KR100556580B1 - Carbon Nanotube Film with High Surface Density - Google Patents

Abstract

The present invention relates to a high-density CNT film obtained by laminating carbon nanotubes (CNT) by chemical bonding on a substrate exposed to an amine group.

According to the present invention, a CNT-biochip is formed by modifying a carboxyl group exposed to a CNT film fixed at a high density to a substrate with various chemical functional groups, and then chemically or physicochemically combining various bioreceptors having functional groups that bind to the chemical functional groups. It is possible to produce.

CNT, film, receptor, biochip, carboxyl group, modified

Description

Carbon Nanotube Film with High Surface Density}

1 is a schematic diagram of manufacturing a high-density CNT film by laminating a carboxyl group-exposed CNT with an amide bond on an amine-based substrate.

FIG. 2 shows that a CNT-DNA chip was prepared by binding DNA having an amino group to a CNT film exposed to a carboxyl group, and then performing a DNA hybridization reaction using the same.

Figure 3 shows that the CNT film exposed to the carboxyl group is modified with an amine group, and then a DNA having a carboxyl end group is bonded to prepare a CNT-DNA chip, and DNA hybridization is performed using the same.

4 is an XPS spectrum of phosphorous of the surface of a CNT film to which DNA is chemically bound.

FIG. 5 (a) shows a fluorescence image of a high density CNT film before binding DNA, and FIG. 5 (b) shows a fluorescence detection result of hybridization after binding DNA having amino terminal groups on the CNT film. (1) is a case of hybridization with complementary DNA, and (2) is a case of hybridization of non-complementary DNA.

FIG. 6 (a) shows fluorescence detection results when the DNA having the carboxyl end group is coupled to the amine group-modified CNT film, followed by hybridization of complementary DNA, and (b) hybridization of non-complementary DNA. The result of fluorescence detection in the case of reaction is shown.

The present invention relates to a high density carbon nanotube (CNT) film exposed to a chemical functional group on its surface.

CNT is a carbon allotrope composed of carbon present on the earth in large quantity. It is a material in which one carbon is combined with other carbon atoms in hexagonal honeycomb pattern to form a tube, and the diameter of the tube is nanometer (nm = 1 billionth of a meter). ) Is an extremely small area of matter. CNT is known as a perfect new material that has excellent mechanical properties, electrical selectivity, excellent field emission characteristics, high efficiency hydrogen storage medium properties and few defects among existing materials.

CNTs have excellent mechanical robustness and chemical stability, can exhibit both semiconductor and conductor properties, and because of their small diameter and relatively long length, CNTs are excellent materials for flat panel display devices, transistors and energy storage materials. In addition, the nanoscale application is very useful as various electronic devices.

In order to apply the above-described characteristics more diversely, single-wall CNTs were started to be chopped using strong acids. The CNTs thus cut have mainly -COOH functional groups at the cut end and part of the sidewall. These chemical functional groups have been used to chemically attach various materials to modify the properties of CNTs.

Furthermore, by replacing the functional group of CNT with -SH functional group by chemical mechanism, and then patterned on the surface by using micro contact printing technique on the gold surface (Nan, X. et al. , J. Colloid Interface Sci., 245: 311-8, 2002) and electrostatic methods have been used to adhere CNTs to surfaces using multiple films (Rouse, JH et al. , Nano Lett., 3: 59-62, 2003). However, the former has the disadvantage that the surface density of CNTs is low and the bonding strength is weak, and the latter has the fatal disadvantage that the patterning method of selectively fixing to the surface cannot be applied. Therefore, a new type of surface fixation method is urgently needed.

Of the 100,000 human genes currently estimated, about 10,000 functions have been identified, and most of these genes are known to be directly related to disease. In addition, since most diseases are caused at the protein level, not at the genetic level, more than 95% of the drugs developed or under development target proteins. Therefore, the study of biomolecules interacting with specific proteins and ligands, and based on the data obtained through protein function analysis and network analysis, to develop the treatment and prevention methods for diseases that were not possible with classical methods. Essential is an efficient protein-protein and protein-ligand reaction detection technique.

