KR102426161B1 - Magnetic nanoparticles introduced with silane functional moiety - Google Patents

Abstract

In a method for measuring biomaterials such as DNA, RNA, and proteins in a living body, the present invention relates to magnetic nanoparticles, wherein when a silane functional group capable of specifically binding to the biomaterials is introduced into the magnetic nanoparticles and the magnetic nanoparticles are injected into the living body, the magnetic nanoparticles specifically bind to the biomaterials labeled with a fluorescent compound or the like to easily separate the same and efficiently detect the biomaterials.

Description

Magnetic nanoparticles introduced with silane functional moiety}

The present invention relates to a method for measuring a biomaterial such as DNA, RNA, or protein in a living body, when a functional group capable of specifically binding to the biomaterial is introduced into the magnetic nanoparticles, and the magnetic nanoparticles are injected into the living body. The present invention relates to magnetic nanoparticles capable of specifically binding to the biomaterial labeled with a fluorescent compound and the like, thereby easily separating the biomaterial, and efficiently detecting the biomaterial.

In general, a variety of nanoparticles can be used in order to measure the in vivo activity of a genetic material such as DNA, RNA, or a protein such as an antibody or to evaluate the effect. As nanoparticles, gold nanoparticles, semiconductor nanoparticles, magnetic nanoparticles, and the like are mainly used. The present invention relates to magnetic nanoparticles among the nanoparticles, and by introducing a silane functional group to the magnetic nanoparticles to specifically bind the biomaterials such as the dielectric material and protein, using a solvent capable of separating the magnetic nanoparticles. The present invention relates to magnetic nanoparticles capable of easily separating and detecting the biomaterial and the like.

Fluorescent and magnetic polymeric particles are known for their use as markers and indicators in a variety of biomedical assays. Among them, the most commonly used markers for cell sorting are, for example, immunoconjugates or immunological markers including immuno-fluorescence and immuno-magnetic markers. Immuno-fluorescent labels usually comprise, for example, a fluorescent molecule bound to an antibody. Immuno-magnetic labels usually comprise, for example, strong paramagnetic particles bound to a primary or secondary antibody. Cell labeling can be accomplished, for example, by attaching the antibody to a marker of interest (eg, a receptor site) on the surface of the cell, ie, a cell surface marker. However, the chemical and physical structures of cell surface markers and the density of immunological markers attached to the cell surface are generally difficult to accurately measure.

Magnetic particles, such as known magnetically active substances, can be bound or attached to antibodies, such as monoclonal antibodies, for example, with specificity for a particular cell type, antigen, or other target. The resulting magnetic-antibody can then be mixed with large populations of many individual cell types, such as, for example, crude tissue samples, cells grown in reactors, and the like. The magnetic-antibody thus attaches to the pre-selected target cell type, forming a magnetic-antibody-cell conjugate. The zygote can then be separated from the rest of the cell population using a magnetic field. Specific or non-specific binding of a cell or other biological target directly to a cell, for example by binding a specific paramagnetic compound to a specific hapten on the cell, or by a paramagnetic metal or metal complex to a cell, such as a metal-binding microorganism A disadvantage of magnetic particles is that they lack the specificity of magnetic labels, as they will be imparted paramagnetism by a number of distinct pathways that can confound analytical and diagnostic information generated by methods such as phagocytosis or by methods such as phagocytosis. . Other problems encountered with magnetic particles used in detection and diagnostics include, for example, the difficulty in obtaining highly accurate quantification of the magnetic susceptibility of a cell population.

Therefore, it is a very important problem to selectively attach magnetic nanoparticles to a biological material to be measured.

Efforts have been made to overcome the above problems by modifying the surface of magnetic nanoparticles with a silica compound. Silica is chemically stable, improves the dispersibility of nanoparticles, and has a high concentration of silanol groups on the surface, so it can be used for various surface bonding reactions and immobilization of other molecules. In a commonly used sol-gel process, it is easy to control the thickness of the silica layer formed on the outer shell of the magnetic nanoparticles by controlling the ammonia and TEOS/H ₂ O ratio. In addition, the silica shell not only protects the magnetic core, but also ensures dispersibility and oxidation stability in aqueous solution. However, when the thickness of the silica layer, which is a non-magnetic material, increases, the saturation magnetization of the magnetic nanoparticles decreases relatively rapidly.

