KR20220118336A - Nanoparticle complex coated with macromolecule for isolating biological target and preparation method the same - Google Patents

Abstract

The present invention relates to a nanoparticle complex for isolating a biological target, and a preparation method thereof, and specifically relates to a nanoparticle complex in which a receptor is bound to nanoparticles coated with macromolecule, a preparation method thereof, and a use thereof. The macromolecule is one or more selected from a group including polydopamine, polyethylene glycol, polyetherimide, polyvinyl alcohol, casein, dextran, and chitosan. According to the present invention, the nanoparticle complex can be effectively used for diagnosis of various diseases.

Description

Nanoparticle complex coated with macromolecule for isolating biological target and preparation method the same}

The present invention relates to a nanoparticle complex for separation of a biological target, a method for manufacturing the same, and more specifically, a nanoparticle complex in which a receptor is bound to a nanoparticle coated with a polymer, and the matrix effect in a biological sample in which various components are present in complex It relates to a nanoparticle complex that can effectively maintain a dispersed state of the nanoparticle complex by reducing

At this point in time when interest in health and diseases is increasing, research on methods for managing health and reducing treatment costs through early diagnosis of diseases are being actively conducted. Among them, there have been many attempts to detect diseases by recognizing or isolating specific biomarkers in biological samples in which various components exist in a complex manner using nanoparticles, but proteins in biological samples such as blood, saliva, and urine Since a large amount of other impurities, including cellular substances in the form of , nucleic acids, and carbohydrates, exist, aggregation between particles occurs due to the matrix effect of nanoparticles during disease diagnosis, thereby significantly improving the detection sensitivity of the target substance. Not only is it reduced, but it is difficult to isolate the target material efficiently.

Therefore, if particles can be efficiently dispersed by preventing aggregation of particles by preventing a matrix effect in a biological sample, not only can the detection sensitivity be remarkably increased by increasing the binding force to the target, but also the target material can be separated with high efficiency and/or detectable.

Domestic registered patent 10-1646610

The present invention has been devised to solve the problems in the prior art as described above, and relates to a nanoparticle complex in which a receptor is bound to nanoparticles coated with a polymer, and the dispersing power of nanoparticles by inhibiting the matrix effect by coating the nanoparticles It is an object of the present invention to provide a nanoparticle complex, a method for preparing the same, and a use thereof, which have significantly increased detection sensitivity through this.

However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned can be clearly understood by those of ordinary skill in the art to which the present invention belongs from the following description. will be.

The present invention provides a nanoparticle complex for detecting or separating a target material from a biological sample. In the nanoparticle complex, the surface of the nanoparticles is preferably coated with a macromolecule to prevent a matrix effect in a biological sample, and the polymeric material includes a receptor for detecting or separating the target material. may be bound to specifically bind to a target substance. And the polymer material is preferably polydopamine, polyethylene glycol, polyetherimide, polyvinyl alcohol, casein, dextran, and chitosan ( chitosan) may be any one or more selected from the group consisting of, more preferably polydopamine or polyethylene glycol, or a polymer material that can be coated on nanoparticles by self-polymerization, or can bind to nanoparticles having a thiol group If so, it is not limited thereto. In addition, the coating may be preferably coated in a thickness of 1 to 500 nm, more preferably in a thickness of 1 to 100 nm, more preferably in a thickness of 1 to 30 nm, but may prevent matrix effect. The thickness is not limited thereto.

In one embodiment of the present invention, the coating may be coated by mixing a monosaccharide for binding to lectin and the like with a polymer material, and the monosaccharide includes mannose, glucose, fructose. ), glucosamine, and the like, but is not limited thereto as long as it is a monosaccharide capable of binding to lectin by electrostatic bonding.

In another embodiment of the present invention, the bond is any one or more types of bonds selected from the group consisting of an ionic bond, a covalent bond, a coordination bond, a hydrogen bond, a hydrophobic interaction, a van der Waals interaction, and a bond by a linker. It may be, but is not limited thereto, as long as it is a form known as a method for binding a polymer material and a receptor.

In another embodiment of the present invention, the bonding by the linker is characterized in that the bonding by a 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) linker or a thiol group.