Recently, studies on reaction detection through electrochemical changes of CNTs immobilized with biomaterials have been made using the electrical, semiconductor properties, or structurally stable properties of CNTs (Dai, H. et al. , ACC. Chem). Res., 35: 1035-44, 2002; Sotiropoulou, S. et al. , Anal.Bioanal.Chem., 375: 103-5, 2003; Erlanger, BF et al. , Nano Lett., 1: 465-. 7, 2001; Azamian, BR et al. , JACS, 124: 12664-5, 2002).

A representative example of the protein-ligand reaction is the avidin-biotin reaction, in which a channel is formed on the substrate treated with polymer using CNTs, followed by electrochemical method for streptoavidin. Binding was measured (Star, A. et al. , Nano Lett., 3: 459-63, 2003).

CNTs are attracting attention as biochips because, first, they do not require labeling, second, they are very sensitive to signal changes, and third, they can proceed in aqueous solution without modification of proteins. The combination of new nanomaterials and biological systems will create important applications in disease diagnosis, genetics, and nanobiotechnology.

In order to develop faster and cheaper biochips, much research has recently been made on DNA hybridization detection technology. Various labeling techniques have been developed to detect hybridization of DNA, and fluorescent materials are most commonly used for labeling. By fixing a single DNA chain that can detect complementary DNA, the complementary DNA in aqueous solution is recognized, and the signal converter converts the DNA hybridization signal into a signal that can be analyzed.

In order to efficiently detect DNA hybridization with a DNA chip, an effective surface treatment is required to increase the hybridization efficiency and to remove the background caused by nonspecific binding. Much research has been conducted to make such surface-treated DNA chip platforms (Anal. Biochem., 266: 23-30, 1999; Nuc. Acid. Res., 29 (21): 107, 2001).

On the other hand, the application of CNTs in the field of biotechnology has recently appeared. Glucose chip, detection of proteins, detection of specific DNA sequences (Anal. Bioanal. Chem., 375: 103-5, 2003; Proc. Natl. Acad. Sci. USA, 100 (9): 4984-9, 2003; Anal. Bioanal. Chem., 375: 287-93, 2003). Biomolecular screening in CNT-based multilayers can increase the amount of biomolecules such as DNA immobilized due to its large surface area and excellent electrical conductivity, and increase detection sensitivity to biomolecules.

Currently, the method of detecting reaction results in biochips is the most common method using conventional fluorescent materials and isotopes (Toriba, A. et al. , Biomed. Chromatography: BMC., 17: 126-32, 2003; Raj, SU et al. , Anal. Chim. Acta, 484: 1-14, 2003; Peggy, AT et al. , J. Microbio.Meth., 53: 221-33, 2003), more easily and accurately As new methods of measurement are attempted, the need for a new material called CNT is growing.

PNA (Petide Nucleic Acid: DNA Analogue) A site-specific fixation of single-walled CNTs and a report that detected complementary binding to the target DNA (Williams, KA et al. , Nature, 420: 761) , 2001) and electrochemical methods to fix oligonucleotides to CNT arrays, and to detect DNA through guanidine oxidation (Li, J. et al. , Nano Lett., 3: 597-). 602, 2003). However, these did not apply CNT to the manufacture and development of biochips.

Recently, a high capacity biomolecular detection sensor (WO 03/016901 A1) using CNTs is known. The present invention relates to a multi-channel biochip obtained by arranging a plurality of CNTs by attaching a plurality of CNTs on a substrate and attaching various kinds of receptors.

On the other hand, the conventional DNA chip in which the oligonucleotide or the DNA is immobilized on the amine-treated substrate has a disadvantage in that it is difficult to uniformly attach DNA to the surface due to the uneven surface distribution of the amine group treated on the substrate. In addition, when analyzing the results of the hybridization reaction using a fluorescent material, there is a disadvantage that a lot of unnecessary signals are generated.