When a large amount of magnetic nanoparticles are bonded to the outer shell of the silica core in a cluster form, the magnetic value (ie, saturation magnetization) and size can be controlled by controlling the size of the silica core and the number of magnetites bonded to the outer shell. Stoeva is J. Am. Chem. Soc., 127, 15362 (2005), a single silica core-clustered magnetite shell-type magnetic nanoparticle in which negatively-charged Fe ₃ O ₄ (15 nm) nanoparticles were bonded to the surface of an amine-functionalized silica core using electrostatic interaction. A complex was formed. Nagao reported that, in Langmuir, 24, 98049808 (2008), positively-charged magnetic nanoparticles (8 nm) were prepared and then mixed with silica nanoparticles (negatively-charged) prepared by Stmethod to obtain silica nanoparticles. A nanocomposite electrostatically bonded to the surface was prepared. Then, through the coating process of sodium silicate, a thin silica layer was formed on the outer shell to improve the stability of the magnetic nanoparticles. Salgueirino-Maceria Adv. Func. Mater., 15, 1036-1040 (2005), using LbL (layer-by-layer) technology to form an outer cluster layer by bonding cobalt/cobalt oxide nanoparticles to the surface of silica nanoparticles, and then coating them again with a silica layer did. The method for manufacturing a magnetic nanocomposite composed of such a single silica core-magnetite cluster shell is complicated in fixation, difficult to recover a large amount of magnetic nanoparticles not bound to the outer shell of the silica core, and oxidation stability of magnetic nanoparticles bound to the surface. And there is a problem that the formation of a secondary protective layer is required for surface functionalization.

Accordingly, the present inventors have developed magnetic nanoparticles into which a silane functional group capable of specifically binding to a desired biological material is introduced while maintaining the magnetization of the magnetic nanoparticles, and by coating the surface of the magnetic nanoparticles with silicon dioxide, The composition for labeling a biological material provided in the present invention containing magnetic nanoparticles prepared in this way has excellent sensitivity and affinity, and thus the binding efficiency with genetic material, protein, etc. in vivo is high. It is possible to improve the detection efficiency of biomaterials.

Korean Patent Publication No. 10-2011-0103009 Korean Patent Publication No. 10-2011-0035010

The present invention provides a biomaterial by introducing a silane functional group that allows the magnetic nanoparticles to specifically bind to a target biomaterial when measuring a biomaterial using a magnetic nanoparticle, and modifying the surface of the magnetic nanoparticle. It is intended to solve the problem of increasing specificity and sensitivity in labeling substances.

In addition, in the prior art, there was a problem that the process was complicated by independently applying magnetic nanoparticles, antibodies, and surfactants to conduct measurement experiments. It has the advantage of simplifying the measurement process.

In order to solve the above problems, the present invention solves the above problems by introducing a silane functional group that allows specific binding to biomaterials such as DNA, RNA, protein, and antibody to magnetic nanoparticles as shown in Chemical Formula 5 below. could solve it

<Formula 5>

ZnMnFe ₂ O ₄ @SiO ₂ @Ts

In the above formula, Ts represents a tosyl group represented by the following formula, @Ts is a magnetic particle coated with silicon dioxide (SiO ₂ ) ZnMnFe ₂ O ₄ @SiO ₂ Represented by the following formula 3 to which a tosyl group is bonded It indicates that it is coated with a compound that is

<Ts; Tosilgi>

<Formula 3>

The present invention can simplify the biomaterial measurement process using magnetic nanoparticles by introducing a silane functional group capable of specifically binding to biomaterials such as DNA, RNA, antibody, and protein on the surface of the magnetic nanoparticles, and also The specificity and sensitivity of the magnetic label can be improved by the nanoparticles being able to specifically bind to a desired biomaterial, and thus a high concentration of the conjugate in which the label compound into which the magnetic nanoparticles are introduced and the target material are bound can be obtained. Therefore, there is an advantage in that the reliability of the measurement result is remarkably superior to that of the prior art.

