KR20050033312A - Method for fabricating a nano-biochip using the nanopattern of block copolymers - Google Patents

Abstract

A method for fabricating a nano-biochip by using nano-pattern of block copolymers is provided, thereby immobilizing real-life samples on the surface of the nanometer-sized biochip with high density without loss of tertiary structure of a protein in the real-life samples. The method for fabricating a nano-biochip by using nano-pattern of block copolymers comprises the steps of: (a) forming a metal thin layer with affinity to a bio-receptor on a substrate; (b) coating the metal thin layer with the block copolymers; (c) heating the block copolymers to induce self-assembly of the copolymers and form a regular structure; (d) etching the copolymers to form a porous nano-pattern; and (e) attaching the bio-receptor to the metal exposed by the porous nano-pattern, wherein the bio-receptor is an enzyme substrate, a ligand, an amino acid, a peptide, a protein, a nucleic acid, a lipid, a cofactor or a carbohydrate; the metal with affinity to the bio-receptor is Au, Ag, Pt, Nb, Ta, Zr or alloy of Co and Cr; and the block copolymer is polystyrene-polymethylmethacrylate(PS-PMMA), polystyrene-polybutadiene(PS-PB) or polystyrene-polyisoprene(PS-PI).

Description

Method for fabricating a nano-biochip using the nanopattern of block copolymers

The present invention manufactures a regular nanopattern using a self-assembly of the block copolymer (block copolymer), and using a nanopattern of the polymer to prepare a nanopattern of a metal having affinity with a bioreceptor Then, the present invention relates to a method for manufacturing a nano-biochip, characterized in that selectively attaching a bioreceptor that binds a target biomaterial to the metal of the nanopattern. The present invention also relates to a nano-biochip manufactured by the above method and a method for detecting various target biomaterials that bind to or react with a bioreceptor using the same.

Recently, in order to manufacture patterns of 100 nm or less, various attempts have been made to manufacture new fine patterns instead of the conventional photoetching process. In semiconductor processes, miniaturization and high integration of devices is an important process for reducing time, cost, sample size, and improving new functions.

However, it is known that it is difficult to manufacture a pattern of 100 nm or less due to the limitation of resolution in the photolithography process which is the current semiconductor process. In addition, until recently, many memory products have increased the magnetic density by increasing the magnetic anisotropy of the magnetic material. This method becomes impossible due to the thermal instability (superparamagnetic limit) problem in the continuous magnetic thin film.

Accordingly, in order to manufacture fine patterns of 100 nm or less, an electron beam (E-beam), an X-ray (x-ray) etching method, and the like have been studied, but have long problems such as expensive equipment and processing time. When the self-assembly of the block copolymer is used, it is inexpensive and easy to process, and the size and structure of the pattern can be controlled by the nature and size of the block copolymer.

Recently, research has been actively conducted to prepare micropatterns of several nanometers to several tens of nanometers using self-assembly of block copolymers (Science, 276: 1401-4; Nature, 405: 433-7; Adv). Mater., 12: 787-91). However, the research on metal nanopatterns using the same has a problem that there is a limit in aspect ratio of the pattern or the type of materials used, and in particular, a study on nano-biochips in which bioreceptors are coupled to metal nanopatterns Was not.

On the other hand, 10,000 human genes are currently predicted, of which about 10,000 functions have been revealed, 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, developed a method for treating and preventing diseases that were not possible with classical methods. Essential is an efficient protein-protein and protein-ligand reaction detection technique.

The protein-protein reaction detection technique using a protein chip regulates the orientation of biomolecules at the molecular level by using an affinity tag for a target protein, thereby depositing a single layer of uniform and stable protein on the surface of the support. After specific fixation, the protein-protein interactions are analyzed (Paul, J. et al., JACS, 122: 7849-50, 2000; RaVi, A. et al., Anal. Chem., 73: 471 -80, 2001; Benjamin, T. et al., Tibtech., 20: 279-81, 2002).

Numerous genetic information gained by the Human Genome Project has paved the way for innovation in understanding and diagnosing genetic diseases. In this context, development of effective DNA identification systems for genomic sequencing, mutation detection, and pathogen identification is required.

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. 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.

As signal changers, optical, piezoelectric and electrochemical reactions have been studied. Among them, the electrochemical reaction is characterized by high sensitivity, low cost, and compatibility with microfabrication technology, and can detect DNA having a specific sequence quickly and directly. Has the characteristics.