In another embodiment of the present invention, the receptor may be an antibiotic, antibody, nucleic acid, protein, lectin, etc. that specifically binds to a target substance, and the nucleic acid is plasmid DNA, cDNA, siRNA, miRNA, antisense. RNA, aptamers, oligonucleotides, and the like. However, if it is a receptor capable of specifically binding to a target material to detect or isolate the target material, the present invention is not limited thereto.

In another embodiment of the present invention, the target material may be a pathogen, a virus, a cell, a protein, a nucleic acid, an antigen, a glycoprotein, a metal, etc., as long as it is a biomarker that can be used to diagnose a disease, but is not limited thereto.

In another embodiment of the present invention, the nanoparticles may be silica nanoparticles, iron oxide nanoparticles, polystyrene nanoparticles, and the like, but is not limited thereto as long as the nanoparticles can be coated with a polymer material.

In another embodiment of the present invention, the diameter of the nanoparticle composite may be preferably 200 to 1,000 nm, more preferably, 200 to 800 nm, but is not limited thereto.

In another embodiment of the present invention, the biological sample is preferably blood, saliva, bone marrow fluid, lymph fluid, urine, amniotic fluid, mucosal fluid, peritoneal fluid, etc., but cells, nucleic acids, proteins, extracellular vesicles, impurities If it is a biological sample in which various substances are complexly mixed, the present invention is not limited thereto.

The present invention also provides a method for preparing a nanoparticle composite comprising the following steps: a) preparing polydopamine-coated nanoparticles by adding and dispersing nanoparticles and an acid-base catalyst to a polydopamine solution ; and b) adding a receptor to the polydopamine-coated nanoparticles and reacting to prepare a polydopamine-coated nanoparticle complex to which the receptor is bound.

In one embodiment of the present invention, the nanoparticles and polydopamine of step a) may be mixed in a weight ratio of 1: 0.01 to 1:1, and monosaccharides are added to the polydopamine solution of step a) from 1: 0.5 to It may be additionally added in a weight ratio of 1:1.5.

In another embodiment of the present invention, the nanoparticles may be preferably iron oxide nanoparticles, polystyrene nanoparticles, and the like, but is not limited thereto as long as it is a type of nanoparticles that can be coated by a self-polymerization reaction with a polymer material.

In another embodiment of the present invention, step a) may be dispersed for 4 hours to 36 hours, and step b) may be reacted for 1 hour to 5 hours.

The present invention also provides a method for producing a nanoparticle composite comprising the steps of: a) preparing nanoparticles coated with a polymer material by adding and dispersing a polymer material to silica shell nanoparticles having a thiol group; and b) adding and reacting a receptor to the coated nanoparticles to prepare a nanoparticle composite coated with a polymer material to which the receptor is bound, wherein the polymer material is polyethylene glycol, poly It may be etherimide (Polyetherimide), polyvinyl alcohol (polyvinyl alcohol), casein (casein), dextran (dextran), chitosan (chitosan), and the like.

In one embodiment of the present invention, step b) may be reacted by adding 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) together with the receptor.

In another embodiment of the present invention, the nanoparticles of step a) and polyethylene glycol or polyvinyl alcohol may be mixed in a weight ratio of 1:1 to 1:5 and dispersed for 4 hours to 36 hours.

In another embodiment of the present invention, step b) may be reacted for 1 hour to 5 hours.

In addition, the present invention provides a method for detecting or separating a target material from a biological sample using the nanoparticle complex. The method is characterized in that it comprises the step of treating and reacting the nanoparticle complex to the biological sample in vitro.

In addition, the present invention provides a method for diagnosing an infectious disease or a method for providing information on the diagnosis of an infectious disease using the nanoparticle complex. The method is characterized in that it comprises the step of treating and reacting the nanoparticle complex to the biological sample in vitro.

In addition, the present invention provides a use for detecting or separating a target material from a biological sample of the nanoparticle complex.

In addition, the present invention provides the use of the nanoparticle complex for diagnosis of infectious diseases.

In addition, the present invention provides a kit for detecting or separating a target material from a biological sample, comprising the nanoparticle complex as an active ingredient.

In addition, the present invention provides a kit for diagnosing an infection comprising the nanoparticle complex as an active ingredient.

