KR20110117282A - Manufacturing method for flexible biosensor and sensing method using the same - Google Patents

Abstract

Provided are a flexible biosensor using a laser and a detection method using the same.
Flexible biosensor according to the present invention is a flexible lower substrate;
A silicon substrate stacked on the flexible lower substrate and formed with source and drain regions doped with first type impurities separated by predetermined intervals; And a source, a drain, and a gate electrode stacked on the silicon substrate, wherein a detection material for detecting a biologically active material is immobilized on the gate electrode, and the silicon substrate is crystallized by a laser. Since the biosensor according to the present invention is directly implemented on the flexible substrate, the device is manufactured on a silicon substrate, and then, compared to the conventional technology of transferring the device substrate to the flexible substrate, the economy is excellent.

Description

Flexible biosensor using laser and detection method using same {Manufacturing method for flexible biosensor and sensing method using the same}

The present invention relates to a method for manufacturing a flexible biosensor using a laser, a flexible biosensor manufactured by the same, and a detection method using the same. More specifically, the present invention can effectively overcome the limitations of a substrate having a biosensor implemented on a conventional silicon substrate. The present invention relates to a method of manufacturing a flexible biosensor using an economical laser, a flexible biosensor manufactured by the same, and a detection method using the same.

Organisms, including humans, have many sensory organs that sense not only the five senses but also various stimuli from the outside, such as pain or, of course, pain or temperature sensing. This sensed stimulus is compared to the stimulus data already educated by experience in the brain to recognize subtle tastes and flavor changes. These functions are called sensory organs in living things, and sensors in machines and instruments. Biosensors that can detect physicochemical stimuli from the outside by applying electronics to simulate biological functions are commonly called biosensors.

However, conventional biosensors are difficult to have various structures because they are fabricated on a microarray or microfluidic channel (hereinafter referred to as a microfluidic channel) formed on a rigid substrate such as a silicon substrate. In order to overcome the limitations of such a substrate, Lieber et al. Disclose a so-called bottom-up sensing device manufacturing method for growing silicon nanowires using a catalyst on a substrate. However, since the bottom-up sensing device must grow nanowires directly on the substrate, there is a problem that the performance of the overall semiconductor device is degraded and the homogeneity is poor (Nature Biotechnology vol. 23, 1294, 2005).

In order to solve the disadvantage of the bottom-up sensing device manufacturing method, McAlpine et al. Is a top-down method using a microstructure semiconductor (μ-Sc) technology, nanowires on a plastic substrate Disclosed is a chemical sensor using (Nature Materials, Vol 6, May 2007, hereinafter prior art). However, the prior art relates to a technique for detecting a certain component of a gas, which is difficult to apply directly to a biosensor made of a solvent such as water, and if a plurality of kinds of substances are to be detected, a plurality of sensors may be manufactured. There is a problem that should be.

Accordingly, a new concept of a flexible high-sensitivity biosensor, particularly a semiconductor sensor, implemented on a flexible substrate and capable of sensing a plurality of types of detection materials very effectively using a high performance semiconductor has not yet been implemented. In particular, the extreme conditions of the semiconductor manufacturing process and the weak heat resistance and chemical characteristics of the flexible substrate (generally polymer material) and the biomaterial are considered to be incompatible, and as a result, the biosensors, particularly semiconductors, implemented on the flexible substrate Biosensor using the situation is not yet disclosed.

Furthermore, biosensors using various metals require chemical pretreatment processes to immobilize biologically active materials (proteins, peptides) on chip electrodes. However, the method has a big disadvantage such as complex and non-specific protein binding, and because it is not only a weak binding force, but also can be affected by many chemicals, there are many limitations when testing various protein-protein interactions, making it difficult to apply. . Furthermore, when this chemical process is performed on a flexible substrate such as plastic, it may adversely affect the substrate itself.

Accordingly, the problem to be solved by the present invention is to detect the effective bio-materials according to the specific binding between biologically active substances, and due to the flexibility of the substrate, a new concept flexible biosensor and a detection method using the same To provide.

Another problem to be solved by the present invention is effective bio-material detection according to the specific binding of the bio-active material, and due to the flexibility of the substrate effective and economical of a new concept flexible biosensor with high applicability It is to provide a manufacturing method.

