KR101516322B1 - Magnetic nanowire coated with metal and method for preparing the same and biosensor for detecting biomolecule using the same - Google Patents
Magnetic nanowire coated with metal and method for preparing the same and biosensor for detecting biomolecule using the same Download PDFInfo
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- KR101516322B1 KR101516322B1 KR1020140012482A KR20140012482A KR101516322B1 KR 101516322 B1 KR101516322 B1 KR 101516322B1 KR 1020140012482 A KR1020140012482 A KR 1020140012482A KR 20140012482 A KR20140012482 A KR 20140012482A KR 101516322 B1 KR101516322 B1 KR 101516322B1
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- H01F1/00—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties
- H01F1/0036—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties showing low dimensional magnetism, i.e. spin rearrangements due to a restriction of dimensions, e.g. showing giant magnetoresistivity
- H01F1/0045—Zero dimensional, e.g. nanoparticles, soft nanoparticles for medical/biological use
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- C04B2237/00—Aspects relating to ceramic laminates or to joining of ceramic articles with other articles by heating
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Abstract
Description
The present invention relates to a metal-coated magnetic nanowire, a method of manufacturing the same, and a biosensor for detecting biomolecules using the same.
Nanoparticles called metal nanocomposites consist of spherical magnetic core nanoparticles and a thin metal film surrounding the core. The metal nanocomposite can absorb electromagnetic waves by the surface plasmon resonance phenomenon, and the absorbed electromagnetic waves collectively oscillate the conductive electrons in the metal film to generate heat. The wavelength range of the electromagnetic wave absorbed by the metal nanocomposite and the heat generated thereby can be easily controlled by adjusting the kind of the metal used and the thickness of the core nanoparticle and the metal film. Electromagnetic waves have a characteristic of being able to permeate a living tissue to a certain extent although there is a difference depending on the wavelength. Therefore, studies for applying the metal nanocomposite to drug delivery, diagnosis or treatment of diseases based on the characteristics of such metal nanocomposites and electromagnetic waves are continuously performed in the art.
One-dimensional nanostructures have attracted attention in recent years due to their unique properties and their role in making nanoscale devices (Ozin, 1992; Edelstein and Cammaratra, 1998; Xia et al., 2003). In addition, one-dimensional nanowires can be used as an important material for nanoscale electrical, optoelectronic, electrochemical, and electromechanical device applications. Currently, one-dimensional nanowires can be fabricated using electron beam or focused ion beam lithography (Gibson, 1997; Matsui and Ochiai, 1996), dip pen lithography (Piner et al., 1999), X-ray or polar-UV lithography (Tandon et al., 1991 ) Using a nano-lithography technique. However, since the nanolithography technique is complicated and not economical, a technique for manufacturing a one-dimensional nanowire more practically is required.
The field of molecular biology for observing and studying the constituents of plants and animals is based on the analysis of the characteristics and quantification of individual proteins or complex biological molecules, And is being applied to a wide range of medical fields.
A biosensor is a substance or device in which a biological substance such as a nucleic acid is immobilized on a substrate and a probe used for biochemical analysis is aligned with a biomolecule including DNA or the like. The principle of this biosensor is based on the interaction between the probe molecules immobilized on the substrate and the target molecules. Biosensors can be used not only to search for nucleic acids, proteins, and other substances that can bind nucleic acids, proteins, etc. immobilized on a substrate, but also to analyze whether target molecules bound to probe molecules exist in the sample It is possible. Thus, biosensors are widely used in biology research, medical diagnosis, drug discovery, and forensic science.
White salmon is a widely distributed salmon in the Pacific Rim and is a commercially valuable fishery resource. Recently, the white salmon ecosystem is seriously damaged by oil pollution, nuclear power plant accidents, and industrial wastewater pollution. Therefore, monitoring the DNA of white salmon is important as a tool to monitor the safety and marine pollution of seafood. Moriya et al., 2004; Moriya et al., 2007) have developed a DNA microarray for single-phase detection of mtDNA as a means to isolate the genetic basis of white salmon. However, this method has not been standardized due to the length problem of probe DNA, has poor reproducibility, and is not economical. As such, conventional biomolecule analytical methods are time-consuming, labor-intensive, and require expensive biomolecule analytical methods.
Therefore, it is urgently required to develop a biosensor capable of detecting biomolecules in a simple, economical and rapid manner.
The inventors of the present invention discovered that a biosensor using a metal-coated magnetic nanowire can detect various biomolecules in a simple and economical manner and quantitatively or qualitatively in a short time while searching for a biosensor for detecting biomolecules And completed the present invention.