The nanoparticle complex according to the present invention uses nanoparticles coated with a polymer, so that various receptors can be effectively bound to the nanoparticles to easily produce a nanoparticle complex, and through this, not only infectious diseases caused by pathogens, viruses, but also cancer It can be effectively used for diagnosis of various diseases using biomarkers such as In addition, aggregation between particles does not occur even when the nanoparticle complex is directly treated on a biological sample without pre-treatment of the biological sample by preventing the matrix effect occurring in the biological sample by using the nanoparticles coated with the polymer. Through this, the detection sensitivity can be significantly increased by preventing fouling. Therefore, the nanoparticle complex of the present invention is expected to be widely used in the separation, detection, diagnosis, treatment, etc. of various biological targets using receptors.

1 is a view showing the results of observing nanoparticles coated with a polymer material according to an embodiment of the present invention with a scanning electron microscope.
2 is a view showing the results of observing nanoparticles coated with a polymer material according to an embodiment of the present invention with a transmission electron microscope.
3 is a diagram schematically illustrating a method for preparing a PDA-coated nanoparticle complex bound to an antibody or antibiotic according to an embodiment of the present invention and a binding mechanism with a target.
4 is a view briefly showing a method of manufacturing a lectin-coupled Man/PDA-coated nanoparticle complex and a binding mechanism with a target according to an embodiment of the present invention.
5 is a diagram schematically illustrating a method for preparing a PEG-coated nanoparticle complex to which an antibody or antibiotic is bound according to an embodiment of the present invention.
6 is a view showing the results of confirming the dispersibility of nanoparticles according to an embodiment of the present invention.
7 is a view showing the results of confirming the resolution of the target using the nanoparticle complex coated with the polymer material to which the antibiotic is bound according to an embodiment of the present invention.

The nanoparticle composite of the present invention is a nanoparticle composite in which a receptor is bound to a nanoparticle coated with a polymer, and a polymer such as PDA, PEG, or PVA is coated on the surface of the nanoparticle in a nano-thickness to prevent the matrix effect in the biological sample By doing so, the detection sensitivity of the target material contained in the biological sample can be significantly improved, and even if a small amount of the target material is included, it can be detected and/or separated with high accuracy, so that it can be effectively used for diagnosis of various diseases.

As used herein, "specifically binding" refers to binding to a target material to be separated or detected by specifically interacting with the receptor and target material bound to the nanoparticle complex of the present invention. It means the form that is combined. The target material may include all substances or biomarkers generally used for diagnosing diseases, such as nucleic acids, antigens, proteins, cells, cancer cells, pathogens, viruses, and metals, included in a biological sample.

As used herein, the term “receptor” refers to a substance that specifically binds to the target substance, and preferably an antibiotic, an antibody, a nucleic acid, a protein, a lectin, etc., or a target substance that specifically binds to the target substance If the substance can be detected or separated, it is not limited thereto.

As used herein, the term “nanoparticle” includes all commonly used nano-sized particles, preferably silicon particles, polystyrene particles, latex particles, metal particles, glass particles, magnetic particles, silica shell nanoparticles. It may be selected from the group consisting of particles and combinations thereof, but is not limited thereto.

As used herein, the term “biological sample” may be any biological sample in which the target cells may be present, for example, a biopsy sample, a tissue sample, or a cell suspension in which isolated cells are suspended in a liquid medium. , cell culture, body fluid, and combinations thereof, and the body fluid may include blood, saliva, bone marrow fluid, lymph fluid, urine, amniotic fluid, mucosal fluid, peritoneal fluid, and the like.

As used herein, the term "coating" refers to coating a thin film using a polymer material on the surface of nanoparticles, and the thin film is 1 to 500 nm, 1 to 200 nm, 1 to 100 nm, 1 to 50 nm, Or it may have a thickness of 1 to 30 nm, but is not limited thereto.

As used herein, the term "kit" refers to a device for detecting or separating a target material from a biological sample, including the nanoparticle complex of the present invention as an active ingredient, and a target material from a biological sample separated from an organism There is no limitation as long as it is a form in which the presence or absence of the target substance or the amount of the target substance can be confirmed. The kit can be used in various diagnostic fields by checking the presence or amount of a target material bound to the nanoparticle complex of the present invention, and preferably can also be used for diagnosing an infection.