In order to solve the above problems, the present invention is a flexible lower substrate; A silicon substrate stacked on the flexible lower substrate and formed with source and drain regions doped with first type impurities separated by predetermined intervals; And a source, a drain, and a gate electrode stacked on the silicon substrate, wherein a detection material for detecting a biologically active material is immobilized on the gate electrode, and the silicon substrate is crystallized by a laser. Provide a sensor.

The present invention also provides a flexible lower substrate; A silicon upper substrate in contact with an upper portion of the flexible lower substrate, the source and drain regions being spaced apart at predetermined intervals; A microfluidic channel passing through the silicon substrate between the source and drain regions, wherein a detection material for detecting a biologically active material is immobilized on the silicon substrate between the source and drain regions, wherein the silicon substrate is Provided is a flexible biosensor characterized in that it is crystallized. The source and drain regions are stacked on the silicon substrate, and are formed by irradiating the doped layer doped with the first type impurities with the laser to diffuse the first type impurities into the silicon substrate. It includes, the detection material may be a fusion protein in the form of a gold binding protein and the detection protein fused. In this case, the flexible biosensor further includes a microfluidic channel passing through the gate electrode, and the detection material may be immobilized to the gate electrode in such a manner as to flow the detection material through the microfluidic channel. The laser is an excimer laser, and the first type impurity is an n type impurity. A detection method using the above-described biosensor, the method comprising: flowing a target material into a microfluidic channel passing through a gate electrode on a silicon substrate; And measuring a change in current of the biosensor according to the coupling between the target material and the detection material. Alternatively, the detection method using a biosensor according to another embodiment of the present invention, the method includes flowing a target material in the microfluidic channel between the source and drain regions; And measuring a change in current of the biosensor according to the binding between the target material and the detection material. The microfluidic channel may simultaneously pass through one or more gate electrodes.

In order to solve the above another problem, the present invention comprises the steps of laminating an amorphous first silicon layer on a flexible substrate; Stacking a doped layer doped with a first type impurity on the amorphous first silicon layer; Patterning the doped layer to form source and drain region doped layers spaced at predetermined intervals; The first silicon layer and the source and drain region doped layers are irradiated with a laser to crystallize the first silicon layer, and at the same time, the impurities of the n-type doped layer are diffused into the first silicon layer below. Forming source and drain regions in the trenches; Patterning the first silicon layer to form a silicon device substrate including the source and drain regions; Stacking and patterning a gate oxide layer on the device substrate to expose the source and drain regions; Stacking and patterning a metal layer on the gate oxide layer to fabricate source, gate, and drain electrodes; And forming a microfluidic channel passing through the gate electrode pad extending from the gate electrode.

The invention also includes forming a bottom gate electrode on the flexible substrate; Stacking an insulating layer on the lower gate electrode and the flexible substrate; Depositing an amorphous first silicon layer on the insulating layer; Stacking a doped layer doped with a first type impurity on the amorphous first silicon layer; Patterning the doped layer to form source and drain region doped layers spaced at predetermined intervals; The first silicon layer and the source and drain region doped layers are irradiated with a laser to crystallize the first silicon layer, and at the same time, the impurities of the n-type doped layer are diffused into the first silicon layer below. Forming source and drain regions in the trenches; Forming source and drain electrodes on the source and drain regions; And fabricating a microfluidic channel passing through the silicon substrate between the source and drain regions.

The biosensor manufacturing method according to an embodiment of the present invention flows the biologically active material into the microfluidic channel passing through the gate electrode pad through a biologically active material that can be specifically bound to the gate electrode. It may further comprise the step of fixing to.

Biosensor manufacturing method according to another embodiment of the present invention by flowing a biologically active material capable of specifically binding to a silicon substrate to the microfluidic channel passing through the silicon substrate between the source and drain regions, the biological activity The method may further include fixing a material to the silicon substrate. The first type impurity may be an n type impurity, and the preparing of the microfluidic channel may be performed on the silicon substrate, the source and drain electrodes, and the passivation layer exposing the silicon substrate between the source and drain electrodes. ; And stacking an upper cover layer laminated on the passivation layer and bonded to the passivation layer. In addition, the upper cover layer may be provided with a hole for injecting or discharging a sample into the microfluidic channel.