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).

In addition, various methods for detecting DNA hybridization have been sought, such as the scanmetric method, the calorimetric method, the method using nanoparticles, and the method using electrochemistry (Science, 289: 1757). -60, 2000; Anal. Biochem., 295: 1-8, 2001; Analyst., 127: 803-8, 2002; Anal. Bioanal. Chem., 375: 287-93, 2003).

On the other hand, microarray protein chips (microarray) is currently occupying a large proportion of research on diagnostic protomics (diagnostic protomics). When arraying polypeptides on the surface of a substrate, early array techniques (USP 5,143,854A), which used photolithographics, have recently been tried in various ways. In particular, the development of microarray-type formats is becoming increasingly important in a variety of immunoassays, including antigen-antibody pairs and enzyme-liked immunosorbent assays. have. In addition, a patent has recently been filed for comparing relative content of biomolecules and identifying biomolecules in a sample using affinity tags and mass spectrometry (WO 2002/86168 Al).

However, protein arrays are difficult to integrate or array in a practical format that is smaller or more sensitive than DNA arrays. That is, the lattice pattern of DNA oligonucleotides can be generated on the surface of the substrate by photoetching techniques, but for proteins consisting of hundreds of amino acids, the antibody should generally have about 1400 amino acids for accurate diagnosis of the disease. In order to achieve a higher density of the grid pattern is required, but this is not easy to realize.

Another problem is that the protein's tertiary structure can easily be lost when dealing with proteins under denaturing conditions ( Anal. Chem . 14A-15A, 2001; Anal. Chem., 73 , 8-12, 2001). There are many limitations when operating.

The solution to these problems depends on how high the protein is arranged without losing the tertiary structure of the protein. To date, inkjet printing, drop-on-demand Technology, microcontact printing, and IBM's soft lithography. However, these methods also have spacing sizes of tens of micrometers to several millimeters, and dozens of nanometer-sized protein chips with high density of real-life samples attached without losing the tertiary structure of the protein. Development has never been attempted.

Accordingly, the present inventors have made diligent efforts to develop nano-biochips using an ultra-high density pattern having a simpler process and a pattern size of several tens of nanometers. As a result, regular nanopatterns using self-assembly of block copolymers are used. The present invention was completed by forming a nanopattern of a metal having affinity with a bioreceptor, and then fixing a bioreceptor that binds to or reacts with a target biomaterial.

Disclosure of Invention An object of the present invention is to provide a nano-biochip having various kinds of bioreceptors attached to a nanopattern of a metal formed using a block copolymer nanopattern, and a method of manufacturing the same.

Still 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 nano-biochip.

In order to achieve the above object, the present invention comprises the steps of (a) forming a thin film of metal having affinity with the bioreceptor on the substrate; (b) applying a block copolymer onto the thin film; (c) heat treating the block copolymer to induce self-assembly and form a regular structure; (d) etching the block copolymer in which the regular structure is formed to form a porous nanopattern; And (e) attaching a bioreceptor to the metal exposed by the porous nanopattern, wherein the bioreceptor is attached to the metal exposed by the nanopattern of the block copolymer. Provide a method.

The present invention also provides a nano-biochip and the nano-biochip, which are manufactured by the above method and have a bioreceptor attached or reacted with a target biomaterial to a metal exposed by the nanopattern of the block copolymer. It provides a method for detecting a target biomaterial that binds to or reacts with a bioreceptor, characterized in that using a.

The present invention also comprises the steps of (a) forming a thin film of metal (M) having affinity with the bioreceptor (BR) on the substrate (S); (b) applying a block copolymer onto the thin film; (c) heat treating the block copolymer to induce self-assembly and form a regular structure; (d) etching the block copolymer in which the regular structure is formed to form a porous nanopattern; (e) selectively depositing a mask metal on the porous nanopattern; (f) etching the thin film of a metal having affinity with the block copolymer and the bioreceptor BR using the deposited metal as a mask to form a nanopattern of metal on a substrate; And (g) attaching the bioreceptor to a metal having an affinity with the bioreceptor, the method for producing a nano-biochip of S-M-BR type.

The present invention also provides a SM-BR type, characterized in that the bioreceptor (BR) is prepared by the above method, the metal (M) nanopattern on the substrate (S) is attached to or react with the target biomaterial. The present invention provides a method for detecting a target biomaterial that binds to or reacts with a bioreceptor using a nano-biochip and the nano-biochip.