Accordingly, the present invention provides a magnetic nanowire coated with a metal, a method for producing the same, and a biosensor for detecting biomolecules using the same.
In order to achieve the above object,
The present invention provides a magnetic nanowire having an average diameter of 15 to 25 nm and comprising metal-coated magnetic nanoparticles.
In addition,
Applying metal-coated magnetic nanoparticles to the top of the substrate; And
And aligning the magnetic nanoparticles by applying an external magnetic field to the substrate to which the magnetic nanoparticles are applied.
In addition,
A first electrode and a second electrode; A metal-coated magnetic nanowire connecting the first electrode and the second electrode; And a biomolecule-recognizing substance bonded to the metal-coated magnetic nanowire.
In addition,
1) fabricating a first electrode and a second electrode on a substrate;
2) applying metal-coated magnetic nanoparticles on the first and second electrodes;
3) applying an external magnetic field to the electrode to which the magnetic nanoparticles are applied, and connecting the first electrode and the second electrode to the magnetic nanowires coated with metal; And
4) binding the biomolecule-recognizing substance to the metal-coated magnetic nanowire.
The present invention also provides a biomolecule detection method comprising the step of measuring a change in current by applying a voltage between a first electrode and a second electrode of the biosensor.
In addition, the present invention provides a biomolecule detection kit including the biosensor.
Hereinafter, the present invention will be described in detail.
The present invention provides a magnetic nanowire comprising a magnetic nanoparticle coated with a metal having an average diameter of 15 to 25 nm.
In addition,
Applying metal-coated magnetic nanoparticles to the top of the substrate; And
And aligning the magnetic nanoparticles by applying an external magnetic field to the substrate to which the magnetic nanoparticles are applied.
The metal-coated magnetic nanoparticles may be prepared from a method known in the art or may be a commercially available material. For example, metal-coated magnetic nanoparticles can be prepared using a known method (Zhou et al., Sensors and Actuators B: Chemical 163, 224-232).
The metal-coated magnetic nanoparticles preferably have a core-shell structure composed of core particles responsive to magnetic force and metal particles coated around the core particles, but are not limited thereto.
The magnetic nanoparticles may be magnetic materials, magnetic alloys or magnetic oxides. Examples of the magnetic material include Co, Mn, Fe, Ni, Gd and Mo. Examples of the magnetic alloy include CoCu, CoPt, FePt, CoSm, NiFe and NiFeCo. Examples of the magnetic oxide include Fe 3 O 4 , Fe 2 O 3 , CoFe 2 O 4 , and MnFe 2 O 4 .
The magnetic nanoparticles are preferably selected from the group consisting of Fe 3 O 4 , Fe 2 O 3 , CoFe 2 O 4 , MnFe 2 O 4 , and combinations thereof, more preferably Fe 3 O 4 , But is not limited thereto.
The metal is preferably selected from the group consisting of gold (Au), silver (Ag), copper (Cu), platinum (Pt), nickel (Ni), aluminum (Al) Au) or silver (Ag), and gold (Au) is the most preferable, but is not limited thereto.
The metal may be produced by an electrolytic method or may be metal particles that are commonly used. In addition, metal precursors present in the solution can be reduced to metal to produce metal particles. Specifically, HAuCl 4 , HAuBr 4 , AgNO 3 and [Ag (NH 3 ) 2 ] NO 3 can be used as metal precursors for gold (Au) or silver (Ag)
The metal-coated magnetic nanoparticles preferably have an average diameter of 15 to 25 nm, more preferably 18 to 22 nm, but are not limited thereto.
The substrate is not particularly limited as long as it is generally used in this field. For example, glass, silicon, quartz, metal and polymer films (e.g., cycloolefin polymers, poly (alkyl (meth) acrylates), polystyrene, polyethylene, polypropylene, polyester, polyamino acid, Etc.), or a mixture of two or more kinds thereof. More preferably, a glass substrate can be used as the substrate, but is not limited thereto.
The external magnetic field is preferably generated by a permanent magnet such as ferrite, AlNiCo, neodymium (NdFeB), or samarium (SmCo), but is not limited thereto.
The direction in which the external magnetic field is applied is not particularly limited, and the permanent magnet may be placed on the upper or lower surface of the substrate.
The formation of the magnetic nanowires can be explained in two steps. First, the magnetic nanoparticles aggregate to form a chain, followed by chain and chain aggregation to form nanowires.
The length of the magnetic nanowires depends on the concentration of the magnetic nanoparticle solution. The larger the concentration of the solution, the longer and thicker nanowires can be obtained.
