TWI665441B - Janus particle, tetrahedral structure including janus particles, method of fabricating janus particles, and method of detecting biomolecules - Google Patents

Abstract

A bilateral asymmetric particle (Janus particle) comprising a low-dimensional substrate and a plurality of biomolecules, the surface of the low-dimensional substrate comprising a biomolecule modified region, the biomolecule being fixed to the surface of the low-dimensional substrate and located at the biomolecule In the modified region, the area between the biomolecule modified region and the area of the low-dimensional substrate has the following relationship: (1/5) AS≦AB≦(1/2)AS

Among them, AB represents the total area of the biomolecule modified region, and AS represents the total area of the low-dimensional substrate.

Description

 Bilateral asymmetric particles, tetrahedral polymeric structure with bilateral asymmetric particles, method for producing bilateral asymmetric particles, and method for detecting biomolecule  

The invention relates to a structure and a manufacturing method of bilateral asymmetric particles, in particular to a structure and a manufacturing method of bilateral asymmetric particles for detecting microorganisms.

Recently, due to the increasing frequency of cross-domain activities of human beings, the speed of infection and the rate of mutation of pathogenic sources of animals and plants have also increased. In the Asia-Pacific region, for example, adenovirus, influenza virus, Zika virus, dengue fever and other regional infectious diseases. It often erupts in certain seasons, so it is increasingly important to construct sensitive, rapid, quantifiable, and low-cost detection and diagnostic methods.

At present, the commonly used detection methods mainly include immunological detection methods and methods such as molecular biology. For immunological detection methods, it mainly includes an enzyme-linked immunosorbent assay (ELISA) and a gold immunochromatography assay (GICA). Among them, the enzyme immunoassay has the advantages of being able to be roughly quantitatively tested, simple in method and high in sensitivity, and thus can be used to determine a large number of trace samples. However, since it involves multiple washing steps, the steps are cumbersome and time consuming. Colloidal gold immunochromatography has the advantages of rapid measurement, simple method and low cost, but its sensitivity is relatively low, so it is still difficult to quantify the sample to be tested. Regarding the detection method of molecular biology, it mainly uses a polymerase chain reaction (PCR) to perform microbial detection, which has the advantages of high detection sensitivity and good specificity. However, the implementation of this test method must use expensive equipment and skilled technicians, and because the biochemical reaction requires multiple cycles, it is also time consuming to use. In addition, the current virus screening time requires at least about 30 minutes, and the accuracy is only about 6 to 70%. The screening results are often only available to medical professionals as a reference, and medical professionals still need to use other test results. Comprehensively determine whether it is infected by a virus. In summary, the accuracy and timeliness of conventional techniques for testing are still not perfect, so it is still necessary to develop a detection technique to solve the above-mentioned shortcomings in the prior art.

According to an embodiment of the present invention, there is provided a bilateral asymmetric particle comprising a low-dimensional substrate and a plurality of biomolecules, the surface of the low-dimensional substrate comprising a biomolecule modified region, and the biomolecule is fixed to the low-dimensional basis The surface of the material is located in the biomolecule modification region, wherein the area of the biomolecule modified region and the area of the low dimensional substrate have the following relationship: (1/5) AS≦AB≦(1/2)AS. Among them, AB represents the total area of the biomolecule modified region, and AS represents the total area of the low-dimensional substrate.

According to another embodiment of the present invention, there is provided a tetrahedral polymer structure comprising a microorganism and four bilateral asymmetric particles surrounding the microorganism, the surface of the microorganism comprising a plurality of biomolecules, and each bilateral asymmetric particle Including a low-dimensional substrate and a plurality of other biomolecules, the surface of the low-dimensional substrate includes a biomolecule modification region, and each of the other biomolecules is located in the biomolecule modification region, and the two biomolecules are Each biomolecule is fixed to the surface of the low-dimensional substrate and the surface of the microorganism.