Since the biosensor according to the present invention is directly implemented on the flexible substrate, the biosensor is manufactured in a silicon substrate, and then, compared to the conventional technology of manufacturing a device substrate and transferring the flexible substrate to the flexible substrate, the economic efficiency is excellent. In addition, since the biosensor device according to the present invention detects a biomaterial directly from a plastic substrate by using a high performance microstructure semiconductor, the biosensor device has a superior sensitivity than a conventional biosensor. Furthermore, in order to overcome the limitations of the flexible substrate, the biosensor according to the present invention can specifically bind the desired biologically active material to the electrode pad without a separate pretreatment of the metal electrode pad, and thus, economical and process applicability great. That is, compared to the conventional self-assembled monolayer (SAM) -based biomaterial immobilization technology, the surface of the bioreceptor can be efficiently functionalized with the desired bioreceptor in a simpler process while maintaining the orientation of the bioreceptor without the surface modification process. Can be. In addition, when a high-sensitivity biosensor based on electrical detection is implemented on a transparent plastic substrate, the bio signal is converted into a digital electrical signal, thereby increasing the linkage with other information processing devices, and being portable and providing electrical detection as well as optical detection. There are many benefits, including lower costs.

1 to 12 are steps illustrating a method of manufacturing a biosensor according to an embodiment of the present invention.
13 to 16 are diagrams showing an example of protein detection using a biosensor according to an embodiment of the present invention.
17 and 18 are diagrams showing examples of DNA detection using a biosensor according to an embodiment of the present invention.
19 to 30 are steps illustrating a method of manufacturing a biosensor using a silicon bonding material.
31 to 34 are diagrams showing examples of protein detection using a biosensor according to an embodiment of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the drawings. The following embodiments are provided as examples to ensure that the spirit of the present invention to those skilled in the art will fully convey. Therefore, the present invention is not limited to the embodiments described below, but may be embodied in other forms. In the drawings, the width, length, thickness, etc. of the components may be exaggerated for convenience. Like reference numerals denote like elements throughout the specification, and the accompanying drawings are all interpreted in the form of a cross-sectional view of the entire plan view and the partial cross section A-A '.

The present invention manufactures a silicon-based semiconductor device implemented in a flexible substrate in such a way that the amorphous silicon substrate previously laminated on the flexible substrate is processed by laser to crystallize. In other words, the device substrate of the biosensor is defined by forming a source and a drain, and then a new method of manufacturing a semiconductor device-based biosensor directly from the flexible substrate using a laser, rather than transferring the corresponding region to the flexible substrate. It provides a biosensor manufacturing method. As used herein, the term “flexible” is a term that is distinguished from a silicon substrate having a rigid characteristic and the like, and may be bent or folded at a predetermined angle, such as a plastic substrate or a thin film silicon substrate. A term that includes all of the properties present.

As described above, the present invention manufactures a biosensor for detecting a specific biomaterial by a laser based method. In particular, the present invention is economical compared to the prior art, since a biosensor of a semiconductor device can be manufactured directly from a silicon substrate provided in a plastic substrate, unlike the prior art in which a device substrate is manufactured on a silicon substrate and then transferred to PDMS. . The biosensor manufacturing method according to the present invention can be applied to any biosensor based on a crystallized silicon substrate, but in one embodiment (Example 1) in which a detection material is immobilized in a metal gate electrode of the present invention and a detection material on a silicon substrate This provides a fixed form of work.

Hereinafter, a method of manufacturing a biosensor according to the present invention will be described in detail with reference to the drawings.

Example  One

Biosensor manufacturing using gold binding material

1 to 12 are steps illustrating a method of manufacturing a biosensor according to an embodiment of the present invention.

Referring to FIG. 1, a flexible substrate, such as a plastic substrate 100, is disclosed, rather than a rigid substrate such as a silicon substrate. That is, the present invention manufactures biosensors directly from a flexible substrate such as plastic, unlike the prior art, which manufactures all or part of biosensors on silicon substrates.

Referring to FIG. 2, a silicon oxide layer, for example, a silicon oxide layer 110 by PECVD is stacked on the flexible substrate 100 to a predetermined thickness, and the oxide layer 110 functions as a kind of buffer layer.

Referring to FIG. 3, a first silicon layer (amorphous silicon, a-Si, 120), which is amorphous silicon, is stacked on the oxide layer 110. In one embodiment of the present invention, the amorphous first silicon layer was deposited by PECVD.