As used herein, the term “infection disease” refers to any disease resulting from infection of a pathogenic microorganism, and the pathogenic microorganism may be bacteria, viruses, protozoa, fungi, etc. Any microorganism known to be inducible is not limited thereto.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1: Preparation of nanoparticles coated with a polymer material

In order to prepare nanoparticles for preventing a matrix effect, which is a problem when analyzing a biological sample in which various components are complexly mixed, nanoparticles coated with a polymer material were prepared. In order to select the polymer material that can most effectively prevent the matrix effect, polydopamine (PDA), polyethylene glycol (PEG), polyetherimide (PEI), polyvinyl alcohol (PVA) ), casein, dextran, and chitosan were used to prepare nanoparticles coated with a polymer material.

The iron oxide nanoparticles were prepared by a generally known method. More specifically, 100 mg of Fe ₃ O ₄ powder (100 nm) was added to 40 mL of 1M HCl (hydrochloric acid) solution and mixed, followed by reaction at room temperature for 1 hour. After collecting the particles through a permanent magnet, the remaining solution is removed, the collected particles are added to 40 mL of phosphate buffered saline (PBS, pH 7.4), vortexed for 1 minute, and again using a permanent magnet. It was washed three times in such a way that only particles were collected. And finally, the prepared iron oxide nanoparticles were stored in 10 mL of phosphate buffer solution.

And to prepare PDA-coated nanoparticles, firstly, 50 mg of dopamine is added to 25 mL of Tris-HCl buffer (pH 8.5) or acetic acid-added buffer as an acid-base catalyst to prepare a PDA solution, Then, nanoparticles such as iron oxide nanoparticles or polystyrene nanoparticles were added, dispersed by ultrasonication for 16 hours, and PDA-coated nanoparticles (NPs@PDA) were prepared through a self-polymerization reaction. . After collecting the particles through a permanent magnet, the remaining solution is removed, the collected particles are added to 40 mL of phosphate buffered saline (PBS, pH 7.4), vortexed for 1 minute, and again using a permanent magnet. It was washed three times in such a way that only particles were collected. And finally, the prepared PDA-coated nanoparticles were stored in 1 mL of phosphate buffer solution. At this time, the content ratio of nanoparticles and PDA was variously changed, and PDA-coated nanoparticles were prepared. When the content ratio of nanoparticles and PDA was 1: 0.01 to 1: 1.5 by weight, PDA-coated nanoparticles were effectively produced was prepared, and when the weight ratio was greater than 1: 1.5, the nanoparticles were agglomerated with each other, and when the weight ratio was less than 1: 0.01, it was confirmed that the PDA coating was not sufficient. In addition, it was confirmed that the coating thickness of the PDA decreased as the self-polymerization reaction time was decreased, and the coating thickness of the PDA was decreased even when the content ratio of the PDA was decreased. As a result, it was confirmed that the time for coating with a thickness of 1 to 30 nm was 4 to 36 hours.

In order to coat the nanoparticles with a polymer material binding to a thiol group, silica shell nanoparticles (NPs@SiO ₂ ) having a thiol group were used, and as the polymer material, polyethylene glycol, polyetherimide, polyvinyl alcohol, casein , dextran, and chitosan were used. Silica shell nanoparticles were obtained by adding 0.5 mL of NH ₄ OH solution containing Tetra Ethyl Ortho Silicate (TEOS) to 80 mg of magnetic nanoparticles (MNP) or polystyrene nanoparticles such as iron oxide, and reacting for 16 hours to obtain silica. Nanoparticles with shells were prepared. Then, 40 mL of mercaptoethanol (MA, 80% v/v), 400 mg of EDC (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide), 40 mL of dimethylformamide ( DMF) and 80 mg 4-dimethylaminopyridine (DMAP) were added and reacted for 16 hours to form a thiol group on the silica shell surface. After that, 2 mg of silica shell nanoparticles having a thiol group on the surface and 0.6 mg of PEG powder (molecular weight: 6,000), PEI powder, dextaran, casein, or chitosan are added to 1 mL of phosphate buffer solution (pH 7.4) and mixed. After that, the reaction was carried out for 16 hours to prepare nanoparticles coated with a polymer material. At this time, the content ratio of the nanoparticles and the polymer material was variously changed, and nanoparticles coated with a polymer material were prepared. Nanoparticles were prepared, and when the weight ratio was greater than 1:5, the nanoparticles were agglomerated with each other, and when the weight ratio was less than 1:1, it was confirmed that the coating of the polymer material was not sufficient. And it was confirmed that the coating thickness of the polymer material was decreased as the reaction time was reduced to 12 hours, 8 hours, and 4 hours, and the coating thickness of the polymer material was decreased even when the content ratio of the polymer material was decreased.