1 shows an electron micrograph of the magnetic nanoparticles prepared according to Example 1 of the present invention.
Figure 2 shows an electron micrograph of the magnetic nanoparticles prepared according to Example 2 of the present invention.
3 shows an electron micrograph of the magnetic nanoparticles prepared according to Example 3 of the present invention.
4 shows an electron micrograph of the magnetic nanoparticles prepared according to Example 4 of the present invention.
5 shows the results of analyzing the amount of streptavin labeled using the magnetic nanoparticles provided in the present invention and the amount of streptavin labeled using the conventional magnetic nanoparticles using an absorbance measuring device.

The present invention relates to magnetic nanoparticles used in combination with a compound that emits fluorescence, wherein the fluorescent material into which the magnetic nanoparticles are introduced binds very specifically to a biological material to be detected, such as DNA, RNA, protein, etc. , it is possible to obtain a high concentration of the magnetic nanoparticles and the fluorescent material conjugate to which the biomaterial is bound, so that the reliability of the result of detecting the biomaterial using fluorescence is very high.

The present inventors have completed the present invention by developing magnetic nanoparticles represented by the following Chemical Formula 5.

<Formula 5>

ZnMnFe ₂ O ₄ @SiO ₂ @Ts

In the above formula, Ts represents a tosyl group represented by the following formula, @Ts is a magnetic particle coated with silicon dioxide (SiO ₂ ) ZnMnFe ₂ O ₄ @SiO ₂ Represented by the following formula 3 to which a tosyl group is bonded It indicates that it is coated with a compound that is

<Ts; Tosilgi>

<Formula 3>

Hereinafter, a method for producing the magnetic nanoparticles represented by the above formula (5) will be described in detail.

The following Chemical Formulas 1 to 4 represent starting materials or intermediates for preparing Chemical Formula 5 above.

<Formula 1>

<Formula 2>

<Formula 3>

<Formula 4>

ZnMnFe ₂ O ₄ @SiO ₂

<Formula 5>

ZnMnFe ₂ O ₄ @SiO ₂ @Ts

In the above and below formulas, Ts represents a tosyl group represented by the following formula, and @Ts is a magnetic nanoparticle ZnMnFe ₂ O ₄ coated with silicon dioxide (SiO ₂ ) (Formula 4) The tosyl group below It represents a thing coated with a material represented by the following formula (3) that is bound (Formula 5).

<Ts; Tosilgi>

Synthesis Example 1. Preparation of a compound of the following formula (2)

8-nonene-1-ol (Formula 1, 10 g, 70.31 mmol) was dissolved in dichloromethane (200 mL), p -toluenesulfonyl chloride (16.08 g, 84.36 mmol), N,N-dimethyl-4-aminopyridine (860 mg) , 7.04 mmol) and pyridine (25.48 mL, 316.36 mmol) are added in this order.

After the reaction was stirred at room temperature for 4 hours, an aqueous hydrochloric acid solution (1 N, 300 mL) was added to terminate the reaction. The water layer and the organic layer are separated, and the water layer is extracted with dichloromethane. The organic layer was mixed, dried over MgSO ₄ , and distilled under reduced pressure, and the residue obtained was purified by flash chromatography (SiO ₂ ,40% EtOAc-hexanes) to obtain the compound of Formula 2 in the form of a yellow liquid (8.96 g).

¹ H NMR (500 MHz, DMSO) δ 7.78 (d, J = 10.0 Hz, 2H), 7.47 (d, J = 10.0 Hz, 2H), 5.83 - 5.74 (m, 1H), 4.99 - 4.91 (m, 2H) ), 4.00 (t, J = 6.4 Hz, 2H), 2.41 (s, 3H), 1.99 - 1.94 (m, 2H), 1.53 - 1.50 (m, 2H), 1.28 - 1.24 (m, 2H), 1.20 - 1.13 (m, 8H).