Accordingly, the present inventors fixed the CNTs with high density through chemical bonding on the substrate exposed to the amine group to form a high density CNT film exposed to the carboxyl group, and then prepared a CNT-DNA chip in which DNA was bound to the CNT film. As a result of applying to the hybridization reaction, it was confirmed that the unnecessary signal was significantly reduced when fluorescence observation and the fluorescence intensity to be detected were very strong and the present invention was completed.

SUMMARY OF THE INVENTION An object of the present invention is to provide a CNT film having high density fixed by chemical bonding on a substrate and having a chemical functional group exposed on the surface thereof, and a CNT-biochip having a bioreceptor attached to the CNT film surface.

Another object of the present invention is to provide a method for detecting various target biomaterials that bind or react with various types of bioreceptors using the CNT-biochip.

Still another object of the present invention is to provide a CNT-DNA chip having DNA attached to a CNT film fixed at high density by chemical bonding on a substrate, and a method for detecting a DNA hybridization reaction, wherein the DNA chip is used. ,

In order to achieve the above object, the present invention comprises the steps of (a) reacting the substrate exposed to the amine group and the CNT exposed carboxyl group to form a CNT monolayer on the surface of the substrate through the amide bond between the amine group and the carboxyl group; (b) reacting the CNT monolayer with a diamine-based organic compound to form an organic amine layer on the CNT monolayer, and stacking the CNTs by reacting the organic amine with the carboxyl group-exposed CNT; And (c) repeating step (b) n times to stack the CNT layer and the organic amine layer n times to form a high-density CNT film exposed to the carboxyl group, wherein the carboxyl group is exposed to the surface. It provides a method for producing a high density CNT film.

The present invention also provides a high-density CNT film produced by the above method and having a carboxyl group exposed on its surface.

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The present invention also comprises the steps of (a) reacting a substrate having an amine group exposed on the surface with a CNT exposed carboxyl group to form a CNT monolayer on the surface of the substrate through an amide bond between the amine group and the carboxyl group; (b) reacting the CNT monolayer with a diamine-based organic compound to form an organic amine layer on the CNT monolayer, and stacking CNTs by reacting the organic amine with a CNT exposed carboxyl group; (c) repeating step (b) n times to alternately stack the CNT layer and the organic amine layer n times to form a dense CNT film exposed to the carboxyl group; And (d) modifying the dense CNT film having the carboxyl group exposed to a chemical substance having a functional group which combines the carboxyl group with a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group, and a halogen. It provides a method for producing a high-density CNT film exposed to the chemical functional group selected from the group consisting of an amine group, aldehyde group, hydroxyl group, thiol group and halogen on the surface characterized in that it comprises.

In the present invention, a chemical having a functional group which combines with a carboxyl group and a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group and a halogen is selected from H ₂ NR ₁ -NH ₂ , H ₂ NR ₂ -SH, It may be characterized as H ₂ NR ₃ -OH or H ₂ NR ₄ -CHO. Where R ₁ , R ₂ , R ₃ and R ₄ are independently C _1-20 saturated hydrocarbons, unsaturated hydrocarbons or aromatic organic groups.

The present invention also provides a high-density CNT film prepared by the above method and having a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group and a halogen exposed on a surface thereof.

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In the present invention, the substrate may be characterized in that the material selected from the group consisting of silicon, glass, molten silica, plastic and PDMS, the substrate exposed to the amine functional group on the surface is obtained by treating the substrate with aminealkyloxysilane Can be.

In this invention, it is preferable that the chemical functional group couple | bonded with a carboxyl group is an amine group or a hydroxyl group.

In the present invention, the bioreceptor is an enzyme substrate having a carboxyl group, an amine group, a hydroxyl group, an aldehyde group, or a thiol group, a ligand, an amino acid, a peptide, a protein, a nucleic acid (DNA, RNA, PNA), a lipid, a cofactor or a carbohydrate. You can do

In the present invention, the target biomaterial is a substance capable of serving as a target to be detected by reacting with or binding to a bioreceptor, preferably a protein, nucleic acid, antibody, enzyme, carbohydrate, lipid or other bio-derived biomolecule, More preferably nucleic acid or protein.