The heat treatment of the block copolymer to induce self-assembly and to form a regular structure may be characterized by slow cooling after raising the temperature above the liquid crystal phase transition temperature of the used block copolymer.

In the present invention, the block copolymer may be polystyrene-polymethylmethacrylate (PS-PMMA), polystyrene-polybutadiene (PS-PB), or polystyrene-polyisoprene (PS-PI). Copolymers that cause self-assembly can be used without limitation.

The etching of the block copolymer having the regular structure to form the nanopattern of the block copolymer may be characterized in that the UV etching and then performing a reactive ion etching, the block metal on which the mask metal is deposited Etching the thin film of the block copolymer and the metal using the regular structure of the coal as a mask to form a nanopattern of the metal on the substrate may be characterized by using reactive ion etching and ion milling.

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.

In the present invention, the bioreceptor is a substance capable of reacting with or binding to the target biomaterial, and may be an enzyme substrate, a ligand, an amino acid, a peptide, a protein, a nucleic acid, a lipid, a cofactor, or a carbohydrate. It may be characterized as a protein or DNA related to the disease. In addition, they may be characterized by having a thiol group.

In the present invention, the metal having affinity with the bioreceptor is gold (Au), silver (Ag), platinum (Pt), niobium (Nb), tantalum (Ta), zirconium (Zr), or cobalt (Co) and chromium It is characterized by being a alloy of (Cr), Preferably it is characterized by being gold. It is also preferable to use the same mask metal as the metal having affinity with the bioreceptor formed on the substrate.

Gold can selectively bind to bioreceptors (peptides, proteins, DNA, etc.) containing thiol groups (-SH). By using these properties, bioreceptors can be selectively coupled to gold nanopatterns to fabricate high-density nano-biochips.

The term 'nano-biochip' used in the present invention encompasses a concept in which a bioreceptor that binds to or reacts with a biomaterial is attached to a metal nanopattern, and is defined as including a nano-biosensor and a nano-bioarray. .

As a method of detecting a target biomaterial that binds to a bioreceptor using the nano-biochip of the present invention, a method using a conventionally known electrical detection method and a resonance method may be used.

Specifically, by attaching a radioisotope to the reactants, the radiation may be measured using a measuring instrument at a certain surface after the reaction, and the amount of redox reaction and charge accumulation may be measured in a liquid phase using electrical properties. It is also possible to use a method using. Redox reactions are currently common electrochemical detection methods that can measure changes in hydrogen or electrons using cyclic voltametry, potentiometry, amperometry, and the like.

Block copolymers are macromolecules having a larger or more complex structure in which two or more chemically different blocks are bonded to each other by covalent bonds, and have a phase-separated structure depending on the chemistry of the block, the length (or molecular weight) of the block, and the volume composition. And appear differently and generally have a pattern size between about ten and one hundred nanometers. For example, diblock copolymers have lamellar, gyroid, cylindrical, spherical, etc., depending on the composition ratio. Many different structures can be formed.

Properties of copolymers suitable for the purposes of the present invention are as follows.

Firstly, the phase separation should occur spontaneously with a chemically different structure, and it must have a spherical structure in order to make metal points in the shape of a plate, cylinder or sphere. Plate-shaped and cylindrical structures are available for the purpose of producing metal wires.

Second, the etching selectivity between the different blocks must be different so that one side can be easily removed selectively. Examples of such copolymers include polymers in which one block containing polystyrene-polymethylmethyl acrylate (PS-PMMA) is an acrylate and polymers in which one block containing polystyrene and polybutadiene (PS-PB) have a double bond in the main chain. Is applicable. In addition, when one block is an inorganic polymer including a metal or ceramic and the other is an organic polymer, one side can be easily removed by using a difference in resistance to heat or plasma.

One block of the copolymer used in the present invention, polymethyl methacrylate (PMMA), polybutadiene (PB), or polyisoprene (PI), is selectively removed by UV, providing a desired level of thin film thickness and a suitable pattern. Reactive ion etching (RIE) using plasma is used.

One of the characteristics of the present invention is that the desired metal is deposited in advance to form a thin film and then selectively etched through a patterning process, compared to conventional methods for selectively growing metal after forming a pattern. It has the following advantages.

First, patterning is possible after adjusting the properties of the desired metal in advance.

