicon. In addition, since the plasma process is a dry process under vacuum conditions, the surface may be treated very uniformly, and a different surface modification may be performed by using a mask having a micro pattern.

Next, the method of manufacturing a plastic substrate according to the present invention includes the step of treating the surface-modifying monomer to the substrate.

In the present invention, the monomer to be treated is selected from the group of chemicals containing an epoxide functional group, preferably allyl glycidyl ether of Formula 1 or glycid of Formula 2 having a carbon double bond Glycidyl methacrylate, whereby the plastic surface may be modified with epoxide groups. Allyl glycidyl ethers are preferred over glycidyl methacrylate in terms of reducing the extent of self-luminescence increase.

<Formula 1>

<Formula 2>

In addition, the monomer to be treated has a high breaking point such as hexanal, octanal, nonanal, decanal, dodecyl aldehyde, trans-2-pentenal, trans-2-hexen, etc. when the substrate is modified with an aldehyde group. Compound is preferred.

In addition, the monomer to be treated is (3-aminopropyl) trimethoxysilane, 3- (diethylamino) propylamine, propylamine, butylamine, 1-amino-2-propanol, when the substrate is modified with an amine group. 3-methoxy, which is an allyl amine or ether compound having a carbon double bond, can be selected from the group of compounds containing primary amine functional groups such as tris (2-aminoethyl) amine, isobutylamine, isopentylamine, etc. Propyl amine is more preferred.

In the present invention, the plasma treatment step and the surface treatment monomer treatment step may be performed separately, but may be performed simultaneously.

Figure 2 schematically shows the reaction taking place in the plasma treatment step and the allyl glycidyl ether treatment step as monomers for surface modification of the process according to the invention. This process introduces an epoxide functional group to the substrate surface.

Referring to Figure 2 (a), when the substrate is treated with an argon plasma, the plasma does not chemically bond to the substrate, but only forms radicals. Referring to Figure 2 (b), the carbon double bond of the radical and the monomer formed above is reacted, and the radical is transferred to form compounds of various structures. The most important feature of the present invention is that, through the plasma process step and condition control, the compound structure can be derived in a direction that can contribute to specificity improvement. Referring to Figure 2 (c), hydrogen is introduced to terminate the radicals.

In the method of manufacturing a plastic substrate according to the present invention, the method may further include forming a microfluidic structure on the plastic substrate. The step may, for example, be carried out simultaneously with the molding step of the plastic.

In another aspect of the invention, the invention provides a plastic substrate made by the method according to the invention.

The plastic substrate according to the present invention has an AFM (Atomic Force Microscope) surface roughness of R _a <3 nm or R _q <4 nm under a condition of 50 μm × 50 μm or less, and the surface is treated with a monomer for plasma and surface modification. It is characterized by.

The plasma may be an argon plasma. In addition, the plasma may be a continuous plasma or a pulsed plasma having an energy density of 10 ⁸ J / kg or less. In addition, the plasma may be treated for 10 minutes or less. In addition, the frequency of the plasma may be microwave (2.45 GHz) or RF (13.56 MHz).

Preferably, the plastic substrate is selected from the group consisting of polystyrene (PS), cycloolefin copolymer (COC) and polycarbonate (PC), or a transparent resin selected from the group consisting of polypropylene (PP) and polyethylene (PE). The translucent resin selected may be.

The monomer to be treated is an allyl glycidyl ether or glycidyl methacrylate containing an epoxide functional group and having a carbon double bond, whereby the plastic substrate surface is an epoxide group. Can be modified.

Alternatively, the monomer to be treated is hexanal, octanal or nonanal with a boiling point of 80 ° C. or higher, whereby the plastic substrate surface can be modified with an aldehyde group. .

Alternatively, the monomer to be treated is allylamine or an ether compound, 3-methoxypropylamine, which contains a primary amine functional group and has a carbon double bond, whereby the surface of the plastic substrate is The amine group can be modified.

Figure 3a is a photograph of a plastic substrate according to the present invention, Figure 3b schematically shows a plastic substrate according to the present invention including a microfluidic structure.