Then, 2 mg of EDC (1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide) and 2 mg of NHS (N-hydroxysuccinimide) were added to the prepared polymer-coated nanoparticles so that the receptor could bind, and 6 A linker for amine coupling was bound by reaction at room temperature for a period of time. Then, after collecting the particles through a permanent magnet, the remaining solution is removed, the collected particles are added to 40 mL of phosphate buffer solution (pH 7.4), vortexed for 1 minute, and again, only the particles are collected using a permanent magnet. Washed 3 times. And finally, the nanoparticles coated with the polymer material prepared in 1 mL of phosphate buffer were stored and stored.

Hereinafter, uncoated silica nanoparticles were used as a control in all experiments. Nanoparticles coated with each polymer material prepared by the above method were observed using a scanning electron microscope (SEM). The results are shown in FIG. 1 .

As shown in FIG. 1 , in the case of the uncoated nanoparticles and the nanoparticles coated with a polymer material, it was confirmed that the surface feel was different, and through this, it was confirmed that the nanoparticles were coated with a polymer material. And among the nanoparticles coated with a polymer material, in the case of the nanoparticles coated with casein or dextran, it was confirmed that the nanoparticles were agglomerated, and in particular, it was confirmed that the agglomeration phenomenon was the largest in the case of casein. On the other hand, in the case of PDA, PEG, PEI, and chitosan, it was confirmed that there was almost no agglomeration phenomenon.

In addition, in order to observe the nanoparticles coated with the polymer material in more detail, the nanoparticles coated with PDA or PEG were observed using a transmission electron microscope (TEM). The results are shown in FIG. 2 .

As shown in FIG. 2 , it was confirmed that a thin film was formed on the surface of the nanoparticles coated with PDA or PEG, and it was confirmed that no film was formed on the silica nanoparticles as a control. Through the above results, the nanoparticles were coated with PDA or PEG to form a film, and through this, it was confirmed that PDA-coated nanoparticles (NPs@PDA) and PEG-coated nanoparticles (MNPs@SiO2-PEG) were normally prepared.

Example 2: Preparation of Nanoparticle Composites

In order to separate, purify and detect a target from a biological sample, pathogens, cells, biomarkers (nucleic acids, enzymes, proteins, etc.) Nanoparticle complexes were prepared in which a receptor capable of being bound to each other was obtained.

After adding the antibody or antibiotic to the PDA-coated nanoparticles, the PDA-coated nanoparticle complex was prepared by dispersing at 4° C. and reacting for 2 hours or more, in which the antibody or antibiotic was bound to the PDA. A method for preparing the PDA-coated nanoparticle complex to which an antibody or antibiotic is bound is briefly shown in FIG. 3 .

In order to bind a lectin that targets a virus, mannose (Man), a type of monosaccharide, and PDA are mixed in a weight ratio of 1:1, then iron oxide nanoparticles are added and sonicated for 16 hours. Man and PDA-coated nanoparticles (NPs@PDA-Man) were prepared through self-polymerization while dispersing, and Concanavalin A (ConA) was added to the prepared coated nanoparticles and reacted for 3 hours. To prepare a nanoparticle complex coated with Man and PDA covalently bound to lectin. The manufacturing method of the lectin-coupled Man/PDA-coated nanoparticle complex is briefly shown in FIG. 4 .

In order to bind antibiotics or antibodies to the PEG-coated nanoparticles to which the EDC/NHS linker is bonded, antibiotics such as vancomycin, polymyxin B, etc. are treated, and reacted at 4° C. for 2 hours. A nanoparticle composite was prepared. The method for preparing the PEG-coated nanoparticle complex to which an antibody or antibiotic is bound is briefly shown in FIG. 5 .

The prepared nanoparticle complexes were each stored in a phosphate buffer solution until use.

Example 3: Characterization of nanoparticle composites

3.1. Confirmation of dispersibility of nanoparticles

In order to confirm the dispersibility of nanoparticles prepared in the same manner as in Example 1 in a biological sample, PEG-coated silica nanoparticles (NPs@SiO ₂ -PEG) and PDA-coated nanoparticles that were confirmed to have the least agglomeration phenomenon (NPs@PDA) was added to whole blood, respectively, and the degree of aggregation between particles was checked according to the number of washings. More specifically, each nanoparticle is added to 1 mL of phosphate buffer solution, vortexed for 15 seconds, and then the nanoparticles are collected using a magnetic rack, then the supernatant is removed and the phosphate buffer solution is added again. The process was repeated 3 times. As a control, nanoparticles that were not coated were used. The results are shown in FIG. 6 .