Secondly, in the case of selectively depositing a desired metal after forming an existing pattern, it is difficult to produce a metal point having a high aspect ratio due to the limitation of polymer pattern depth and the limitation of selective deposition. By forming and depositing the mask metal, there is an advantage that the desired thickness of the thin film can be freely adjusted and a metal pattern having a higher aspect ratio can be made according to the nature of the mask metal.

According to the present invention, a pattern of less than 100 nanometers, which is difficult to implement in a commercially available photolithography process, is possible, and complicated process equipment is not required, and because the process is relatively simple, the process cost is low. In addition, the size of the polymer pattern can be adjusted according to the size and the degree of etching of the polymer block used, and thus the size of the metal dots can be adjusted from a few to several tens of nanometers.

In order for the present invention to be effective, it is preferable to apply the following special treatment.

First, in order to increase the adhesion between the polymer and the metal, the surface of the target metal is treated with oxygen plasma or the like before coating the block copolymer.

Second, in order to effectively remove the polymer, ion milling is first performed and oxygen plasma etching is performed. Because some metal is deposited on top of the polymer, direct oxygen plasma etching cannot remove the polymer.

Hereinafter, with reference to the accompanying drawings will be described the present invention in more detail.

[First Embodiment]

1 is a schematic diagram of a method of manufacturing a nano-biochip by attaching a bioreceptor to a metal exposed by a nanopattern of a block copolymer according to a first embodiment of the present invention.

First, a thin film of a metal having affinity with a bioreceptor is formed on a substrate, and a block copolymer capable of self-assembly is coated thereon (FIG. 1 (a)).

Next, the self-assembly of the block copolymer is induced by heat treatment to form a regular structure (Fig. 1 (b)).

Next, UV etching is performed to form a porous nanopattern (FIG. 1 (c)), and a nanopattern exposed to a metal having affinity with a bioreceptor is exposed through reactive ion etching (FIG. 1 (d)). ).

Finally, a bio-receptor is coupled to the metal to prepare a nano-biochip (FIG. 1 (e)).

Second Embodiment

2 is a schematic diagram of a method of manufacturing a nano-biochip by attaching a bioreceptor to a metal nanopattern on a substrate according to a second embodiment of the present invention.

First, a thin film of metal having affinity with the bioreceptor is formed, and a block copolymer capable of self-assembly is coated thereon (FIG. 2 (a)).

Next, the self-assembly of the block copolymer is induced by heat treatment to form a regular porous structure (FIG. 2 (b)).

Next, UV etching is performed to form a porous nanopattern (FIG. 2 (c)), and through a reactive ion etching to form a nanopattern to which a metal having affinity with the bioreceptor is exposed (FIG. 2 (d)). ).

Next, selectively depositing a metal serving as a mask on the nanopattern (FIG. 2 (e)) and performing reactive ion etching and ion milling to form a dot-shaped metal nanopattern on the substrate (FIG. 2 (f) )).

Finally, a bio-receptor is coupled to the metal to prepare a nano-biochip (Fig. 2 (g)).

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 the following examples, polystyrene-polymethylmethacrylate (PS-PMMA) was used as the block copolymer, but air inducing self-assembly such as polystyrene-polybutadiene (PS-PB) and polystyrene-polyisoprene (PS-PI) Coalescing may be used without limitation.

In addition, in the following examples, gold (Au) was used as a metal having affinity with a bioreceptor, but silver (Ag), platinum (Pt), niobium (Nb), tantalum (Ta), zirconium (Zr), or cobalt ( Metals with high polarization resistance, such as alloys of Co) and chromium (Cr), can be used without limitation.

In addition, although the DNA containing the thiol group is exemplified as the bioreceptor, the use of all bioreceptors such as peptides and proteins containing the thiol group will be included in the scope of the present invention.

In addition, in the following examples, only detection of hybridization reaction using a specific nano-DNA chip is illustrated, but from the knowledge in the field of biochips, a nano-biochip to which other bioreceptors are attached is manufactured to bind a bioreceptor and a target biomaterial. However, detecting the reaction will be apparent to those skilled in the art.

Example 1 Preparation of a Porous Polymer Pattern of 10-20 nm Levels

First, gold (Au) having affinity with a bioreceptor was deposited on a silicon wafer to a thickness of 20 nm. Next, in order to prepare a porous pattern on the order of ten to twenty nanometers, a spherical structure of polystyrene-polymethylmethacrylate (PS-PMMA) was spin coated at a speed of about 3000 rpm for 60 seconds to form a block copolymer polymer thin film. Formed (FIG. 1 (a)).