According to another embodiment of the present invention, there is provided a method of fabricating a bilateral asymmetric particle comprising the steps of: providing at least one low dimensional substrate, adsorbing a low dimensional substrate on a surface of the fibrous network; performing heating The process causes the low-dimensional substrate portion to sag into the fibrous network structure to form a depressed portion, and the portion of the low-dimensional substrate protrudes from the surface of the fibrous network structure to form a protruding portion, wherein the protruding portion The ratio of the area to the area of the low-dimensional substrate is between 0.2 and 0.5; forming a surface modifying layer on the surface of the protrusion; after forming the surface modifying layer, the low-dimensional substrate is formed on the surface of the fiber network Disengaging; and providing a biomolecule layer on the surface modifying layer, wherein the biomolecule layer is fixed to the surface modifying layer.

According to still another embodiment of the present invention, there is provided a method of detecting biomolecules comprising the steps of: first, providing a plurality of bilateral asymmetric particles, each bilateral asymmetric particle comprising a low dimensional substrate and a plurality of biomolecules. The surface of the low-dimensional substrate comprises a biomolecule modified region, and the biomolecules each comprise a fixed end and a free end, each fixed end being fixed to the surface of the low-dimensional substrate and located in the biomolecule modified region, wherein The area between the biomolecule-modified region and the area of the low-dimensional substrate has the following relationship: (1/5) AS≦AB≦(1/2)AS. Wherein, AB represents the total area of the modified region of the biomolecule, and AS represents the total area of the low-dimensional substrate; then, a microorganism is provided, which includes a plurality of other biomolecules, which are disposed on the surface of the microorganism, and finally, bilaterally The asymmetric particles and the microorganisms are mixed with each other such that the free ends of the respective biomolecules located on the surface of the bilateral asymmetric particles are fixed to the respective biomolecules on the surface of the microorganism.

According to an embodiment of the invention, the low-dimensional substrate is a spherical substrate having a mesopores.

According to an embodiment of the invention, the biomolecule modification region is a continuous region and is disposed on one side of the low dimensional substrate.

According to an embodiment of the invention, the biomolecule is uniformly distributed in the biomolecule modification region.

According to an embodiment of the invention, each of the biomolecules described above is one of an antibody or an antigen.

According to an embodiment of the invention, the surface of the low-dimensional substrate further includes a non-biomolecular modification region, and the non-biomolecular modification region is a continuous region and is disposed on one side of the low-dimensional substrate.

According to an embodiment of the present invention, the area of the non-biomolecule modified region is represented by the following formula: AM = AS-AB. Where AM represents the total area of the non-biomolecule modified region.

According to an embodiment of the invention, the low-micron substrate further includes a plurality of holes, and the fluorescent substance is disposed in the holes.

According to an embodiment of the invention, the bilateral asymmetric particles further comprise a plurality of fluorescent molecules fixed to the surface of the low-dimensional substrate and located in the biomolecule modification region.

According to an embodiment of the invention, the fluorescent molecule is a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor, or a single fluorescent molecule.

According to an embodiment of the present invention, each of the bilateral asymmetric particles further includes a first fluorescent molecule, each of the first fluorescent molecules is fixed to the biomolecule modified region of the surface of the low-dimensional substrate, and the microorganism further includes the second Fluorescent molecules, each of the second fluorescent molecules is immobilized on the surface of the microorganism, and each of the first fluorescent molecules is one of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor, and each of the second The fluorescent molecule is the other of the fluorescent resonance energy transfer donor or the fluorescent resonance energy transfer acceptor.

According to an embodiment of the present invention, a part of the particles in the bilateral asymmetric particles further includes a first fluorescent molecule, and each of the first fluorescent molecules is fixed on the surface of the low-dimensional substrate and located in the modified region of the biomolecule, bilaterally The other particles in the asymmetric particle further comprise a second fluorescent molecule, each second fluorescent molecule is fixed on the surface of the low-dimensional substrate and located in the modified region of the biomolecule, and each of the first fluorescent molecules is a fluorescent resonance One of the energy transfer donor or the fluorescent resonance energy transfer acceptor, each of the second fluorescent molecules being the other of the fluorescent resonance energy transfer donor or the fluorescent resonance energy transfer acceptor.