Synthesis Example 2. Preparation of a compound of Formula 3

DMF (100 mL) was added to a round-bottom flask dried by heating, and Chemical Formula 2 (4.36 g, 14.74 mmol) and Et ₃ OSiH (8.16 mL, 44.23 mmol) were added dropwise, and then ^karsdets Cat (20% Pt, 20% Pt, 1.76 mL, 1.48 mmol) is slowly added dropwise. Slowly increase the temperature from 0 ^o C to room temperature and stir for 4 hours. The solvent was removed by distillation under reduced pressure, and the residue was purified by flash chromatography (SiO ₂ ,30% EtOAc-hexanes) to obtain the compound of Formula 3 in the form of a pale yellow liquid (2.17 g).

¹ H NMR (500 MHz, DMSO) ¹ H NMR (500 MHz, DMSO) δ 7.78 (d, J = 5.0 Hz, 2H), 7.48 (d, J = 5.0 Hz, 2H), 4.05 - 3.98 (m, 2H) ), 3.75 - 3.68 (m, 6H), 2.42 (s, 1H), 1.54 - 1.51 (m, 2H), 1.30 - 1.04 (m, 25H)

Example 1. Preparation of a compound of Formula 5

Formula 4 (10 mL) was added to a 100 mL 1-neck round-bottom flask and stirred. A solution of Formula 3 (412 mg) and ethanol (4.8 mL) was slowly added dropwise to a 100 mL 1-neck round-bottom flask. After stirring for 10 minutes, a solution of ammonium hydroxide (1680 uL) and ethanol (4.16 mL) was slowly added dropwise.

After stirring the solution for 18 hours, it was put into a 50 mL nalgene tube and maintained on a magnetic stand for 10 seconds. After removing the magnetically separated filtrate, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 5 seconds.

After removing the magnetically separated ethanol, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 3 seconds. After the magnetically separated ethanol was removed and dried, a compound of Formula 5 was obtained.

Example 2. Preparation of the compound of formula 5

Formula 4 (10 mL) was added to a 100 mL 1-neck round-bottom flask and stirred. A solution of Formula 3 (412 mg) and ethanol (4.8 mL) was added drop-wise to a 100 mL 1-neck round-bottom flask. After stirring for 10 minutes, a solution of ammonium hydroxide (840 uL) and ethanol (4.16 mL) was slowly added dropwise.

After stirring the solution for 18 hours, it was put into a 50 mL nalgene tube and maintained on a magnetic stand for 10 seconds. After removing the magnetically separated filtrate, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 5 seconds. After removing the magnetically separated ethanol, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 3 seconds. Ethanol (10 mL) separated by a magnet was injected and dispersed.

The particles dispersed in ethanol (10 mL) were added to a 100 mL 1-neck round-bottom flask and stirred. A solution of Formula 3 (412 mg) and ethanol (4.8 mL) was slowly added dropwise to a 100 mL 1-neck round-bottom flask. After stirring for 10 minutes, a solution of ammonium hydroxide (840 uL) and ethanol (4.16 mL) was slowly added dropwise. After stirring for 18 hours, it was placed in a 50 mL nalgene tube and maintained on a magnetic stand for 10 seconds. After removing the magnetically separated filtrate, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 5 seconds. After removing the magnetically separated ethanol, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 3 seconds. After the magnetically separated ethanol was removed and dried, a compound of Formula 5 was obtained.

Example 3. Preparation of the compound of formula 5

Formula 4 (10 mL) was added to a 100 mL 1-neck round-bottom flask and stirred. A solution of Formula 3 (412 mg) and ethanol (4.8 mL) was slowly added dropwise to a 100 mL 1-neck round-bottom flask. After stirring for 10 minutes, a solution of ammonium hydroxide (840 uL) and ethanol (4.16 mL) was added drop-wise.

After stirring the solution for 18 hours, it was put into a 50 mL nalgene tube and maintained on a magnetic stand for 10 seconds. After removing the magnetically separated filtrate, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 5 seconds. After removing the magnetically separated ethanol, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 3 seconds. After the magnetically separated ethanol was removed and dried, a compound of Formula 5 was obtained.