Referring to FIG. 4, a silicon doped layer 130 doped with a first type impurity is stacked on the amorphous first silicon layer 120. In one embodiment of the present invention, the first type impurity is an n type doping layer doped with n type impurities, for example, PH3. The impurity doping method may be a method such as an ion implant, but the scope of the present invention is not limited thereto. In one embodiment of the present invention, a phosphine gas was simultaneously flowed while the amorphous silicon layer was deposited on the first silicon layer 120 by PECVD. Thus, the phosphine doped layer 130 is formed on the first silicon layer 120.

Referring to FIG. 5, the doped layer 130 is selectively etched to leave only the doped layers 130a and 130b corresponding to the source and drain regions on the amorphous silicon layer 120. In an embodiment of the present invention, the doped layer is patterned by a photolithography process, and then the remaining regions not protected by the mask are removed by a wet etching process.

Referring to FIG. 6, the amorphous first silicon layer 120 is crystallized using a laser. The excimer laser used in the crystallization of amorphous silicon in the present invention encapsulates two mixed gases in a vacuum vessel, and then stimulates with a discharge or an electron beam to generate an unstable excited dimer, which is released at high power as they collapse. Refers to an ultraviolet beam. The excimer laser has the advantage of being extremely high in compatibility with semiconductor materials because the excimer laser generates only a large output evenly, has a small diffraction phenomenon, and is chemically operated without thermal phenomenon when interacting with the material. In particular, heat treatment and crystallization by laser instead of conventional RTP (Rapid Thermal Processing) has the advantage of having a thermal burden close to '0'. That is, since the laser pulse is irradiated for about 108 times shorter than the RTP (nanosecond unit), it is possible to crystallize even if the thermal barrier layer is required even on the plastic substrate. To manufacture a biosensor based. The laser treatment according to the invention has the advantage of being heat treatable in local and selective areas. That is, laser technology can reduce the inefficient crystallization process in the undesired region because the high temperature heat treatment is possible in the local and selective region by using the beam when compared to the RTP that the entire wafer is heat-treated under the same conditions.

The present invention irradiates an excimer laser directly onto a silicon substrate stacked on a flexible substrate, whereby the first impurity (n-type) of the doped layer in which the source and drain regions are formed, as well as the crystallization of the amorphous first silicon layer 120 is formed. Impurities) are diffused into the amorphous silicon layer 120 below, so that the source and drain regions S and D are formed in the first silicon layer 120a. The doped layer of the first type impurity is subsequently removed by photolithography and dry etching, and thus the n-type impurity is doped in the crystallized first silicon layer 120a and a predetermined region of the first silicon layer 120a. Source and drain regions are formed.

Referring to FIG. 7, the crystallized first silicon layer 120a is patterned to form a device substrate (hereinafter referred to as a bio device substrate 120b) of a biosensor transistor including a source and a drain region.

Referring to FIG. 8, an insulating layer 140 such as silicon oxide is stacked on the bio device substrate 120b. Stacking of the insulating layer 140 is performed in the same manner as chemical vapor deposition, and the insulating layer 140 functions as a gate oxide film in the silicon substrate between the source and drain (S, D).

Referring to FIG. 9, the insulating layer 140 is patterned, the source and drain (S, D) regions of the bio device substrate 120b are exposed, and the insulating layer 140 is still stacked in the gate region therebetween. Maintain state. The insulating layer 140 on the gate region then functions as a gate oxide.

Referring to FIG. 10, a metal layer 150 is stacked on the insulating layer 140, thereby forming source and drain electrodes in direct contact with the source and drain regions. The gate electrode is stacked on the bioelement substrate with the insulating layer interposed therebetween.

Referring to FIG. 11, the metal layer 150 is patterned to form source, drain electrodes, and gate electrodes 150a, 150c, and 150b. In addition, the source, drain, and gate electrodes extend to a predetermined length, and pads having a wider width are provided at the extended electrode ends. In particular, the gate electrode 150b is provided in a substrate region spaced apart from the device substrate region, and a microfluidic channel through which a biologically active material flows is formed in the gate electrode pad region.

In order to make the microfluidic channel, the present invention uses a PDMS having trenches of a predetermined depth, which is disclosed in FIG. Referring to FIG. 12, the PDMS 200 having the trench 210 having a predetermined depth is brought into contact with the gate electrode pad 120b so that the trench 210 passes through the gate electrode pad 150b. The fluid channel 210 is formed. In one embodiment of the present invention, the electrode material was gold, and a polymer layer 220 such as SU-8 was deposited between the PDMS 200 and the silicon oxide layer 110 for sealing between the PDMS and the flexible substrate. Can be. As a result, a biosensor in the form of a flexible transistor capable of detecting a current change according to a voltage change according to the reaction of the biomaterial in the gate electrode pad is manufactured.