According to an embodiment of the invention, the bilateral asymmetric particles further comprise a magnetic substance disposed in the non-bio-modified region.

According to an embodiment of the invention, the magnetic substance is disposed in the hole.

According to an embodiment of the present invention, each of the bilateral asymmetric particles in which the surface is fixed with the first fluorescent molecules further comprises a magnetic substance disposed in the non-bio-modified region of the low-dimensional substrate, and the surface is fixed with the second fluorescent material. Each bilateral asymmetric particle of the optical molecule does not include any magnetic species.

According to an embodiment of the present invention, the number of bilateral asymmetric particles in which the surface is fixed with the first fluorescent molecules is the same as the number of bilateral asymmetric particles in which the second fluorescent molecules are fixed on the surface.

According to an embodiment of the present invention, each biomolecule on the surface of the low-dimensional substrate has a specific binding force to each biomolecule located on the surface of the microorganism.

According to an embodiment of the invention, the fibrous network structure is composed of at least one electrospun fiber, and the segments of the electrospun fiber are stacked on each other.

According to an embodiment of the present invention, the method for fabricating the bilateral asymmetric particles further includes the steps of: forming a protective layer on the surface modifying layer before the low-dimensional substrate is detached from the surface of the fibrous network structure; After the dimensional substrate is detached from the surface of the fibrous network structure, the protective layer is removed.

According to an embodiment of the present invention, the diameter of the low-dimensional substrate and the diameter of the microorganism have the following relationship: 0.15 ≦ (VD/D) ≦ 0.3. Wherein D represents the diameter of the low dimensional substrate and VD represents the diameter of the microorganism.

100‧‧‧ bilateral asymmetric particles

102‧‧‧Low dimension substrate

104‧‧‧Biomolecules

106‧‧‧ Magnetic materials

108‧‧‧Fluorescent molecules

120‧‧‧Sag

122‧‧‧Protruding

130‧‧‧Biomolecular modification area

132‧‧‧Non-biomolecular modification areas

200‧‧‧Microorganisms

202‧‧‧Biomolecules

300‧‧‧tetrahedral aggregate structure

400‧‧‧Fiber mesh structure

402‧‧‧Electrastic fiber

Figure 1 is a schematic illustration of bilateral asymmetric particles in accordance with one embodiment of the present invention.

Fig. 2 is a tetrahedral polymerization structure according to an embodiment of the present invention.

Figure 3 is a schematic illustration of a low dimensional substrate adsorbed onto the surface of a fibrous network structure in accordance with one embodiment of the present invention.

Fig. 4 is a schematic view showing the addition of a test object to a solution containing bilateral asymmetric particles according to an embodiment of the present invention.

Figure 5 is an electron micrograph of a self-assembled tetrahedral polymer structure prepared in accordance with one embodiment of the present invention.

Fig. 6 is a particle size distribution diagram of an embodiment of the present invention.

Fig. 7 is a particle size distribution diagram of an embodiment of the present invention.

Figure 8 is a graph showing the fluorescence resonance energy transfer intensity distribution of different simulated virus concentrations in an embodiment of the present invention.

Fig. 9 is a view showing the effect of a tetrahedral polymeric structure magnetized by a magnet on the fluorescence resonance energy transfer intensity according to an embodiment of the present invention.