4 illustrates the results of immobilizing the probe biomolecules using the plastic substrate according to the present invention, hybridizing the target biomolecules, and detecting them by fluorescence.

In the embodiment of the present invention, the probe biomolecules were immobilized on the conventional glass substrate and the plastic substrate prepared by the method according to the present invention, hybridized with the target biomolecules, and compared with the results detected by fluorescence. As a result, the specificity of the plastic substrate according to the present invention was remarkably superior to the conventional substrate.

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

<Example 1>

Self-luminescence measurement according to surface roughness

In order to check the effect of surface roughness of the plastic substrate on the self-luminescence, the plastic was molded from the PS 15NFI resin produced by LG Chem using a mold mirrored to a normal level and a mold mirrored to an optical lens level.

The roughness of the plastic substrates manufactured in each mold was measured by using an atomic force microscope (AFM), and R _a was 3 nm or R _q was 4 nm under the condition of 50 μm × 50 μm, depending on the mirror processing level. It was.

The self-luminescence measurement was performed using an Axon GenePix 4000B scanner, and the self-luminescence measurement conditions were 100% laser power and PMT tube level 600.

The results are shown in Table 1. As shown in Table 1, the surface roughness improvement was able to lower the self-luminescence to about 26% for 532 nm laser and about 44% for 635 nm laser.

From the above results, it was confirmed that the self-luminescence of the plastic substrate can be reduced by lowering the roughness of the plastic substrate.

TABLE 1

Mold mirroring level Intensity @ 532 nm Intensity @ 635 nm General level 2992 442 Optical lens level 782 194

<Example 2>

Self-luminescence measurement according to PS resin

In order to confirm self-luminescence according to the PS resin, three types of PS resins in mass production, namely, LG Chem PS 15NFI, LG Chem PS 25SPI and BASF PS 147F, were compared. In this case, the mold used was a mold mirror-treated at the optical lens level, and the self-luminescence measurement conditions were the same as in Example 1.

The results are shown in Table 2. As shown in Table 2, the three types of PS resin showed a significant difference in self-luminescence, and LG Chem PS 15NFI showed the best results. This may be due to the combined contribution of the additives contained in the PS resin and the local pyrolysis and residual stress in the processing step. In particular, when comparing LG Chem PS 15NFI and 25SPI with similar additive content and composition, 15NFI is low in molecular weight and excellent in fluidity, so there is little possibility of local pyrolysis and residual stress.

TABLE 2

PS resin Intensity @ 532 nm Intensity @ 635 nm LG Chemical PS 15NFI 782 194 LG Chemical PS 25SPI 1558 308 BASF PS 147F 1800 747

<Example 3>

Measurement of self-luminescence increase with plasma treatment time

As in the above embodiment, the level of self-luminous emission with plasma treatment time was confirmed for the plastic substrate manufactured by using a mold processed by mirroring LG Chem PS 15NFI resin to the optical lens level.

The plasma generating apparatus used for the plasma treatment is schematically shown in FIG. The argon flow rate was fixed at 30 cc / min, the pressure at 20 Pa, and the plasma power at 100 W, and were treated for up to 30 minutes at 5 minute intervals, respectively. The plasma energy density was 1.12 × 10 ⁸ J / kg under the above conditions. The self-luminescence measurement conditions were the same as in Example 1.

The results are shown in Table 3. As shown in Table 3, the linear increase was about 110 levels per minute depending on the treatment time.

TABLE 3

Processing time 5 minutes 10 minutes 15 minutes 20 minutes 25 minutes 30 minutes Intensity @ 532 nm 1615 2253 2627 3171 3551 4641

<Example 4>

Measurement of self-luminescence increase according to plasma power

Using a plastic substrate manufactured in the same manner as in Example 3, the level of self-luminescence increase according to plasma power was determined. The argon flow rate was fixed at 30 cc / min, the pressure was 20 Pa, and the treatment time was 10 minutes, and each treated up to 300 W at 50 W intervals. The plasma energy density was 5.61 × 10 ⁷ J / kg to 3.36 × 10 ⁸ J / kg under the above conditions. The self-luminescence measurement conditions were the same as in Example 1.