Hereinafter, a detection example of a biologically active substance of the flexible biosensor according to an embodiment of the present invention will be described in detail.

Experimental Example  One

Protein Detection Using Gold Binders

Experimental Example  1-1

Antigen immobilization

In the present embodiment, a fusion protein (hereinafter referred to as GBP-fusion protein) in which a gold binding protein (GBP) and a desired target protein are fused is flowed into a microfluidic channel, thereby allowing the fusion protein to flow through the gate electrode pad 150b. Immobilized on.

13 to 16 are diagrams showing an example of protein detection using a biosensor according to an embodiment of the present invention.

Referring to FIG. 13, the GBP-fusion protein flows into the microfluidic channel 210 of the biosensor according to the present invention, and the flowing GBP-fusion protein is specifically bound to and immobilized on the gate electrode pad 150b. In the present embodiment, the target protein is Avian Influenza Viral Surface Antigen (Korea specific H5N1 & H9N2 AIa), and is fused with GBP. GBP used in this example follows the following amino acid sequence.

1. GBP1

H ₂ N- MHGKTQATSGTIQS -COOH

2. GBP3

H ₂ N- MGKTQATSGTIQSMHGKTQATSGTIQSMHGKTQATSGTIQS-COOH

3. GBP10

H ₂ N- SKTSLGQSGASLQGSEKLTNG-COOH

Referring to FIG. 14, it can be seen that the GBP-antigen fusion protein (GBP-AIa) is combined and fixed to the gold electrode pad 150b. As a result, a change in current according to a change in voltage of the gate electrode is detected (see the graph above).

Experimental Example  1-2

Antibody detection

Referring to FIG. 15, the same or another microfluidic channel 310 is disclosed passing through the gate electrode pad 150b to which the GBP-antigen fusion protein is coupled. The target material containing the antibody is flowed through the microfluidic channel 150b, and if there is an antibody that specifically binds the antigen in the target material, specific binding between the antigen and the antibody occurs, thereby causing a gate electrode. The voltage at 150b is changed. As described above, the microfluidic channel 310 may be formed by contacting a trench having a predetermined depth and width to face the gate electrode pad 150b.

In one embodiment of the invention, the target material to be detected flows through another microfluidic channel that passes through all of the plurality of gate electrode pads (A, B, C), whereby multiple kinds of antigens for the same antibody Can be detected. However, the scope of the present invention is not limited thereto.

16 is a schematic diagram of antigen-antibody binding and current change graphs according to antigen-antibody binding for the present example. In this embodiment, the antibody is an avian influenza antibody that specifically binds to an AIa antigen. Referring to FIG. 16, it can be seen that the gate voltage change and the current change are detected at a remarkable level according to the antigen-antibody binding.

Hereinafter, a detection example of a biologically active substance of the flexible biosensor according to an embodiment of the present invention will be described in detail.

Experimental Example  2

DNA  detection

The biosensor according to the present invention is capable of detecting DNA as well as protein, in which case the DNA is detected according to specific hybridization between the target DNA and the detection DNA.

17 and 18 are diagrams showing an example of DNA detection using a biosensor according to another embodiment of the present invention, and FIG. 17 is a schematic diagram of DNA detection according to an embodiment of the present invention.

Referring to FIG. 17, a single stranded DNA containing a thiol group (SH) at its end is bonded to a gate electrode pad (gold electrode pad) according to the bond between the thiol group (-SH) and the gold electrode pad. As a result, the single stranded DNA containing the thiol group as an end group is fixed to the gate electrode pad as the detection DNA (Probe DNA). The target DNA is then flowed into the microfluidic channel. When the target DNA has a nucleotide sequence complementary to the detection DNA, hybridization occurs between the target DNA and the detection DNA. The voltage of the gate electrode is changed according to the charge change due to the hybridization bond, and thus the current change is detected.

18 is a schematic diagram of a current change according to DNA hybridization according to an embodiment of the present invention.

Referring to FIG. 18, it can be seen that a current change occurs as the detection DNA containing a thiol group as an end group is fixed to the gate electrode pad (top), and a current change occurs as a result of hybridization with the target DNA (bottom). .

Example  2

Biosensor Manufacturing Using Silicon Bond Material

19 to 30 is a view showing a method of manufacturing a biosensor using a silicon bonding material.