Example 4. Preparation of the compound of formula 5

Formula 4 (10 mL) was added to a 100 mL 1-neck round-bottom flask and stirred. A solution of Formula 3 (206 mg), tetraethyl orthosilicate (100 uL), and ethanol (4.8 mL) was slowly added dropwise to a 100 mL 1-neck round-bottom flask. After stirring for 10 minutes, a solution of ammonium hydroxide (840 uL) and ethanol (4.16 mL) was slowly added dropwise.

After stirring the solution for 18 hours, it was put into a 50 mL nalgene tube and maintained on a magnetic stand for 10 seconds. After removing the magnetically separated filtrate, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 5 seconds. After removing the magnetically separated ethanol, fresh ethanol (30 mL) was injected and dispersed. Place a 50 mL tube on a magnetic stand, add a solution dispersed in ethanol, and hold for 3 seconds. After the magnetically separated ethanol was removed and dried, a compound of Formula 5 was obtained.

Another object to be provided by the present invention relates to a method for detecting a biological material present in a sample using the magnetic nanoparticles represented by the formula (5), wherein the detection method is represented by the following formula (5) in the sample A first step of mixing magnetic nanoparticles and a fluorescent compound to perform a labeling reaction on the biological material;

a second step of obtaining magnetic nanoparticles by first filtration after performing the labeling reaction and standing on a magnetic stand;

A third step of obtaining magnetic nanoparticles after a second filtration after adding a buffer solution to the magnetic nanoparticles obtained in the second step, allowing a blocking reaction, and then standing on a magnetic stand;

Measuring the absorbance of the biomaterial labeled on the magnetic nanoparticles obtained in the third step; characterized in that it consists of.

The biomaterial is mainly DNA, RNA and protein, and the fluorescent compound is not limited to a specific fluorescent compound, and any fluorescent compound for labeling the biomaterial may be used.

Hereinafter, the magnetic nanoparticles provided in the present invention are further compared with the conventional magnetic nanoparticles by comparing the results applied to the detection of the physical properties and biomaterials of the magnetic nanoparticles represented by Chemical Formula 5 prepared through the above Examples. Describe in detail. However, the examples are only intended to illustrate the present invention, and are not intended to limit the scope of the present invention.

Test Example 1. Measurement of the size of the magnetic nanoparticles of Formula 5 provided in the present invention

In order to confirm the size of the magnetic nanoparticles according to the present invention, transmission electron microscopy was performed. The compounds of Examples 1 to 4 were dispersed by sonication, and 5 uL was put on a carbon grid and dried. The size of the compound was confirmed using a transmission electron microscope, and the results are presented in FIGS. 1 to 4 .

Test Example 2. Confirmation of the performance of the functional group of the magnetic nanoparticles of Formula 5 provided in the present invention

After labeling the protein to confirm the performance of the functional group of the magnetic nanoparticles introduced in the present invention, the amount of the labeled protein was measured. The protein used was Streptavidin, and the number of samples in each experiment was carried out by three.

The magnetic nanoparticles are dispersed in 0.1 M sodium borate buffer (pH 9.5) and adjusted to a concentration of 10 mg/mL. Dispense 200 uL (2 mg) of dispersed magnetic nanoparticles into a 1.5 mL ep tube, place it on a magnetic stand, and remove the clear filtrate except for the magnetic nanoparticles. After injecting 40 uL of 0.1M sodium borate buffer (pH 9.5) into the magnetic nanoparticles from which the filtrate has been removed, disperse them, put them on a magnetic stand again, and remove the filtrate to complete the washing process of the magnetic nanoparticles.

Before proceeding with the labeling reaction, Streptavidin is dispersed in 0.1M sodium borate buffer (pH 9.5) in advance and adjusted to a concentration of 10 mg/mL.

After the washing is completed, 13.4 uL of 0.1M sodium borate buffer (pH 9.5) is injected into the magnetic nanoparticles and dispersed. After treating the dispersed magnetic nanoparticle mixture with 20 uL (200 ug) of Streptavidin, vortex lightly.