The results are shown in Table 4. As shown in Table 4, the linear increase was about 110 levels per 10 W depending on the plasma power.

The result of Example 3 and Example 4 was summed up, and the same self-luminescence was shown because the amount of energy (6 kJ) was the same when treated at 100 W for 1 minute and at 10 W for 10 minutes.

TABLE 4

Plasma power 50 W 100 W 150 W 200 W 250 W 300 W Intensity @ 532 nm 1666 2253 2901 3092 3600 4239

Example 5

Measurement of increase in self-luminescence with pressure

Using a plastic substrate manufactured in the same manner as in Example 3, the level of self-luminescence increase according to pressure was determined. The argon flow rate was fixed at 30 cc / min, plasma power 100 W, and the treatment time was 10 minutes, and each treated up to 100 Pa at 20 Pa intervals. Under the above conditions, the plasma energy density was 1.12 × 10 ⁸ J / kg, and the self-luminescence measurement conditions were the same as in Example 1.

The results are shown in Table 5. As shown in Table 5, the increase in self-luminescence with pressure was very small. Low temperature / vacuum plasma, such as the method used in the present invention, is characterized in that no plasma is generated when the pressure is increased.

TABLE 5

pressure 20 Pa 40 Pa 60 Pa 80 Pa 100 Pa Intensity @ 532 nm 2253 2364 2669 2615 2620

<Example 6>

Improved specific sequence biomolecule specificity

A plasma process was applied to the plastic substrate manufactured in the same manner as in Example 3, and the surface was modified to a surface having an epoxide functional group.

Plasma treatment conditions are divided into two stages, and in the first stage, radicals are introduced into the plastic substrate using an argon plasma. An argon flow rate of 20 cc / min, a pressure of 20 Pa, a plasma power of 100 W, and a treatment time of 1 minute were applied. At this time, the plasma energy density was 1.68 × 10 ⁸ J / kg.

In the second step, the allyl glycidyl ether monomer and argon were treated with a plasma. The argon flow rate and pressure were the same as the first step, and the treatment time was 5 minutes. In this case, in order to minimize functional group damage and increase in self-luminescence, plasma power 100 W was treated in a pulsed manner. The pulse periods were set to 1 second, 2 seconds, 5 seconds, and infinity, respectively, and turned on only for 100 msec. When the pulse period is 1 second, it is turned off for 900 msec and turned on for 100 msec. Finally, by ending the radical with hydrogen, an epoxide plastic substrate was produced.

Two kinds of probe biomolecules (SEQ ID NOs: 1 and 2) were immobilized on the prepared epoxide plastic substrate using Perkin-Elmer's piezoelectric type microarrayer. The buffer used was 3 × SSC, 0.01% SDS, and reacted for 16 hours at 60 ℃ temperature and 70% relative humidity conditions. Then, the mixture was reacted for 30 minutes in a 50 mM Tris-HCl (pH 9.0) solution at a temperature of 50 ° C, washed twice with 0.2% SDS for 2 minutes, washed twice with distilled water for 2 minutes, and centrifuged at 1,000 rpm for 5 minutes. Dried.

On the other hand, the target biomolecule is a PCR product to which the fluorescent material is bound, mixed with a hybridization solution (3 × SSC, 0.001% SSC) in a ratio of 1: 20 and reacted for 30 to 60 minutes at a temperature of 55 ℃. Then, the mixture was washed for 5 minutes at room temperature with 1 x SSC, for 2 minutes at 0.1 x SSC, and dried at room temperature. Fluorescence measurement conditions were the same as the self-luminescence measurement conditions of Example 1.

Table 6 shows the results of comparing the specificity of two biomolecules having similar nucleotide sequences that are difficult to distinguish from each other. The specificity is given by

Specificity = (fluorescence amount of target A1 reacted with probe A1) / (fluorescence amount of target A1 reacted with probe A2)

As a control, a comparison was made using a commercially available epoxide glass substrate made by a conventional wet process.