Referring to FIG. 19, a flexible substrate 800 such as plastic is provided.

Referring to FIG. 20, a lower gate electrode 810 is stacked on the plastic substrate 800.

In one embodiment of the present invention, the lower gate electrode 810 is made of a metal made of chromium (Cr) and gold (Au), but the scope of the present invention is not limited thereto.

Referring to FIG. 21, a gate insulating layer 820 is stacked on the lower gate electrode 810 and the plastic substrate 800 at a predetermined height. As a result, the gate electrode 810 maintains an electrical insulation state with the upper element. Silicon oxide (SiO ₂ ) may be used as the gate insulating layer 820, and may be deposited by plasma chemical vapor deposition (PECVD).

Referring to FIG. 22, an amorphous first silicon layer 830 is stacked on the insulating layer 820, and the amorphous first silicon layer may be performed by a PECVD process or the like.

Referring to FIG. 23, a doping layer 840 doped with n-type impurities as first type impurities is stacked on the first silicon layer 830. Since the doping layer 840 is stacked in the same manner as in the first embodiment, it will be omitted below.

Referring to FIG. 24, the doped layer 840 is patterned to match source and drain regions spaced at predetermined intervals. Here, the source and drain regions mean a substrate region in which the source and drain electrodes are formed in the transistor, and in the present invention, in particular, the source and drain regions of the transistor are formed by diffusing the silicon substrate under the impurities of the doping layer 840. As described above, the diffusion may be performed by laser treatment.

Referring to FIG. 25, the amorphous first silicon layer 830 and the patterned doped layer 840 are laser treated, whereby the first type impurity of the doped layer with the crystallization of the amorphous silicon is deposited below the first silicon layer. As it diffuses into, the source and drain regions S and D of the silicon substrate are formed. Thus, the crystallized first silicon layer 830a and the source and drain regions S and D formed in the first silicon layer 830a are manufactured. As described above, the present invention performs the manufacture of a semiconductor device directly on a plastic substrate through a laser without a separate transfer process.

Referring to FIG. 26, the crystallized first silicon layer 830a is patterned to form a transistor substrate of a biosensor including a source and a drain region.

Referring to FIG. 27, source and drain electrodes 840 and 850 are stacked in the source and drain regions S and D of the first silicon layer 830a. This completes a transistor element consisting of a lower gate electrode, an upper source and a drain electrode.

28 to 29, in order to form a microfluidic channel in a gate region of a silicon substrate of the transistor device, a passivation layer 900 made of a material such as SU-8 is first stacked on a silicon substrate. In this case, the passivation layer 900 has a trench structure in which only a gate substrate is exposed (see FIG. 28). Thereafter, a cover layer 910 made of a flexible material such as PDMS is stacked on the passivation layer 900 to form a microfluidic channel passing only through the gate substrate. In particular, by stacking a passivation layer such as SU-8 on the silicon substrate first, it is possible to prevent the leakage of the sample from the microfluidic channel, and the trench and the sample facing the trench of the passivation layer 900 in the cover layer 910 itself. By providing a hole capable of injecting and discharging, the sample fluid can be easily flowed and discharged. The present invention couples the detection material to the gate substrate by flowing a biologically active material that specifically binds to the gate region through the microfluidic channel passing through the gate region of the silicon substrate. To this end, the present invention provides a fusion protein in which a silicon-binding protein that specifically binds to silicon and a target material are fused, which will be described in more detail below.

30 is a graph showing the transistor effect of the biosensor according to the present invention shown in FIG.

Referring to FIG. 30, it can be seen that the collector current increases as the base voltage increases. Thus, the results show that the biosensors produced on plastic substrates according to the present invention exhibit typical transistor characteristics.

Hereinafter, a method for implementing a biosensor manufactured according to the present invention will be described in detail with reference to the accompanying drawings.

Experimental Example  3

Antigen detection

The detection protein was fixed by flowing a protein fused with a protein (Silica Binding Protein) and a detection protein selectively bound to a silicon substrate through the microfluidic channel of the biosensor shown in FIG. 31.

SBP used in this experimental example follows the following nucleotide sequence and amino acid sequence.