The magnetic nanowires preferably have an average length of 10 to 1000 μm, an average height of 100 to 150 nm, and an average width of 550 to 650 nm, but are not limited thereto.
According to one embodiment of the present invention, magnetic nanowires having an average length of 10 to 500 μm can be prepared when a metal-coated magnetic nanoparticle solution having a concentration of 1 to 15 μg / mL is added.
The present invention also provides a metal-coated magnetic nanowire produced by the above-described method.
In addition,
A first electrode and a second electrode; A metal-coated magnetic nanowire connecting the first electrode and the second electrode; And a biomolecule-recognizing substance bonded to the metal-coated magnetic nanowire.
The present invention also provides a method of manufacturing a semiconductor device, comprising: 1) fabricating a first electrode and a second electrode on a substrate; 2) applying metal-coated magnetic nanoparticles on the first and second electrodes; 3) applying an external magnetic field to the electrode to which the magnetic nanoparticles are applied, and connecting the first electrode and the second electrode to the magnetic nanowires coated with metal; And 4) binding the biomolecule-recognizing substance to the metal-coated magnetic nanowire.
The method of manufacturing a biosensor may further include a step of heat treating the first electrode and the second electrode, which are connected to each other by a magnetic nanowire coated with a metal, at 400 to 600 ° C in the presence of an inert gas.
Since the nanowires before the heat treatment exhibit low conductivity by interfering with the flow of electrons due to many voids existing on the surface, it is desirable to increase the conductivity of the nanowires through a heat treatment process. At this time, neon (Ne) or argon (Ar) may be used as the inert gas.
The electrode is preferably formed on a substrate such as glass, silicon, quartz or a polymer film and is preferably selected from the group consisting of gold, silver, platinum, nickel, cobalt, aluminum, copper, chromium, Electrode is more preferable, but it is not limited thereto.
In addition, the electrode may have various shapes such as a circle or a square, and the average length between the first electrode and the second electrode is preferably 5 to 20 μm, but is not limited thereto.
Detection and analysis of biomolecules is very important in biology research, medical diagnosis, drug discovery, and forensic medicine, such as searching for specific genes, sequencing, and determining whether viruses or pathogenic microorganisms are infected. Conventional biomolecule analytical methods are not only time consuming, but also require a lot of manpower and require expensive biomolecule analytical methods.
The biosensor of the present invention may include a biomolecule-recognizing substance bound to the metal-coated magnetic nanowires. In this case, the magnetic nanowire serves to connect the substrate with the biomolecule-recognizing substance.
The method of binding the biomolecule-recognizing substance to the metal-coated magnetic nanowire is not particularly limited. For example, a method of contacting the magnetic nanowire and the biomolecule-recognizing substance through an appropriate solvent such as PBS is used .
The biomolecule is a molecule that constitutes an organism and is a molecule necessary for the structure, function, and information transmission of an organism.
The biomolecule-recognizing substance means a biomolecule or other chemical substance capable of specifically binding with a biomolecule to be detected, and examples thereof include an antigen, an antibody, an RNA, a DNA, a hapten, an avidin Streptavidin, neutravidin, protein A, protein G, lectin, selectin, radioisotope labeling substance, aptamer and tumor marker, and the like), streptavidin, neutravidin, A substance capable of specifically binding, and the like, or a combination thereof.
When the tumor marker is an antigen, a substance capable of specifically binding to the antigen can be introduced into the biomolecule recognition substance, and examples thereof include a receptor or an antibody capable of specifically binding to the antigen. Examples of the antigen and the receptor or antibody capable of specifically binding thereto include EGF (epidermal growth factor) and anti-EGFR (ex. Cetuximab), C2 of synaptotagmin (C2 of synaptotagmin) and phosphatidylserine , Annexin V and phosphatidylserine, integrin and its receptor, VEGF (Vascular Endothelial Growth Factor) and its receptor, angiopoietin and Tie2 receptor, somatostatin and its receptor, HER2 / neu antigen (HER2 / neu antigen - breast cancer marker antigen) and Herceptin, prostate specific antigen (carcinoembryonic antigen - colorectal marker antigen) and Herceptin (Genentech, USA), vasointestinal peptide (Prostate-specific membrane antigen-prostate cancer marker antigen) and Rituxan (IDCE / Genentech, USA) and receptors thereof.