As shown in FIG. 6 , it was confirmed that uncoated silica nanoparticles were aggregated and settled to the bottom, whereas nanoparticles coated with PEG or PDA were dispersed in blood and did not aggregate. Through the above results, it was confirmed that the nanoparticles coated with the polymer material of the present invention did not have agglomeration between the nanoparticles, as well as the matrix effect in the biological sample, as observed in the SEM.

3.2. Check target resolution

In order to confirm the target resolving ability of the nanoparticle complexes prepared in the same manner as in Example 2, the resolving ability of bacteria contained in whole blood was checked. In more detail, each pathogen or virus was mixed with whole blood, and the nanoparticle complexes were treated and reacted for 20 minutes, then the nanoparticle complexes were separated. And the amount of pathogens bound to the separated nanoparticle complex is confirmed by PCR using a bacterial universal 16s rDNA primer, and based on the amount of pathogens in the sample to which nanoparticles are not added, The amount of pathogen was expressed in %. The target strain is Escherichia coli O157:H7; E. coli), Staphylococcus aureus (S. aureus), antibiotic-resistant strain methicillin-resistant Staphylococcus aureus (MRSA), Kleb Klebsiella pneumonia (K. pneumonia), Pseudomonas aeruginosa (P. aeruginosa) and Salmonella enteritidis (S. enteritidis) were used, and nanoparticles were applied to uncoated silica nanoparticles. Receptor-coupled nanoparticle complex, PDA-coated nanoparticles with receptor bound nanoparticle complex, EDC/NHS linker bound PEG-coated nanoparticles with receptor bound nanoparticle complex, dextran-coated nanoparticles with receptors A nanoparticle complex bound to the nanoparticles and a nanoparticle complex in which a receptor was bound to PEI-coated nanoparticles were used. As the receptor, vancomycin or polymyxins B was used. The results are shown in FIG. 7 .

As shown in FIG. 7 , in the case of nanoparticles not coated with a polymer material, a capturing efficiency of 40% or less was exhibited, whereas in the case of nanoparticles coated with a polymer material, a capturing efficiency of 40% or more was obtained. was confirmed to be indicated. And among them, in the case of nanoparticles coated with PDA or PEG, it was confirmed that the capturing efficiency was 80% or more within 20 minutes, showing the highest capturing efficiency.

In addition, in order to confirm the resolution using the antibody, the influenza virus, Influenza A virus subtype H1N1, was mixed in saliva, and the PDA-coated nanoparticle complex to which the Anti-H1N1 Antibody (Santa Cruz Biotechnology) was bound was treated and reacted for 20 minutes. Then, the nanoparticle complex was separated. And the amount of virus bound to the separated nanoparticle complex was confirmed using the antibody. As a result, when the virus is isolated using the PDA-coated nanoparticle complex to which the Anti-H1N1 antibody is bound, about 90% of the virus is isolated within 20 minutes, whereas the non-coated nanoparticle complex to which the Anti-H1N1 antibody is bound. In the case of using , about 40% of the virus was isolated, and it was confirmed that the collection efficiency was significantly increased in the case of nanoparticles coated with a polymer material.

Through the above results, the nanoparticles coated with the polymer of the present invention not only inhibit the aggregation between nanoparticles in aqueous solution, but also prevent the matrix effect in biological samples in which various components such as blood, saliva, and urine are present in a complex manner. Since the particles can exist in an efficiently dispersed state by inhibiting aggregation between particles, it is possible to significantly increase the separation and purification efficiency of the target from a practical biological sample, and through this, the detection sensitivity, detection accuracy, and diagnostic accuracy of the analysis technology finally It was confirmed that the back can be significantly increased. In addition, it was confirmed that various types of receptors can be easily bound without limitation to the surface of the coated polymer material through various known methods, so that it can be effectively applied to various fields of detecting various target substances.