Then, heat treatment at about 160 ℃ to form a regular structure of the PS-PMMA (Fig. 1 (b)), and irradiated with a UV having a wavelength of 245nm polymethyl methacrylate (PMMA) removed porous After forming a nanopattern of (Fig. 1 (c)), a reactive ion etching was performed. In this process, gold is exposed to the portion from which the PMMA is removed to form a dot-shaped nanopattern (FIG. 1 (d)). 3 is a scanning electron micrograph of the nanopattern thus formed.

Example 2 Preparation of Metal Pattern of 10-20 nm Level

First, gold (Au) having affinity with a bioreceptor was deposited on a silicon wafer to a thickness of 20 nm. Next, in order to prepare a porous pattern on the order of ten to twenty nanometers, a spherical structure of polystyrene-polymethylmethacrylate (PS-PMMA) was spin coated at a speed of about 3000 rpm for 60 seconds to form a block copolymer polymer thin film. Formed (FIG. 2 (a)).

Next, heat treatment at about 160 ℃ to form a regular structure of the PS-PMMA (Fig. 2 (b)), and to irradiate UV having a wavelength of 245nm to form a porous nanopattern removed PMMA ( 2 (c)), reactive ion etching was performed. In this process, gold is exposed to a portion where the PMMA is removed to form a dot-shaped nanopattern (FIG. 2 (d)).

Next, gold (Au) is selectively deposited on the nanopattern of the polymer (FIG. 2 (e)), and then reactive ion etching using ion milling and oxygen plasma is performed to form a dot of gold (Au) on the substrate. ) Nanopattern was prepared (FIG. 2 (f)).

Example 3 Fabrication of Nano-Biochips

By binding the bioreceptor to the gold of FIG. 1 (d) obtained in Example 1 using the property that gold selectively binds to a bioreceptor (peptide, protein, DNA, etc.) containing a thiol group (-SH) Nano-biochips with bioreceptors attached to gold exposed by the nanopattern of the copolymer were prepared (FIG. 1 (e)). FIG. 1 (e) is a model of a nano-DNA chip in which DNA (oligonucleotide) is attached to gold exposed by the nanopattern of the block copolymer. The nano-DNA chip can be used for efficient detection of various hybridization reactions.

In the same manner, the S-Au-BR form in which the bioreceptor is coupled to the gold nanopattern formed on the substrate S by binding the bioreceptor BR to the gold (Au) of FIG. 2 (f) obtained in Example 2 Nano-biochips were prepared (FIG. 2 (g)). Figure 2 (g) shows a nano-DNA chip of the S-Au-DNA type DNA (oligonucleotide) attached to the gold nanopattern formed on the substrate. When using the nanoDNA chip, it is possible to efficiently detect a variety of hybridization reaction.

The present invention is a method for producing a new nano-biochip, characterized in that the bioreceptor coupled to a metal nanopattern of several tens of nanometers prepared using self-assembly of the block copolymer and a target bio to a metal pattern of tens of nanometers There is an effect of providing a nano-biochip that has a bioreceptor that binds to or reacts with a substance. The nano-biochip according to the present invention, in which the bioreceptor is integrated at a high density, may be usefully used to detect a reaction or binding between the bioreceptor and the target biomaterial.

1 is a schematic diagram of a method of manufacturing a nano-biochip by attaching a bioreceptor to a metal exposed by a nanopattern of a block copolymer according to a first embodiment of the present invention. (a) shows a coating of a block copolymer capable of self-assembly with a thin film of a metal having affinity with a bioreceptor on the substrate, and (b) shows a rule inducing self-assembly of the block copolymer by heat treatment. (C) shows the formation of a porous nanopattern by UV etching, and (d) shows the formation of a nanopattern with exposed metal having affinity with a bioreceptor through reactive ion etching. (E) represents a nano-biochip manufactured by coupling a bioreceptor to a metal.

2 is a schematic diagram of a method of manufacturing a nano-biochip by attaching a bioreceptor to a metal nanopattern on a substrate according to a second embodiment of the present invention. (a) shows a coating of a block copolymer capable of self-assembly with a thin film of a metal having affinity with a bioreceptor on the substrate, and (b) shows a rule inducing self-assembly of the block copolymer by heat treatment. (C) shows the formation of a porous nanopattern by UV etching, and (d) shows the formation of a nanopattern with exposed metal having affinity with a bioreceptor through reactive ion etching. (E) shows the selective deposition of a metal serving as a mask on the nanopattern, (f) shows the formation of a dot-shaped metal nanopattern on the substrate by performing reactive ion etching and ion milling. , (g) represents a nano-biochip manufactured by bonding a bioreceptor to the metal.