In the following, specific embodiments of bilateral asymmetric particles, tetrahedral polymeric structures with bilateral asymmetric particles, methods for making bilateral asymmetric particles, and methods for detecting biomolecules are presented, which have the general technology in the art. The invention can be implemented accordingly. The specific embodiments may be referred to the corresponding drawings, such that the drawings form part of the embodiments. Although the embodiments of the present invention are disclosed as follows, they are not intended to limit the invention, and those skilled in the art can make some modifications and refinements without departing from the spirit and scope of the invention. Here, the methods used in the respective examples and experimental examples are conventional methods unless otherwise specified. In addition, the materials and reagents used in the respective examples and experimental examples were mainly purchased from Sigma-Aldrich, and other materials and reagents were commercially available from conventional biochemical reagent suppliers unless otherwise specified.

Figure 1 is a schematic illustration of bilateral asymmetric particles in accordance with one embodiment of the present invention. Referring to FIG. 1, a bilateral asymmetric particle 100 comprises a low dimensional substrate 102 and a plurality of biomolecules 104. The surface of the low-micron substrate 102 includes a biomolecule modification region 130, and the biomolecule 104 is fixed to the surface of the low-dimensional substrate 102 and located in the biomolecule modification region 130, wherein the area and low dimension of the biomolecule modification region 130 The area of the substrate 102 has the following relationship: (1/5) AS≦AB≦(1/2)AS, preferably, (1/4) AS≦AB≦(1/3)AS. Among them, AB represents the total area of the biomolecule modified region, and AS represents the total area of the low-dimensional substrate.

It should be noted that the term "bilateral asymmetric particles" as used throughout refers to a region of the particle surface having at least two mutually different chemical properties and/or physical properties. For example, one side of the bilateral asymmetric particles 100 may The specific antigen is specific, magnetic, fluorescent, and/or has fluorescence resonance energy transfer (FRET), while the other side of the bilateral asymmetric particle 100 is not specific to the antigen. It is specific, non-magnetic and/or non-fluorescent.

The low dimensional substrate 102 described above means that the dimensions of the substrate in each axial direction are less than 1000 nm, and preferably less than 500 nm. The low-dimensional substrate 102 may be a spherical substrate, a columnar substrate, or a dumbbell-shaped substrate, but is not limited thereto, and its composition may be selected from the group consisting of ceramic materials or biocompatible molecules. According to an embodiment, the low dimensional substrate 102 can be a ceria spherical substrate having a diameter between 10 and 1000 nanometers, and preferably 500 nanometers. In addition, depending on the use requirements, the surface layer and the interior of the low-dimensional substrate 102 may also have pores. For example, it may be a mesoporous spherical ceria substrate having a pore diameter of 2 -50 nm between.

The biomolecule 104 is selected from the group consisting of a protein, a peptide, an amino acid, a nucleic acid or other suitable biomolecule. Preferably, the biomolecule 104 is avidin or has specific binding to a specific antigen. Antibody. One end of the biomolecule 104 is fixed to the surface of the low-dimensional substrate 102 by chemical bonding. In the case where the biomolecule 104 is selected from avidin, the surface of the low-dimensional substrate 102 preferably has an amine group (-NH ₂ ) functional group, such that the amine group and the carboxyl group (-COOH) in the biotin. An amide bond is produced. In addition, according to other embodiments, an epoxy epoxide may be attached to the surface of the low-dimensional substrate 102, and an epoxy functional group may be reacted with an amino group (-NH ₂ ) in the biomolecule to generate carbon- Nitrogen (CN) bonding. It should be noted that the above bonding method is merely an example, and the bonding method should not be limited to the above manner.

The biomolecule modification region 130 described above is a continuous region distributed on one side of the low-dimensional substrate 102. In other words, the biomolecular modification region 130 does not occupy the entire surface of the low dimensional substrate 102. In the case where the biomolecule 130 is only disposed in the biomolecule modification region 130, the surface of the low-dimensional substrate in which the biomolecule 130 is not provided may also be referred to as a "non-biomolecular modification region".