The foregoing description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (23)

A nanoparticle complex for detecting or separating a target material from a biological sample, comprising:
In the nanoparticle composite, the surface of the nanoparticles is coated with a polymer material (macromolecule),
A receptor for detecting or separating a target material is bound to the polymer material,
The polymer material is made of polydopamine, polyethylene glycol, polyetherimide, polyvinyl alcohol, casein, dextran, and chitosan. Nanoparticle composite, characterized in that at least one selected from the group. The method of claim 1,
The coating is a nanoparticle complex, characterized in that a monosaccharide for lectin binding is mixed with a polymer material and coated. 3. The method of claim 2,
The monosaccharide is mannose (mannose), glucose (glucose), fructose (fructose), and glucosamine (glucosamine), characterized in that any one or more selected from the group consisting of, the nanoparticle complex. The method of claim 1,
The bond is an ionic bond, a covalent bond, a coordination bond, a hydrogen bond, a hydrophobic interaction, a van der Waals interaction, and any one or more types selected from the group consisting of a linker, the nanoparticle complex. 5. The method of claim 4,
The binding by the linker is 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS), characterized in that the binding by the linker or a thiol group, the nanoparticle complex. The method of claim 1,
The receptor is a nanoparticle complex, characterized in that any one or more selected from the group consisting of antibiotics, antibodies, nucleic acids, lectins, and proteins that specifically bind to a target substance. The method of claim 1,
The target material is a pathogen, virus, cell, protein, nucleic acid, antigen, characterized in that any one or more selected from the group consisting of a glycoprotein, nanoparticle complex. The method of claim 1,
The nanoparticles are silica nanoparticles, iron oxide nanoparticles, and any one or more nanoparticles selected from the group consisting of polystyrene nanoparticles, the nanoparticle composite. The method of claim 1,
The nanoparticle composite has a diameter of 200 to 1,000 nm, characterized in that the nanoparticle composite. The method of claim 1,
The biological sample is blood, saliva, bone marrow fluid, lymph fluid, urine, amniotic fluid, mucosal fluid, and peritoneal fluid, characterized in that any one or more selected from the group consisting of, the nanoparticle complex. A method for preparing the nanoparticle composite of claim 1, characterized in that it comprises the following steps:
a) preparing polydopamine-coated nanoparticles by adding and dispersing nanoparticles and an acid-based catalyst to a polydopamine solution; and
b) preparing a polydopamine-coated nanoparticle complex to which a receptor is bound by adding and reacting a receptor to the polydopamine-coated nanoparticles. 12. The method of claim 11,
The nano-particles and polydopamine of step a) are 1: 0.01 to 1: 1.5, characterized in that mixed in a weight ratio, the manufacturing method. 12. The method of claim 11,
A manufacturing method, characterized in that the monosaccharide is further added to the polydopamine solution of step a) in a weight ratio of 1: 0.5 to 1: 1.5. 12. The method of claim 11,
The nanoparticles are iron oxide nanoparticles or polystyrene nanoparticles, characterized in that the manufacturing method. 12. The method of claim 11,
Step a) is characterized in that the dispersion for 4 hours to 36 hours, the manufacturing method. 12. The method of claim 11,
Step b) is characterized in that the reaction for 1 hour to 5 hours, the production method. A method for preparing the nanoparticle composite of claim 1, characterized in that it comprises the following steps:
a) preparing nanoparticles coated with a polymer material by adding and dispersing a polymer material to silica shell nanoparticles having a thiol group; and
b) adding and reacting a receptor to the coated nanoparticles to prepare a nanoparticle complex coated with a polymer material to which the receptor is bound,
The polymer material is any one selected from the group consisting of polyethylene glycol, polyetherimide, polyvinyl alcohol, casein, dextran, and chitosan. The above, characterized in that the manufacturing method. 18. The method of claim 17,
In step b), EDC (1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide) and NHS (N-hydroxysuccinimide) are added together with the receptor, the manufacturing method. 18. The method of claim 17,
The nano-particles and the polymer material of step a) are 1:1 to 1:5, characterized in that mixed in a weight ratio, the manufacturing method. 18. The method of claim 17,
Step a) is characterized in that the mixing and dispersing for 4 hours to 36 hours, the manufacturing method. 18. The method of claim 17,
Step b) is characterized in that the reaction for 1 hour to 5 hours, the manufacturing method. A kit for detecting or separating a target material from a biological sample, comprising the nanoparticle complex of claim 1 as an active ingredient. A kit for diagnosing infection, comprising the nanoparticle complex of claim 1 as an active ingredient.

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