FIG. 3 is a scanning electron micrograph of the formation of a pattern using a block copolymer after forming a gold layer on a silicon wafer.

Claims (22)

A method of manufacturing a nano-biochip having a bioreceptor attached to a metal exposed by a nanopattern of the block copolymer, comprising the following steps: (a) forming a thin film of metal that is compatible with the bioreceptor on the substrate; (b) applying a block copolymer onto the thin film; (c) heat treating the block copolymer to induce self-assembly and form a regular structure; (d) etching the block copolymer in which the regular structure is formed to form a porous nanopattern; And (e) attaching the bioreceptor to the metal exposed by the porous nanopattern. According to claim 1, wherein the step (b) is characterized in that the slow cooling after raising the temperature above the liquid crystal phase transition temperature of the used block copolymer. The method of claim 1, wherein the bioreceptor is an enzyme substrate, ligand, amino acid, peptide, protein, nucleic acid, lipid, cofactor or carbohydrate. The method of claim 3, wherein the bioreceptor is a protein or DNA. The metal of claim 1, wherein the metal having affinity with the bioreceptor is gold (Au), silver (Ag), platinum (Pt), niobium (Nb), tantalum (Ta), zirconium (Zr), or cobalt (Co). And chromium (Cr) alloy. The method of claim 1, The block copolymer is polystyrene-polymethylmethacrylate (PS-PMMA), polystyrene-polybutadiene (PS-PB), or polystyrene-polyisoprene (PS-PI). The method of claim 1 wherein step (d) comprises performing a UV etch followed by a reactive ion etch. Method for manufacturing a nano-biochip of the S-M-BR type comprising the following steps: (a) forming a thin film of metal (M) having affinity with the bioreceptor (BR) on the substrate (S); (b) applying a block copolymer onto the thin film; (c) heat treating the block copolymer to induce self-assembly and form a regular structure; (d) etching the block block block in which the regular structure is formed to form a porous nanopattern; (e) selectively depositing a mask metal on the porous nanopattern; (f) etching the thin film of a metal having affinity with the block copolymer and the bioreceptor BR using the deposited metal as a mask to form a nanopattern of metal on a substrate; And (g) attaching the bioreceptor to the metal having affinity with the bioreceptor. The method of claim 8, wherein the step (c) is a slow cooling after raising the temperature above the liquid crystal phase transition temperature of the used block copolymer. The method of claim 8, wherein the bioreceptor is an enzyme substrate, a ligand, an amino acid, a peptide, a protein, a nucleic acid, a lipid, a cofactor or a carbohydrate. The method of claim 10, wherein the bioreceptor is a protein or DNA. The metal of claim 8, wherein the metal has affinity with the bioreceptor is gold (Au), silver (Ag), platinum (Pt), niobium (Nb), tantalum (Ta), zirconium (Zr), or cobalt (Co). And an alloy of chromium. 9. The method of claim 8, wherein the block copolymer is polystyrene-polymethylmethacrylate (PS-PMMA), polystyrene-polybutadiene (PS-PB), or polystyrene-polyisoprene (PS-PI). 9. The method of claim 8, wherein the mask metal is the same as the thin film metal on the substrate used in step (a). The method of claim 8, wherein step (d) comprises performing a UV etch followed by a reactive ion etch. 9. The method of claim 8, wherein step (f) uses reactive ion etching and ion milling. A nano-biochip manufactured by the method of any one of claims 1 to 7, wherein a bioreceptor that binds to or reacts with a target biomaterial is attached to a metal exposed by the nanopattern of the block copolymer. . 18. The nano-biochip according to claim 17, wherein the metal (M) is gold (Au). A bioreceptor BR prepared by the method of any one of claims 8 to 16 and attached to or reacted with a target biomaterial to a metal (M) nanopattern on a substrate (S) is attached. Nano-biochip of SM-BR type. 20. The nano-biochip according to claim 19, wherein the metal (M) is gold (Au). 19. A method for detecting a target biomaterial that binds to or reacts with a bioreceptor, using the nano-biochip of claim 18. 20. A method for detecting a target biomaterial that binds to or reacts with a bioreceptor, using the nano-biochip of claim 19.