A representative example of a tumor marker " receptor " is a folic acid receptor that is expressed in ovarian cancer cells. A substance capable of specifically binding to the receptor (folate in the case of a folic acid receptor) can be introduced into the biosensor according to the present invention, and examples thereof include an antigen or an antibody capable of specifically binding to the receptor .
As described above, the antibody is a particularly preferred tissue-specific binding material in the present invention. In the present invention, the antibody includes polyclonal antibody, monoclonal antibody and antibody fragment. Antibodies have a property of selectively and stably binding only to a specific object, and -NH 2 of lysine in the Fc region of the antibody, -SH of cysteine, -COOH of aspartic acid and glutamic acid, And can be usefully used for bonding to nanowires.
Such antibodies are commercially available or can be prepared according to methods known in the art.
On the other hand, the above-mentioned " nucleic acid " includes RNA and DNA encoding the aforementioned antigen, receptor or a part thereof. Since a nucleic acid has a feature of forming a base pair between complementary sequences, a nucleic acid having a specific base sequence can be detected using a nucleic acid having a base sequence complementary to the base sequence. A nucleic acid having a nucleotide sequence complementary to the nucleic acid encoding the antigen or the receptor can be used in the biosensor according to the present invention.
The nucleic acid has a functional group such as -NH 2 , -SH or -COOH at the 5'- and 3'-terminal, and the functional group can be usefully used for binding nucleic acid to the magnetic nanowire of the present invention.
Such nucleic acids can be synthesized using standard methods known in the art, such as those available from automated DNA synthesizers (e.g., from Biosearch, Applied Biosystems, etc.)
In addition, the present invention provides a biosensor manufactured by the above production method.
The present invention also provides a biomolecule detection method comprising the step of measuring a change in current by applying a voltage between a first electrode and a second electrode of the biosensor. The type of the measuring device that can be included in the analyzer for detecting biomolecules is not particularly limited, and a general device known in this field can be used. For example, it is possible to use an apparatus capable of detecting an electric signal, a fluorescence signal or the like emitted from the biosensor, thereby realizing an electrical image or an optical image.
In addition, the present invention provides a biomolecule detection kit including the biosensor.
The biomolecule detection kit may include a biosensor capable of selectively recognizing biomolecules, as well as tools and reagents commonly used in the art used for immunological analysis. Examples of such tools and reagents include, but are not limited to, suitable carriers, labeling substances capable of producing a detectable signal, solubilizers, cleaning agents, buffers, stabilizers, and the like. When the labeling substance is an enzyme, it may include a substrate capable of measuring enzyme activity and a reaction terminator. Suitable carriers include, but are not limited to, soluble carriers, e. G., Physiologically acceptable buffers such as PBS, insoluble carriers such as polystyrene, polyethylene, polypropylene, Polyacrylonitrile, fluororesin, crosslinked dextran, polysaccharide, polymer such as magnetic fine particles plated with metal on latex, other paper, glass, metal, agarose and combinations thereof.
The biomolecule detection kit may be in the form of, but not limited to, an ELISA plate, a dip-stick device, an immunochromatographic test strip and a spin-split immune assay device, and a flow-through device .
Since the biosensor according to the present invention is manufactured using metal-coated magnetic nanowires, various biomolecules can be quantitatively or qualitatively detected simply, economically, and quickly.
FIG. 1 is a schematic diagram of a method of manufacturing a ferromagnetic nanowire including (a) a method of producing Fe 3 O 4 / Au nanoparticles, and Fe 3 O 4 / Au nanowires, (b) circular and rectangular electrodes, and (c) DNA.
2 is a transmission electron microscope (TEM) image and a size distribution bar graph (illustration) of (a) Fe 3 O 4 / Au nanoparticles according to Example 1 of the present invention, (b) (D) UV-Vis absorbance spectra and magnetization curves (not shown), and (c) Linear basic mapping and its mixture mapping, and (d) UV- Fig.
Fig. 3 is a graph showing the results when (a) 4 μg / mL of (b) 10 μg / mL of Fe 3 O 4 / Au nanoparticles were added, and ) Optical microscope image of the Fe 3 O 4 / Au nanowire before heat treatment produced on a square electrode.
4, 4 μg / mL Fe 3 O 4 / Au prior to the heat treatment prepared by adding the nanoparticles Fe 3 O 4 / Au (a ) AFM image of (b) cross-sectional height distribution of the nanowires according to the second embodiment of the present invention (C) AFM image of Fe 3 O 4 / Au nanowire connected between both electrodes, and (d) MFM image.