The term 'CNT-biochip' used in the present invention encompasses a concept in which a bioreceptor is chemically or physicochemically bonded to a CNT film. The bioreceptor is attached to a high-density CNT film by chemical or physicochemical bonding. It is defined as including a sensor.

In the present invention, CNTs are repeatedly laminated on a solid substrate coated with a chemical functional group (amine group) through chemical bonding to prepare a CNT film exposed to a carboxyl group having a high surface density, and then reacted chemically or physicochemically with the carboxyl group. Bioreceptors having functional groups (amine groups, hydroxyl groups, and the like) are attached to fabricate CNT-biochips that can directly detect various types of target biomaterials or detect them using electrochemical signals.

In addition, by forming a high-density CNT film exposed to the amine group by the modification of the carboxyl group exposed on the surface of the CNT film, the bioreceptor having a carboxyl group or an aldehyde group reacting with the amine group is chemically or physicochemically bonded to produce a biochip.

On the other hand, in order to attach a bioreceptor having no functional group that binds the carboxyl group or an amine group, a chemical functional group that binds the CNT film exposed to the carboxyl group and a functional group that binds to the functional group possessed by the target bioreceptor at the same time Eggplants can be modified using chemicals. Thus, almost all bioreceptors can be chemically or physicochemically bound to the high density CNT film.

For example, in order to attach a bioreceptor having a thiol group, the thiol functional group is exposed to the surface of the CNT film through modification of the CNT surface using a chemical functional group that binds a carboxyl group and a chemical compound having a thiol functional group, and then the SS bond is used to attach the bioreceptor. A bioreceptor having a thiol group can be attached.

In addition, according to the present invention, there are abundant chemical functional groups on the surface of the high density CNT film, and thus it is possible to attach the bioreceptor at high density, and the unnecessary fluorescence signal is reduced during the fluorescence measurement of the hybridization reaction using the CNT-DNA chip. Can be detected.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

In particular, the following examples illustrate only DNA as a bioreceptor, but enzyme substrates, ligands, amino acids, peptides, proteins, RNA, PNA, lipids, noses having chemical functional groups such as carboxyl groups, amine groups, hydroxyl groups, aldehyde groups and thiol groups It will be apparent to those skilled in the art to fabricate CNT-biochips by fixing bioreceptors such as factor, carbohydrate and the like to the CNT film.

Example 1 Preparation of CNT Exposed to Carboxyl Group

The CNTs used in the present invention are not particularly limited, and commercially available products may be purchased and used, or may be manufactured and used by conventional methods. Pure CNTs must be carboxylated on the surface and / or both ends of the CNTs for use in the present invention.

The CNT exposed carboxyl group was refluxed for 24 hours in a sonicator containing a strong acid (mixture of nitric acid and sulfuric acid) solution, filtered through a 0.2 μm filter, and then the filtrate was immersed in jinsan. It was refluxed for hours and then centrifuged. The supernatant was recovered, filtered through a 0.1 μm filter and dried. CNTs exposed to the dried carboxyl group were dispersed in distilled water or an organic solvent, and then filtered through a 0.1 μm filter to select CNTs having a constant size.

Example 2: Preparation of Substrate Exposed Amine Group

In the present invention, the amine alkyloxysilane is fixed on the surface of a substrate such as silicon, glass, molten silica, plastic, PDMS (polydimethylsiloxane), and the amine group is exposed on the surface of the substrate, but a substrate surface-treated with a commercially available amine is purchased. It is also possible to use.

Example 3 Preparation of High Density CNT Film Exposed to Surface of Carboxyl Group

The CNT exposed carboxyl group prepared in Example 1 was reacted with the exposed amine group prepared in Example 2 to form a CNT monolayer on the substrate through an amide bond between the carboxyl group and the amine group (FIG. 1A).

Next, the CNT attached to the substrate with an amide bond reacts with the diamine organic compound having a double amine functional group, and the CNT exposed carboxyl group is reacted with the other amine group of the diamine organic compound to form an amide bond. CNTs were laminated (FIG. 1B).