1. rplB1

5'-GCTATCGTTAAATGTAAGCCGACCTCCGCTGGTCGTCGTCACGTTGTTAAAATCGTGAACCCTGAATTACATAAGGGTAAACCTTACGCACCTTTATTAGATACTAAATCTAAAACTGGTGGTCGTAATAATTTAGGACGTATCACTACTCGTCATATCGGTGGTGGTCATAAACAA -3 ’

RplB1

H ₂ N-AIVKCKPTSAGRRHVVKIVNPELHKGKPYAPLLDTKSKTGGRNNLGRITTRHIGGGHKQ-COOH

2. rplB2

5'-GTACTTGGTAAAGCCGGTGCCAACCGCTGGAGAGGCGTTCGCCCTACAGTTCGCGGTACTGCGATGAACCCGGTAGATCACCCGCACGGTGGTGGTGAAGGTCGTAACTTTGGTAAACACCCGGTATCACCTTGGGGCGTTCAAACCAAAGGTAAGAAAACTCGTCATCACAAATA

RplB2

H ₂ N-VLGKAGANRWRGVRPTVRGTAMNPVDHPHGGGEGRNFGKHPVSPWGVQTKGKKTRHNKRTDKYIVRRRGK-COOH

3. rplB12

5'-ATGGCTATCGTTAAATGTAAGCCGACCTCCGCTGGTCGTCGTCACGTTGTTAAAATCGTGAACCCTGAATTACATAAGGGTAAACCTTACGCACCTTTATTAGATACTAAATCTAAAACTGGTGGTCGTAATAATTTAGGACGTATCACTACTCGTCATATCGGTGGTGGTCATAAACAAgtcgacGTACTTGGTAAAGCCGGTGCCAACCGCTGGAGAGGCGTTCGCCCTACAGTTCGCGGTACTGCGATGAACCCGGTAGATCACCCGCACGGTGGTGGTGAAGGTCGTAACTTTGGTAAACACCCGGTATCACCTTGGGGCGTTCAAACCAAAGGTAAGAAAACTCGTCACAACAAACGTACCGATAAATATATCGTACGTCGTCGTGGCAAA-3 '

RplB12

H ₂ N-MAIVKCKPTSAGRRHVVKIVNPELHKGKPYAPLLDTKSKTGGRNNLGRITTRHIGGGHKQVDVLGKAGANRWRGVRPTVRGTAMNPVDHPHGGGEGRNFGKHPVSPWGVQTKGKKTRHNKRTDKYIVRRRGK-COOH

In the present invention, SBP of the following base sequence and amino acid sequence is used.

SBP1-coding gene

5'-ATGAGCCCACACCCGCACCCACGTCACCATCACACC-3 '

SBP1

H ₂ N-MSPHPHPRHHHT-COOH

SBP5-coding gene

5'-AAACCGAGCCACCACCACCACCACACCGGCGCGAAC-3 '

SBP5

H ₂ N- KPSHHHHHTGAN-COOH

SBP10-coding gene

5'-CGTGGCCGTCGTCGTCGTCTGTCTTGCCGTCTGCTG-3 '

SBP10

H ₂ N-RGRRRRLSCRLL-COOH

In the present experimental example, PBS was flowed into the microfluidic channel and washed first, followed by fusion protein of SBP-antigen (SBP-AIa). The antigen used in one embodiment of the present invention is H5N1 & H9N2 Avian influenza viral surface antigens, and has the sequence of H ₂ N-CRDNWKGSNRPI-COOH. In the method for producing SBP-antigen fusion protein (SBP-AIa), a recombinant vector configured to be expressed in a fused form of two genes is inserted into E. coli, transformed, and cultured the transformed microorganism, It was a way to express the fusion protein (SBP-AIa). The fusion protein specifically binds to the gate region of the silicon substrate by SBP.

Referring to Figure 32, it can be seen that the current change of the biosensor according to the present invention is detected according to the binding of the antigen (see below in the figure).

Referring to FIG. 33, an antibody is shed in a silicon substrate region (gate region) of the biosensor to which an antigen is bound, thereby causing another specific binding of the antibody and the antibody specifically bound to silicon.

Referring to Figure 34, it can be seen that the change of the collector current of the biosensor occurs according to the antigen-antibody binding.