5 is a low magnification SEM image of (a) Fe 3 O 4 / Au nanowire before heat treatment prepared by adding 4 μg / mL Fe 3 O 4 / Au nanoparticles according to Example 2 of the present invention, (b) SEM image of the Fe 3 O 4 / Au nanowire connected between (c) the circular electrode and (d) the square electrode.
FIG. 6 is a graph showing current-voltage (IV) curves of Fe 3 O 4 / Au nanowires before and after annealing at 500 ° C. for 15 minutes under an argon atmosphere.
FIG. 7 shows (a) the IV curve and (b) the electrode attached to the DNA with no DNA, the electrode fixed with the probe DNA (HS-ss-DNA), and the target DNA according to Example 2 of the present invention, Fig. 7 is a diagram showing the magnitude of a detection signal.
Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.
Example 1. Preparation of metal-coated magnetic nanoparticles
1-1. Fe 3 O 4 Manufacturing
After completely dissolving 1.622 g of FeCl 3 .6H 2 O and 0.9941 g of FeCl 2 .4H 2 O in 40 mL of distilled water, 5 mL of ammonia solution (28 w / v%) was added to the mixed solution and stirred for 10 minutes Respectively. Then, 4.4 g of sodium citrate was added, the temperature was increased to 90 캜, and the mixture was stirred for 30 minutes. After completion of the reaction, the temperature was cooled to room temperature to obtain a black precipitate, which was washed with distilled water to prepare Fe 3 O 4 .
1-2. Fe 3 O 4 / Au nanoparticle production
20 mL of 0.5 mM HAuCl 4 solution was heated to boiling with stirring, and 10 mL of the Fe 3 O 4 solution prepared in the above 1-1 was added. Then, when the color of the mixed solution changed from brown to deep red, the heat source was removed, and the mixture was cooled to room temperature while stirring again. After completion of the reaction, the mixed solution was centrifuged at 5000 rpm for 30 minutes to remove Fe 3 O 4 particles, and the pure gold (Au) particles were separated by a permanent magnet. Thereafter, the solution containing the prepared Fe 3 O 4 / Au nanoparticles was stored at 4 ° C.
Example 2. Preparation of metal-coated magnetic nanowires
Magnetic plasmonic Fe 3 O 4 / Au nanowires were prepared from Fe 3 O 4 / Au nanoparticles using an external magnetic field. Specifically, the prepared Fe 3 O 4 / Au nanoparticles are aligned in the direction of the magnetic field using a neodymium magnet to obtain a one-dimensional Fe 3 O 4 / Au having an average height of 100-200 nm and a minimum length of 100 μm or more Nanowires were prepared. A method for producing the one-dimensional Fe 3 O 4 / Au nanowire is shown in FIG. .
2-1. Fabrication of gold electrode on glass substrate
As shown in FIG. 1 (b), a gold electrode having a distance of 10 μm (circular shape) and 15 μm (square shape) between electrodes is subjected to photolithography and lift-off processes Lt; / RTI >
First, a photoresist is spin-coated on a 4-inch glass substrate, and then UV light is selectively irradiated using a photomask and developed in an alkali solution to form a 1.4 μm thick mold pattern . Thereafter, chromium (Cr) and gold are sequentially deposited on a substrate having a mold pattern formed thereon by thermal evaporation, and then ultrasonic treatment is performed in an organic solvent to remove unnecessary metal patterns. Electrode.
2-2. Manufacture of magnetic nanowires
First, a gold electrode on the glass substrate prepared in Example 2-1 was placed between two magnets and 10 mm high from the magnets, and then a 4 to 10 μg / mL Fe 14 μL of 3 O 4 / Au nanoparticles were dropped onto the substrate. Then, the apparatus composed of the magnet and the electrode was evaporated by using an oven at 40 ° C. to produce a magnetic plasmonic Fe 3 O 4 / Au nanowire between the gold electrodes.
It was then heated to 500 캜 for 15 minutes under an argon atmosphere to improve the ohmic contact and conductivity between the magnetic plasmonic nanowire (MPNW) and the gold electrode.
Example 3. Fixation of probe DNA and detection of target DNA
3-1. Fixation of probe DNA
The gold electrode connected with the Fe 3 O 4 / Au nanowire prepared in Example 2 was ultrasonicated in acetone for 20 minutes, and then sufficiently washed with distilled water and methanol. The washed electrode was immersed in a PBS solution containing oligonucleotide (probe DNA) modified with 10 μM of 5'-alkanethiol, and then cultured for 1 hour, thereby forming an Fe 3 O 4 / Au nanowire The probe DNA-bound electrode was prepared.