As shown in Table 6, by increasing the pulse period of the plasma in the method according to the present invention, it was possible to increase the specificity for similar biomolecules that were difficult to distinguish.

TABLE 6

division Control Cycle: 1 second Cycle: 2 seconds Cycle: 5 seconds Cycle: Infinity Specificity 1.3 1.8 5.3 6.3 7.2

<Example 7>

Specificity comparison

Specificity was compared for the plastic substrates and controls according to the present invention with respect to biomolecules B, C and D with good specificity using the method described in Example 6 (SEQ ID NOs: 3, 4 and 5, respectively). The plastic substrate used in this example was manufactured in the same manner as in Example 6 except that the pulse period of the plasma was set to 3 seconds. A conventional epoxide glass substrate was used as the control.

Specificity results confirmed above are shown in Table 7. As shown in Table 7, the specificity of the plastic substrate according to the present invention was maintained at about two times higher than that of the conventional glass substrate, and in particular, showed excellent performance with respect to low concentration of target biomolecules.

TABLE 7

division Target B Target C Target D Density: 100 copy Density: 100,000 copy Plastic substrate of the present invention 84 16 33 67 Control 20 9 6 24 Improvement rate 4.2 times 1.8 times 5.5 times 2.8 times

<Example 8>

Specificity comparison

In a similar manner to Example 6, the plastic substrate was modified to a surface having an aldehyde functional group. The monomer used was octanal and the pulse period was 0.5 seconds.

 As the control group, an aldehyde plastic substrate of Greiner Bio-One using a conventional aldehyde glass substrate and a wet process was used.

Probe biomolecules E, F, G, and F1 (SEQ ID NOs: 6, 7, 8, and 9, respectively) were fixed on the produced aldehyde plastic substrate and two control substrates in the same manner as in Example 6. However, due to the nature of the aldehyde substrate, a reduction reaction is required, which was reacted for 10 minutes in a solution of 0.625 g NaBH ₄ / (120 mL 100% EtOH + 375 mL 1X PBS).

As shown in Table 8, the specificity of the plastic substrate according to the present invention maintained a value equivalent to that of the conventional glass substrate, and showed superior performance with respect to the plastic substrate manufactured by the conventional wet process.

Specificity for target F is the amount of fluorescence of target F reacted with probe biomolecule of SEQ ID NO: 7 divided by the amount of fluorescence of target F reacted with probe biomolecule of SEQ ID NO: 9, which is why the value is generally low This is because the base sequences are similar to each other, so it is difficult to distinguish them.

TABLE 8

division Target E Target F Target G Plastic substrate of the present invention 28 3 29 Control (Glass) 31 2 30 Control (Plastic) 10 1.5 9

Example 9

Specificity comparison

In a similar manner to Example 6, the plastic substrate was modified to a surface having an amine function. The monomer used was 3-methoxypropyl amine and the pulse period was 5 seconds. An amine plastic substrate of Greiner Bio-One, which applied a wet process, was used as the control.

Probe biomolecules H, I, and J (SEQ ID NOs: 10, 11, and 12, respectively) were fixed in a similar manner as in Example 6. The probe biomolecule was a PCR product, and the immobilization reaction temperature was 80 ° C. Due to the nature of the probe used, a thermal denaturation reaction is required. For this purpose, it is treated for 2 minutes in 100 ℃ distilled water.

As shown in Table 9, the specificity of the plastic substrate according to the present invention maintained a value equivalent to that of the plastic substrate manufactured by the conventional wet process.

TABLE 9

division Target H Target I Target J Plastic substrate of the present invention 3.3 2.3 3.0 Control 1.6 2.4 1.5

According to the method of the present invention, there is provided a plastic substrate which can significantly reduce the self-luminescence and thereby significantly improve the detection specificity of the target biomolecule. Therefore, in spite of the advantages of various designs including microstructures compared to organic substrates, plastic substrates, which have been limited in use due to the disadvantage of high self-luminescence, can be widely used for microarrays, biochips or well plates. In addition, according to the present invention it is possible to improve the specificity of the substrate surface and to easily adjust the characteristics.