Subsequently, the chemical reaction between the CNT exposed to the carboxyl group and the diamine organic compound was repeatedly performed to prepare a high density CNT film in which the CNT layer and the organic amine layer were alternately stacked n times and the carboxyl group was exposed to the surface ( 1c).

In the present invention, a substance having a chemical formula of H ₂ NR ₁ -NH ₂ may be used as the diamine organic compound. Here, R ₁ is C _1-20 saturated hydrocarbons, unsaturated hydrocarbons or aromatic organic groups.

In order to promote the amide bond, HAMDU ( O- (7-azabenzotriazol-1-yl) -1,3-dimethyl-1,3-dimethyleneuronium hexafluorphosphate), DCC (1,3-dicyclohexylcarbodiimide), HAPyU ( O- (7-azabenzotriazol-1-yl) -1,1: 3,3-bis (tetramethylene) uronium hexafluorphosphate), HATU ( O- (7-azabenzotriazol-1-yl) -1,1: 3,3- tetramethyluronium hexafluorphosphate), HBMDU ( O- (benzotriazol-1-yl) -1,3-dimethyl-1,3-dimethyleneuronium hexafluorphosphate), HBTU ( O- (benzotriazol-1-yl) -1,1,3,3- It is preferable to use tetramethyluronium hexafluoro phosphate (DI) (diisopropylethylamine), TMP (2,4,6-trimethylpyridine), NMI ( N- methylimidazole) or the like as a base.

When water is used as a solvent, EDC (1-ethyl-3- (3-dimethylamini-propyl) arbodiimide hydrochloride) is used as a coupling agent, and NHS ( N -hydroxysuccinimide), NHSS (N- hydroxysulfosuccinimide) is preferable.

In this example, HATU was used as the coupling agent and DIEA was used as an auxiliary agent of the coupling agent.

The coupling agent serves to form an amide bond (-CONH-) with a -COOH functional group and a -NH ₂ functional group, and the base and the auxiliary agent help to increase efficiency when the coupling agent forms an amide bond. do.

Example 4 Preparation of High Density CNT Film Exposed with Amine Group on Surface

Modification of the surface of the CNT exposed carboxyl group is a chemical functional group (amine group, hydroxyl group, etc.) that reacts with the carboxyl group and chemical functional groups (amine group, hydroxyl group, thiol group, aldehyde group, etc.) that combine with the functional group possessed by the target bioreceptor It can be reformed using a chemical having both at the same time. Chemicals that can be used for the modification include H ₂ NR ₁ -NH ₂ , H ₂ NR ₂ -SH, H ₂ NR ₃ -OH, H ₂ NR ₄ -CHO Etc. can be used. Where R ₁ , R ₂ , R ₃ and R ₄ are independently C _1-20 saturated hydrocarbons, unsaturated hydrocarbons or aromatic organic groups.

In this embodiment, the carboxyl group of the CNT film having a carboxyl group exposed on the surface prepared in Example 3 and the amine group of the chemical represented by H ₂ NR ₁ -NH ₂ are bonded to prepare a CNT film finally exposed to the surface of the amine group. It was.

Example 5 Preparation of DNA Chips Fixing DNA on CNT Film Exposed to Carboxyl Group

A DNA chip was prepared by chemically binding DNA to a CNT film having a carboxyl group exposed on its surface. A CNT-DNA chip was prepared by binding an amino group of DNA to a CNT film having a carboxyl group exposed in Example 3 (FIG. 2). In this embodiment, EDC was used as a coupling agent for the amide bond of the carboxyl group and the amino group, and NHS was used as an auxiliary agent of the coupling agent. In this example, a CNT-DNA chip was prepared using the oligonucleotide of SEQ ID NO: 1 having an amino group at its terminal.

SEQ ID NO: 5'-TGT GCC ACC TAC AAG CTG TG-3 '

The presence of DNA on the CNT film was confirmed by X-ray photoelectron spectroscope (XPS) spectra for phosphorus atoms using the fact that all DNA contained phosphate groups (FIG. 4). . As shown in FIG. 3, phosphorus was detected in the XPS surface analysis, and it was confirmed that DNA was bound to the CNT surface.