3-2. Detection of target DNA
The probe DNA-bound electrode on the Fe 3 O 4 / Au nanowire prepared in Example 3-1 was immersed in a PBS solution containing 1 nM of target DNA and incubated for 1 hour to obtain probe DNA and target DNA Were complementarily combined. Then, the unbound target DNA was removed with PBS and distilled water, and the electrode to which the target DNA was bound was dried under a nitrogen atmosphere.
Experimental Example 1. Fe 3 O 4 / Au nanoparticle morphology, size, optical, and magnetic properties
(A) transmission electron microscope (TEM) images and size distribution histograms (diagrams) of Fe 3 O 4 / Au nanoparticles prepared according to Example 1 above, (b) elemental (d) UV-Vis absorbance spectrum and magnetization curves (illustration) are shown in Figure 2 (a) and Respectively. Here, the fundamental mapping in the direction of the line was measured by energy-dispersive X-ray spectroscopy line-scanning analysis of energy-dispersive X-ray spectroscopy, and the magnetization curve was measured using a superconducting quantum interference device (SQUID) .
As shown in FIG. 2 (a), the Fe 3 O 4 / Au nanoparticles of the present invention exhibited excellent dispersibility, high crystallinity, and uniform morphology, indicating that circular nanocrystals were generally formed. The size of the Fe 3 O 4 / Au nanoparticles was found to be about 20 nm in TEM, but was observed at about 25 nm when measured with a zeta-sizer. Thus, the difference in the measured values is due to the hydrodynamic size of the zeta cysteine nanoparticles. In addition, since the size of the Fe 3 O 4 / Au nanoparticles is about 10 nm (Zhou et al., Sensors and Actuators B: Chemical 163, 224-232), the thickness of the gold shell is estimated to be about 5 nm .
As shown in Figs. 2 (b) and 2 (c), the basic mapping for Fe (blue), O (red), and Au (green) of Fe 3 O 4 / Au nanoparticles is the spatial distribution of each element . According to these results, the gold (Au) shell showed the strongest and even mapping line. This is because the Au surface electron concentration is higher than that of Fe and O surface electrons. The Fe and O elements in the core showed weak signals due to low density surface electrons and gold shells. In addition, the linear basic mapping of each element displayed on the TEM was in good agreement with the Fe 3 O 4 / Au nanoparticles on the TEM, and the results showed that the nanoparticles were composed of gold (Au) shell and Fe 3 O 4 core .
As shown in Fig. 2 (d), the UV-Vis absorbance spectrum of Fe 3 O 4 nanoparticles showed no peak, but the absorption peak at 532 nm for Fe 3 O 4 / Au nanoparticles. This peak indicates the presence of a gold (Au) shell. This result was redshifted when compared to the absorption peak of 520 nm (J. Phys. Chem. B, 1999, 103, pp 4212-4217), which is an absorption peak of pure gold nanoparticles of similar size. The reason is thought to be due to the lack of electron density of the gold shell due to the uneven Au shell, and the interaction of the Fe 3 O 4 and the Au coating around the Fe 3 O 4.
Also, the saturation magnetization was decreased from 66.8 emu g -1 of Fe 3 O 4 nanoparticles with Fe 3 O 4 / Au 16.2 emu g -1 of the nanoparticles as shown in sapdo of 2 (d). This is because the magnetic cores are increased and the magnetic moment is decreased by coating the Fe 3 O 4 with gold (Au).
Experimental Example 2. Fe 3 O 4 / Au Analyzes the morphology, size, optical, and magnetic properties of nanowires
2-1. Fe before the heat treatment 3 O 4 / Au nanowire morphology analysis
(B) 10 μg / mL of Fe 3 O 4 / Au nanoparticles, (c) a circular electrode, and (d) a rectangular electrode according to Example 2, FIG. 3 shows an optical microscope image of the Fe 3 O 4 / Au nanowire before the heat treatment.
As shown in FIGS. 3 (a) and 3 (b), Fe 3 O 4 / Au nanoparticles at a concentration of 4 μg / mL produced nanowires with an average length of 70 μm, while Fe 3 O 4 / Au nanoparticles produced an average length of 350 μm nanowires. The results show that the larger the concentration of Fe 3 O 4 / Au nanoparticles, the longer and thicker the nanowires are formed.
As shown in FIGS. 3 (c) and 3 (d), it can be observed that the Fe 3 O 4 / Au nanowire is connected between the circular electrode and the rectangular electrode.