Claims (17)

A flexible lower substrate;
A silicon substrate stacked on the flexible lower substrate and formed with source and drain regions doped with first type impurities separated by predetermined intervals; And
A source, a drain, and a gate electrode stacked on the silicon substrate, wherein a detection material for detecting a biologically active material is immobilized on the gate electrode, and the silicon substrate is crystallized by a laser. . A flexible lower substrate;
A silicon upper substrate in contact with an upper portion of the flexible lower substrate, the source and drain regions being spaced apart at predetermined intervals;
A microfluidic channel passing through the silicon substrate between the source and drain regions, wherein a detection material for detecting a biologically active material is immobilized on the silicon substrate between the source and drain regions, wherein the silicon substrate is Flexible biosensor, characterized in that the crystallized. 3. The method according to claim 1 or 2,
The source and drain regions,
A flexible biosensor, stacked on the silicon substrate, formed by dispersing a doping layer doped with a first type impurity with the laser to diffuse the first type impurity onto the silicon substrate. The method of claim 1,
The gate electrode includes gold, and the detection material is a flexible biosensor, characterized in that the fusion protein in the form of a fusion of the gold binding protein and the detection protein. The method of claim 1,
The flexible biosensor further comprises a microfluidic channel passing through the gate electrode, wherein the detection material is immobilized on the gate electrode in such a manner as to flow the detection material through the microfluidic channel. 3. The method according to claim 1 or 2,
The laser is excimer laser, characterized in that the biosensor manufacturing method using a laser. 3. The method according to claim 1 or 2,
And said first type impurity is an n-type impurity. A detection method using a biosensor according to claim 1, comprising: flowing a target material into a microfluidic channel passing through a gate electrode on a silicon substrate; And
And measuring a change in current of the biosensor according to the coupling between the target material and the detection material. A detection method using a biosensor according to claim 2, the method comprising: flowing a target material into a microfluidic channel between the source and drain regions; And
And measuring a change in current of the biosensor according to the coupling between the target material and the detection material. The method of claim 8,
The microfluidic channel is characterized in that passing through at least one gate electrode at the same time, the detection method using a biosensor. Depositing an amorphous first silicon layer on the flexible substrate;
Stacking a doped layer doped with a first type impurity on the amorphous first silicon layer;
Patterning the doped layer to form source and drain region doped layers spaced at predetermined intervals;
The first silicon layer and the source and drain region doped layers are irradiated with a laser to crystallize the first silicon layer, and at the same time, the impurities of the n-type doped layer are diffused into the first silicon layer below. Forming source and drain regions in the trenches;
Patterning the first silicon layer to form a silicon device substrate including the source and drain regions;
Stacking and patterning a gate oxide layer on the device substrate to expose the source and drain regions;
Stacking and patterning a metal layer on the gate oxide layer to fabricate source, gate, and drain electrodes; And
And forming a microfluidic channel passing through the gate electrode pad extending from the gate electrode. Forming a lower gate electrode on the flexible substrate;
Stacking an insulating layer on the lower gate electrode and the flexible substrate;
Depositing an amorphous first silicon layer on the insulating layer;
Stacking a doped layer doped with a first type impurity on the amorphous first silicon layer;
Patterning the doped layer to form source and drain region doped layers spaced at predetermined intervals;
The first silicon layer and the source and drain region doped layers are irradiated with a laser to crystallize the first silicon layer, and at the same time, the impurities of the n-type doped layer are diffused into the first silicon layer below. Forming source and drain regions in the trenches;
Forming source and drain electrodes on the source and drain regions; And
And manufacturing a microfluidic channel passing through the silicon substrate between the source and drain regions. 12. The method of claim 11,
And securing the biologically active material to the gate electrode pad by flowing a biologically active material capable of specifically binding to the gate electrode to the microfluidic channel passing through the gate electrode pad. Biosensor manufacturing method using a laser. The method of claim 12,
Securing the biologically active material to the silicon substrate by flowing a biologically active material capable of specifically binding to the silicon substrate into a microfluidic channel passing through the silicon substrate between the source and drain regions. A biosensor manufacturing method using a laser. The method of claim 11 or 12,
The first type impurity is a biosensor manufacturing method, characterized in that the n-type impurity. The method of claim 12,
The manufacturing of the microfluidic channel may include stacking a passivation layer on the silicon substrate, the source and drain electrodes, and exposing the silicon substrate between the source and drain electrodes; And
Stacking on the passivation layer, comprising the step of laminating an upper cover layer bonded to the passivation layer, biosensor manufacturing method using a laser. 17. The method of claim 16,
The upper cover layer is characterized in that a hole for injecting or discharging a sample into the microfluidic channel, characterized in that the biosensor manufacturing method using a laser.

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