After treating the mixture with 16.6 uL of 3M ammonium sulphate solution to adjust the total volume of the mixture to 50 uL, the mixture was incubated in a Multi-mixer (b3 mode, 5 rpm), 37°C, for 24 hours for labeling reaction. proceed with

After the reaction was completed, the mixture was left on a magnetic stand for 2 minutes, and then 50 uL of the first filtrate was obtained (Filtrate No. 1).

After the filtrate is removed, add 50 uL of 0.01M phosphate buffered saline (pH 7.4, with 0.5% BSA and 0.05% Tween®20) to the remaining magnetic nanoparticles and vortex lightly.

The mixture was reacted in an incubator Multi-mixer (b3 mode, 5 rpm, 37° C.) for 24 hours to perform a blocking reaction. After the reaction mixture was left on a magnetic stand for 2 minutes, 50 uL of a second filtrate was obtained (Filtrate No. 2).

The amount of streptavidin labeled on magnetic nanoparticles was measured from filtrate No. 1 and filtrate No. 2 obtained in the above experiment, and the amount was analyzed using absorbance measuring equipment NanoDrop™ 2000 Spectrophotometer. Since the amount of the sample to be measured is not large, it was selected as the absorbance measuring device, and the measured value was performed by increasing the number of n samples (n=3) to compensate for the error range while obtaining the average value.

The absorbance measurement process is as follows. First, draw a standard deviation line at the 280 nm wavelength of streptavidin. Since the buffers of Filtrate 1 and Filtrate 2 are different, prepare two standard streptavidin solutions in which filtrates 1 and 2 are respectively dissolved in the same buffer. Using NanoDrop™ equipment, dilute the streptavidin standard solution by 1/2 and measure the absorbance at each 280 nm wavelength. After deriving two standard deviation lines using the obtained absorbance data, a trend line equation is obtained.

After measuring the absorbance of the filtrate No. 1 at 280 nm wavelength, the average amount of streptavidin (ug) in the filtrate No. 1 is obtained by substituting it into the corresponding streptavidin standard deviation trend line equation. The average amount (ug) of streptavidin labeled on magnetic nanoparticles is obtained by subtracting the initial amount of streptavidin (200 ug) from the average amount of streptavidin (ug) obtained above. Absorbance is measured in the same manner for filtrate No. 2, but only the average amount of streptavidin (ug) in the filtrate is calculated. This is to measure the amount of streptavidin separated from magnetic nanoparticles in the blocking step. Finally, the average amount (ug) of streptavidin labeled on the magnetic nanoparticles provided in the present invention is [average amount of labeled streptavidin obtained through filtrate 1 (ug) - average amount of streptavidin in filtrate 2 (ug)] to be.

For a more accurate comparison, ThermoFisher scientific's Dynabeads™ MyOne™ Tosylactivated (catalog 65501) product, which is a conventional magnetic nanoparticle, was selected as a reference. The results of the previously obtained average amount of streptavidin (ug) and the streptavidin labeling rate (%) compared to the conventional magnetic nanoparticles are presented in Tables 1 and 2 and FIG. 5 .

Sample Dynabeads™ MyOne™ Tosylactivated Compound of Formula 5 (Example 1) Compound of Formula 5 (Example 2) Streptavidin
Average labeling amount (ug) 35.3 40.7 31.7

Sample Dynabeads™ MyOne™ Tosylactivated Compound of Formula 5 (Example 1) Compound of Formula 5 (Example 2) Third-party magnetic nanoparticles
prepare (%) 100 115 90

As shown in Tables 1 and 2 and FIG. 5, the absorbance of the protein was compared with the result of measuring the amount of Streptavidin using the magnetic nanoparticles provided in the present invention and the magnetic nanoparticles provided in the prior art. As a result, when the magnetic nanoparticles provided in the present invention were used and absorbance was measured through two filtration steps, much superior results were obtained than in the prior art.

The present invention is industrially applicable.