The formation of a magnetic plasmonic nanowire (MPNW) can be described in two steps. First, a chain of nanoparticles is formed (step 1), followed by chain-chain aggregation (step 2). The step (1) is formed by the Brownian motion, the steric effect, and the dipole interaction between the nanoparticles, and the step (2) is a step between the magnetic energy, Van der Waals attraction, surface energy, and entropy of the agglomerated chain It is caused by force. The main parameters of the chain structure depend on the nature of the particles (size, surfactant layer), particle concentration, and strength of the external magnetic field. Therefore, these variables must be combined well to obtain the desired structure.
(A) AFM image (b) cross-sectional height distribution diagram of the Fe 3 O 4 / Au nanowire before heat treatment prepared by adding 4 μg / mL Fe 3 O 4 / Au nanoparticles according to Example 2, (C) AFM image of the magnetic nanowire connected to the probe, and (d) MFM image is shown in FIG.
As shown in FIGS. 4 (a) and 4 (b), the Fe 3 O 4 / Au nanowire has a height of 130 nm and a width of 600 nm.
As shown in FIGS. 4 (c) and 4 (d), it can be observed that Fe 3 O 4 / Au nanowires are connected between the rectangular electrodes.
(A) low-magnification SEM image, (b) high magnification SEM image, and (b) high-resolution SEM image of the Fe 3 O 4 / Au nanowire before heat treatment prepared by adding 4 μg / mL Fe 3 O 4 / Au nanoparticles according to the above- (c) a circular electrode, and (d) a nanowire connected between the rectangular electrodes.
As shown in FIGS. 5 (a) and 5 (b), the SEM image of the Fe 3 O 4 / Au nanowire indicates the presence of many voids on the surface of the nanowire. This result means that the Fe 3 O 4 / Au nanowire is generated as a combination of Fe 3 O 4 / Au nanoparticles.
As shown in FIGS. 5 (c) and 5 (d), it can be observed that the Fe 3 O 4 / Au nanowire is connected between the circular electrode and the rectangular electrode.
2-2. Fe before and after the heat treatment 3 O 4 / Au Electron Characterization of Nanowires
Many voids present on the surface of Fe 3 O 4 / Au nanowires before heat treatment can interfere with the flow of surface electrons. Also, in a one-dimensional nanomaterial, the shape of the surface has a great influence on the overall conductivity. Thus, the Fe 3 O 4 / Au nanoparticles were heated to 500 ° C for 15 minutes under an argon atmosphere to form single metal nanowires. A current-voltage (IV) curve graph of the Fe 3 O 4 / Au nanowire after the heat treatment is shown in FIG.
As shown in FIG. 6, the Fe 3 O 4 / Au nanowires before annealing exhibited low conductivity and non-linearity. However, the Fe 3 O 4 / Au nanowires after heat treatment showed high conductivity and linearity. Conductivity increased about 11 times after heat treatment, compared to before heat treatment.
Experimental Example 3. Fixation of probe DNA and detection of target DNA
The heat-treated Fe 3 O 4 / Au nanowire according to Example 2 above was used for DNA detection biosensors. (A) IV curves and (b) the magnitude of the detection signals for the DNA-free electrode, the electrode bound to the probe DNA (HS-ss-DNA), and the electrode bound to the target DNA, respectively. Here, 10 μM probe DNA and 10 nM target DNA were bound to the Fe 3 O 4 / Au nanowire (( R - R 0 ) / R 0 ) to obtain the magnitude of the detection signal And R 0 is pure electrode resistance).
As shown in Fig. 7 (a), in the case of the probe DNA-bound electrode, the current decreased compared to the DNA-free electrode. This result indicates that the probe DNA is adsorbed on the Fe 3 O 4 / Au nanowire, and the surface charge-rich Fe 3 O 4 / Au nanowire is adsorbed to the external material to increase the resistance. In addition, the complementary binding of the probe DNA to the target DNA greatly increases the resistance, so that the electrode has the greatest reduction in current for the electrode bound to the target DNA.
As shown in FIG. 7 (b), the detection signal of the electrode to which the probe DNA was coupled showed a detection signal of 7.5%, and the detection signal of the electrode to which the target DNA was bound showed 27.5%.
Claims (19)
Wherein the magnetic nanoparticles are any one selected from the group consisting of Fe 3 O 4 , Fe 2 O 3 , CoFe 2 O 4 , MnFe 2 O 4 , CoPt, FePt, and combinations thereof.
Wherein the metal is any one selected from the group consisting of gold (Au), silver (Ag), copper (Cu), platinum (Pt), nickel (Ni) Nanowires.
And aligning the magnetic nanoparticles by applying an external magnetic field to the substrate to which the magnetic nanoparticles are applied.