Example 6: Preparation of DNA Chips Fixing DNA on CNT Film Exposed to Amine Group

In this example, a CNT-DNA chip was prepared by binding a carboxyl group of DNA to a CNT film exposed to an amine group on the surface prepared in Example 4 (FIG. 3). In this embodiment, EDC was used as a coupling agent for the amide bond of the carboxyl group and the amino group, and NHS was used as an auxiliary agent of the coupling agent. In this embodiment, a CNT-DNA chip was prepared using the oligonucleotide of SEQ ID NO: 1 having a carboxyl group at the terminal.

Example 7: Analysis of hybridization reaction using DNA chip immobilized DNA on CNT film exposed carboxyl group

The CNT-DNA chip prepared in Example 5 was placed in a hybridization chamber, and then the hybridization solution was pipetted down and covered with a cover slide. At this time, the hybridization solution is filled with a solution containing 32 μl of complementary nucleotide oligonucleotides, and the final concentration is 3XSSC (0.45M NaCl, 0.045M sodium citrate), 0.3% sodium dodecyl sulfate (SDS), and the total volume. To 40 μl. Here, the complementary oligonucleotide sequence of the hybridization solution was used in the following SEQ ID NO: 2 having a fluorescein isothiocyanate (FITC) attached to the terminal.

SEQ ID NO: 5'-CAC AGC TTG TAG GTG GCA CA-3 '

To remove nonspecific binding of the two oligonucleotide strands, the solution was left at 100 ° C. for 2 minutes and then centrifuged at 12000 rpm for 2 minutes. In order to prevent the hybridization solution from drying in the hybridization chamber, 30 μl of 3 × SSC (0.45 M NaCl, 0.045 M sodium citrate) was added to the depressions on both sides of the chamber. The chamber lid was closed and left in a 55 ° C. thermostat for 10 hours.

After 10 hours, the hybridization chamber was removed from the thermostat and soaked in 2XSSC solution for 2 minutes, then for 5 minutes in 0.1XSSC (0.015M NaCl, 0.0015M sodium citrate) and 0.1% SDS solution, and finally 5 Soak for minutes. In order to remove the remaining solution on the DNA chip, the chip was placed in a centrifuge and centrifuged at 600 rpm for 5 minutes.

Hybridization reactions detected fluorescence using FITC labeled at the oligonucleotide terminus of SEQ ID NO: 2. Fluorescence images were obtained using ScanArray 5000 Packard BioScience, BioChip Tecnologies LLC confocal microscopy and QuantArray Microarray Analysis Software (FIG. 5).

FIG. 5 (a) shows a fluorescence image of a high density CNT film before binding DNA, and in FIG. 5 (b), (1) hybridizes complementary DNA after binding DNA having amino end groups on the CNT film. The fluorescence detection result in the case of reaction is shown, and (2) shows the fluorescence detection result in the case of hybridizing non-complementary DNA. When hybridizing the oligonucleotide having a nucleotide sequence complementary to the oligonucleotide bound to the CNT film, it was confirmed that the fluorescence appeared clearly and evenly ((1) of FIG. 5 (b)). However, when the oligonucleotide is not fixed CNT film (Fig. 5 (a)) and the oligonucleotide having a base sequence not complementary to the oligonucleotide bound to the CNT film hybridized (Fig. 5 (b) (2)) showed no fluorescence at all. From this result, it was confirmed that almost no specific reaction occurred.

Example 8 Analysis of Hybridization Reaction Using DNA Chips Fixing DNA on CNT Film Modified with Amine Group

The same hybridization reaction as in Example 7 was performed using the CNT-DNA chip prepared in Example 6 (FIG. 6). In FIG. 6, (a) shows fluorescence detection results when binding complementary DNA, and (b) shows fluorescence detection results when binding non-complementary DNA. As shown in FIG. 6, oligonucleotides having non-complementary nucleotide sequences and oligonucleotides having complementary nucleotide sequences were clearly distinguished from hybridized ones and hybridized ones.