Claims (8)

Magnetic nanoparticles for labeling biomaterials represented by the following Chemical Formula 5
<Formula 5>
ZnMnFe ₂ O ₄ @SiO ₂ @Ts
In the above formula, Ts represents a tosyl group represented by the following formula, @Ts is a magnetic particle coated with silicon dioxide (SiO ₂ ) ZnMnFe ₂ O ₄ @SiO ₂ Represented by the following formula 3 to which a tosyl group is bonded It indicates that it is coated with a compound that is
<Ts;Tosilgi>

<Formula 3>

As a method for producing the magnetic nanoparticles of claim 1,
A first step of preparing a compound represented by the following Chemical Formula 1 and p-toluenesulfonyl chloride, NN-dimethyl-4-aminopyridine and pyridine mixed in a dichloromethane solvent to prepare a compound represented by the following Chemical Formula 2;
a second step of preparing a compound represented by the following formula (3) by dropwise adding triethoxysilane to the composition containing the compound of formula (2) obtained in the first step;
A mixture of the compound represented by Formula 3 and ethanol obtained in the second step was added dropwise to the compound represented by Formula 4 below, and then a mixture of ammonium hydroxide and ethanol was added dropwise to the compound represented by Formula 5 below. the third step of manufacturing;
Method for producing magnetic nanoparticles comprising the
<Formula 1>

<Formula 2>

<Formula 3>

<Formula 4>
ZnMnFe ₂ O ₄ @SiO ₂
<Formula 5>
ZnMnFe ₂ O ₄ @SiO ₂ @Ts
In the above and below formulas, Ts represents a tosyl group represented by the following formula, @Ts is a magnetic particle of ZnMnFe ₂ O ₄ coated with silicon dioxide (SiO ₂ ) (Formula 4) and the silicon dioxide The coated magnetic particles of ZnMnFe ₂ O ₄ @SiO ₂ (Formula 4) are coated with a compound represented by Formula 3 having a tosyl group (Formula 5).
<Ts;Tosilgi>

The method according to claim 2, wherein tetraethyl orthosilicate is further added dropwise to a mixture of the compound represented by Formula 3 and ethanol added dropwise in the third step of the manufacturing method.
The magnetic nanoparticles according to claim 1, wherein the biomaterial is any one selected from DNA, RNA, and protein.
Composition for labeling biomaterials formed including magnetic nanoparticles represented by the following Chemical Formula 5
<Formula 5>
ZnMnFe ₂ O ₄ @SiO ₂ @Ts
In the above formula, Ts represents a tosyl group represented by the following formula, @Ts is a magnetic particle coated with silicon dioxide (SiO ₂ ) ZnMnFe ₂ O ₄ @SiO ₂ Represented by the following formula 3 to which a tosyl group is bonded It indicates that it is coated with a compound that is
<Ts;Tosilgi>

<Formula 3>

The composition for labeling a biological material according to claim 5, wherein the biological material is any one selected from DNA, RNA, and protein.
A method for detecting a biological material in a sample, comprising:
The detection method may include a first step of mixing a sample with magnetic nanoparticles represented by the following Chemical Formula 5 and a fluorescent compound to perform a labeling reaction on the biological material;
a second step of obtaining magnetic nanoparticles by first filtration after performing the labeling reaction and then standing on a magnetic stand;
A third step of obtaining magnetic nanoparticles after a second filtration after adding a buffer solution to the magnetic nanoparticles obtained in the second step, allowing a blocking reaction, and then standing on a magnetic stand;
Measuring the absorbance of the biological material labeled on the magnetic nanoparticles obtained in the third step; Method for detecting a biological material, characterized in that consisting of
<Formula 5>
ZnMnFe ₂ O ₄ @SiO ₂ @Ts
In the above formula, Ts represents a tosyl group represented by the following formula, @Ts is a magnetic particle coated with silicon dioxide (SiO ₂ ) ZnMnFe ₂ O ₄ @SiO ₂ Represented by the following formula 3 to which a tosyl group is bonded It indicates that it is coated with a compound that is
<Ts;Tosilgi>

<Formula 3>

The method according to claim 7, wherein the biomaterial is any one selected from DNA, RNA, and protein.

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