Wherein the magnetic nanoparticles are any one selected from the group consisting of Fe 3 O 4 , Fe 2 O 3 , CoFe 2 O 4 , MnFe 2 O 4 , CoPt, FePt, and combinations thereof. Way.
Wherein the metal is any one selected from the group consisting of gold (Au), silver (Ag), copper (Cu), platinum (Pt), nickel (Ni) A method of manufacturing a nanowire.
Wherein the metal-coated magnetic nanoparticles have an average diameter of 15 to 25 nm.
Wherein the substrate is any one selected from the group consisting of glass, silicon, quartz, metal, polymer film, and combinations thereof.
Wherein the external magnetic field is generated by any one permanent magnet selected from the group consisting of ferrite, AlNiCo, neodymium (NdFeB), samarium (SmCo), and combinations thereof. ≪ / RTI >
Wherein the magnetic nanowires have an average length of 10 to 1000 μm, an average height of 100 to 150 nm, and an average width of 550 to 650 nm.
Wherein the magnetic nanowires have an average length of 10 to 500 mu m when a metal-coated magnetic nanoparticle solution having a concentration of 1 to 15 mu g / mL is added.
2) applying metal-coated magnetic nanoparticles on the first and second electrodes;
3) applying an external magnetic field to the first electrode and the second electrode to which the magnetic nanoparticles are applied, thereby connecting the first electrode and the second electrode to the magnetic nanowires coated with the metal; And
4) bonding the biomolecule-recognizing substance to the metal-coated magnetic nanowire.
Further comprising the step of heat-treating the first electrode and the second electrode connected to the metal-coated magnetic nanowire at 400 to 600 DEG C in the presence of an inert gas after the step 3).
Wherein the first electrode and the second electrode are any one selected from the group consisting of gold, silver, platinum, nickel, cobalt, aluminum, copper, chromium, and combinations thereof.
Wherein the first electrode and the second electrode have a circular or rectangular shape and an average length between the first electrode and the second electrode is 5 to 20 占 퐉.
The biomolecule-recognizing substance specifically binds to an antigen, an antibody, RNA, DNA, hapten, avidin, streptavidin, neutravidin, protein A, protein G, lectin, selectin, radioisotope labeling substance, Wherein the biomolecule is one selected from the group consisting of a substance capable of binding to the biological sample and a substance capable of binding to the biological sample.
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| KR20170119785A (en) * | 2016-04-19 | 2017-10-30 | 포항공과대학교 산학협력단 | Hybrid nanoparticles for detection of pathogenic bacteria and Manufacturing methods thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100802139B1 (en) * | 2006-08-08 | 2008-02-11 | 한국생명공학연구원 | Gold nanocages containing magnetic nanoparticles |
| KR20090000859A (en) * | 2007-06-28 | 2009-01-08 | 연세대학교 산학협력단 | A magnetic metal nano composite for the diagnosis and treatment |
| KR20110117138A (en) * | 2009-04-14 | 2011-10-26 | 서울대학교산학협력단 | Method for forming a microsphere having structural color |
| KR20130008395A (en) * | 2011-07-12 | 2013-01-22 | 아주대학교산학협력단 | Electrochemical biosensor for analyzing organophosphorus compounds and analytic method of organophosphorus compounds using it |
-
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- 2014-02-04 KR KR1020140012482A patent/KR101516322B1/en active IP Right Grant
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100802139B1 (en) * | 2006-08-08 | 2008-02-11 | 한국생명공학연구원 | Gold nanocages containing magnetic nanoparticles |
| KR20090000859A (en) * | 2007-06-28 | 2009-01-08 | 연세대학교 산학협력단 | A magnetic metal nano composite for the diagnosis and treatment |
| KR20110117138A (en) * | 2009-04-14 | 2011-10-26 | 서울대학교산학협력단 | Method for forming a microsphere having structural color |
| KR20130008395A (en) * | 2011-07-12 | 2013-01-22 | 아주대학교산학협력단 | Electrochemical biosensor for analyzing organophosphorus compounds and analytic method of organophosphorus compounds using it |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170119785A (en) * | 2016-04-19 | 2017-10-30 | 포항공과대학교 산학협력단 | Hybrid nanoparticles for detection of pathogenic bacteria and Manufacturing methods thereof |
| KR101871572B1 (en) | 2016-04-19 | 2018-06-27 | 포항공과대학교 산학협력단 | Hybrid nanoparticles for detection of pathogenic bacteria and Manufacturing methods thereof |
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