As described in detail above, the present invention is obtained by repeatedly fixing the CNT exposed carboxyl group on the substrate exposed amine group through an amide bond and a high-density CNT film exposed to the chemical functional group on the surface and the bio There is an effect of providing a biochip in which the receptor is chemically or physicochemically bonded.

According to the present invention, it is possible to manufacture a CNT-biochip in which various bioreceptors are chemically or physicochemically bonded to a CNT film having a carboxyl group exposed thereto or a CNT film having the carboxyl group modified with various chemical functional groups. In addition, it is possible to produce a CNT-biochip in which the bioreceptor is uniformly and densely adhered to the surface of the CNT film because the CNT film is rich in chemical functional groups and is evenly present.

In particular, when the fluorescence measurement of DNA hybridization reaction using the CNT-DNA chip according to the present invention can reduce the unnecessary signal very excellent results can be obtained. Such CNT-DNA chips are useful for genotyping, mutation detection, pathogen identification, and the like.

<110> KAIST <120> Carbon Nanotube Film with High Surface Density and A Biochip          Using the same <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> oligonucleotide for DNA-chip <400> 1 tgtgccacct acaagctgtg 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> oligonucleotide for DNA-chip <400> 2 cacagcttgt aggtggcaca 20

Claims (19)

Method for producing a high density CNT film having a carboxyl group exposed on the surface comprising the following steps: (a) reacting a substrate having an amine group exposed to a surface with a CNT having a carboxyl group exposed thereto to form a CNT monolayer on the surface of the substrate through an amide bond between the amine group and the carboxyl group; (b) reacting the CNT monolayer with a diamine-based organic compound to form an organic amine layer on the CNT monolayer, and stacking CNTs by reacting the organic amine with CNT exposed carboxyl groups; And (c) repeating step (b) n times to alternately stack the CNT layer and the organic amine layer n times to form a high density CNT film exposed to the carboxyl group. Method for producing a high-density CNT film exposed to a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group and a halogen on the surface comprising the following steps: (a) reacting a substrate having an amine group exposed to a surface with a CNT having a carboxyl group exposed thereto to form a CNT monolayer on the surface of the substrate through an amide bond between the amine group and the carboxyl group; (b) reacting the CNT monolayer with a diamine-based organic compound to form an organic amine layer on the CNT monolayer, and stacking CNTs by reacting the organic amine with CNT exposed carboxyl groups; (c) repeating step (b) n times to alternately stack the CNT layer and the organic amine layer n times to form a dense CNT film exposed to the carboxyl group; And (d) reforming the dense CNT film exposed to the carboxyl group with a chemical having a functional group which binds the carboxyl group and a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group and a halogen. The method of claim 1 or 2, wherein the substrate is a material selected from the group consisting of silicon, glass, fused silica, plastic, and PDMS. The method of claim 1 or 2, wherein the substrate having an amine functional group exposed to the surface is obtained by treating the substrate with aminealkyloxysilane. The method according to claim 1 or 2, wherein the chemical functional group which binds to the carboxyl group is an amine group or a hydroxyl group. The chemical substance according to claim 2, wherein the chemical substance which has a functional group which binds a carboxyl group and a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group, and a halogen at the same time is H ₂ NR ₁ -NH ₂ , H ₂ NR ₂ -SH , H ₂ NR ₃ -OH or H ₂ NR ₄ -CHO, wherein R ₁ , R ₂ , R ₃ and R ₄ are independently C _1-20 saturated hydrocarbons, unsaturated hydrocarbons or aromatic organic groups). A high density CNT film produced by the method of claim 1 and having a carboxyl group exposed on its surface. A high density CNT film prepared by the method of claim 2, wherein a chemical functional group selected from the group consisting of an amine group, an aldehyde group, a hydroxyl group, a thiol group, and a halogen is exposed on a surface thereof. delete delete delete delete delete delete delete delete delete delete delete

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