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Claims (20)

In the manufacturing method of a plastic substrate used for a microarray, a biochip, or a well plate, a) molding a plastic substrate having an AFM (Atomic Force Microscope) surface roughness of R _a <3 nm or R _q <4 nm under conditions of 50 μm × 50 μm or less; b) treating the plastic substrate with a plasma; And c) treating the substrate with a monomer for surface modification comprising at least one functional group selected from the group consisting of an epoxide functional group, an aldehyde functional group and an amine functional group; Method for producing a plastic substrate excellent in detection specificity of the target biomolecule by including. The method of claim 1, Wherein said plasma treatment step and surface treatment monomer treatment step are performed simultaneously. The method of claim 1, And said plasma is an argon plasma. The method of claim 1, The plasma is a continuous plasma or pulsed plasma having an energy density of 10 ⁸ J / kg or less. The method of claim 4, wherein The plasma is treated for 10 minutes or less. The method of claim 1, The frequency of the plasma is microwave (2.45 GHz) or RF (13.56 MHz). The method of claim 1, The plastic substrate is a transparent resin selected from the group consisting of polystyrene (PS), cycloolefin copolymer (COC) and polycarbonate (PC), or a translucent resin selected from the group consisting of polypropylene (PP) and polyethylene (PE). Characterized in that the method. The method of claim 1, The surface modification monomer is an allyl glycidyl ether or glycidyl methacrylate containing an epoxide functional group and having a carbon double bond, whereby the plastic surface is an epoxide group. Modified method. The method of claim 1, The surface modification monomer is hexanal (Hexanal), octanal (Octanal) or nonanal (Nonalal) having a boiling point of at least 80 ℃ containing an aldehyde group functional group, whereby the plastic surface is characterized in that the modified aldehyde group. . The method of claim 1, The surface modification monomer is an allylamine or an ether compound, 3-methoxypropylamine, which contains a primary amine function and has a carbon double bond, whereby the plastic surface is modified with an amine group. Characterized in that the method. The method of claim 1, And forming a microfluidic structure on the plastic substrate. Atomic Force Microscope (AFM) surface roughness with R _a <3 nm or R _q <4 nm under conditions of 50 μm × 50 μm and selected from the group consisting of plasma and epoxide functional groups, aldehyde functional groups and amine functional groups on the surface A plastic substrate used in a microarray, a biochip, or a well plate, characterized in that the monomer for surface modification comprising one or more functional groups has been treated, and having excellent detection specificity of the target biomolecule. The method of claim 12, The plasma is a plastic substrate, characterized in that the argon plasma. The method of claim 12, The plasma is a plastic substrate, characterized in that the continuous plasma or pulsed plasma having an energy density of 10 ⁸ J / kg or less. The method of claim 14, And the plasma is processed for 10 minutes or less. The method of claim 12, The frequency of the plasma is a plastic substrate, characterized in that microwave (2.45 GHz) or RF (13.56 MHz). The method of claim 12, It is a transparent resin selected from the group consisting of polystyrene (PS), cycloolefin copolymer (COC) and polycarbonate (PC) or translucent resin selected from the group consisting of polypropylene (PP) and polyethylene (PE). Plastic substrate. The method of claim 12, The surface modification monomer is an allyl glycidyl ether or glycidyl methacrylate having an epoxide functional group and having a carbon double bond, whereby the surface is modified with an epoxide group. Plastic substrate, characterized in that. The method of claim 12, The surface modification monomer is a hexanal (Hexanal), octanal (Octanal) or nonanal (Nonalal) having a boiling point of more than 80 ℃ containing an aldehyde group functional group, whereby the surface is modified with an aldehyde group. The method of claim 12, The surface-modifying monomer is an allylamine or an ether compound, 3-methoxypropyl amine (3-Methoxypropylamine) containing a primary amine functional group, whereby the surface is modified with an amine group. Plastic substrate.

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