According to an embodiment of the invention, the surface of the bilateral asymmetric particles 100 may further comprise a non-biomolecular modification region 132, which may be a continuous region and located on different sides of the bilateral asymmetric particles 100. For example, the non-biomolecule modified region 132 and the biomolecule modified region 130 may be located on different sides of the bilateral asymmetric particles 100, respectively, such that the two regions do not overlap each other. Preferably, the boundary between the non-biomolecular modification region 132 and the biomolecular modification region 130 is flush with each other, in which case the low dimensional substrate (AS), the non-biomolecular modification region (AM), and the organism The area relationship between the molecular modification regions (AB) is: AM = AS-AB. Further, a specific material such as the magnetic material 106 may be provided in the non-biomolecule modification region 132, but is not limited thereto. In particular, the magnetic material 106 is preferably a material having a magnetic dipole, such as a paramagnetic or ferromagnetic material, such that the magnetic material 106 can exhibit a particular magnetic properties in a magnetic field. In addition, for the bilateral asymmetric particles 100 having holes, the magnetic material 106 is disposed outside the surface of the bilateral asymmetric particles 100, and is also disposed in each of the holes of the bilateral asymmetric particles 100, and The system is placed on the side wall of each hole.

According to an embodiment of the invention, the bilateral asymmetric particles 100 may further comprise fluorescent molecules 108 fixed to the surface of the low-dimensional substrate 102 and located in the biomolecule modification region 130. The fluorescent molecule 108 can be a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor or a single fluorescent molecule.

Fig. 2 is a tetrahedral polymerization structure according to an embodiment of the present invention. According to an embodiment of the invention, the bilateral asymmetric particles 100 may form a tetrahedral polymeric structure 300 with the microorganisms 200, each tetrahedral polymeric structure 300 comprising a single microbial organism 200 and four bilateral asymmetric particles surrounding the microbial organism 200. 100. The microorganisms 200 are located in a tetrahedral hole surrounded by four bilateral asymmetric particles 100, and the bilateral asymmetric particles 100 each constitute the apex angle of the tetrahedral polymeric structure 300, preferably, bilaterally The symmetric particles 100 are equally spaced from each other, but are not limited thereto. The surface of the microorganism 200 includes a plurality of biomolecules 202, and one end of each biomolecule 104 on the surface of the bilateral asymmetric particle 100 is fixed to the surface of the low-dimensional substrate 102, and the other end is fixed to each organism on the surface of the microorganism 200. Molecule 202. Preferably, the relationship between the longest dimension of the low-dimensional substrate (D) along a single axial direction and the longest dimension of the microorganism (VD) along a single axial direction is: 0.1 ≦ (VD/D) ≦ 0.35, The best is between 0.15 ≦ (VD/D) ≦ 0.3. It should be noted that the term "microorganism" as used throughout the text refers to an organism having a genetic material and having self-replication ability, such as a virus, having a longest dimension along a single axial direction of 20-150 nm, but is not limited thereto. .

It should be noted that for the case where the bilateral asymmetric particles 100 include fluorescent molecules 108, the surface of the microorganisms 200 within the respective tetrahedral polymeric structures 300 will also have corresponding fluorescent molecules. For example, when the fluorescent molecules 108 located on the surface of the bilateral asymmetric particles 100 are selected from one of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor, the surface of the microorganism 200 is solidified. The other of the fluorescent resonance energy transfer donor or the fluorescent resonance energy transfer acceptor is connected.

Further, the above-mentioned bilateral asymmetric particles 100 may also constitute an octahedral polymerization structure with the microorganisms 200. Specifically, each octahedral polymeric structure is composed of a single microorganism 200 and six bilateral asymmetric particles 100 such that the microorganism 200 is located in a central pore formed by six bilateral asymmetric particles 100 (octahedral hole) )in.

According to the above embodiment, bilateral asymmetric particles 100 and a tetrahedral polymeric structure 300 comprising bilateral asymmetric particles 100 are disclosed. The following is a description of how to make bilateral asymmetric particles, and it can be used with reference to Figure 1.

According to an embodiment of the invention, a method of making bilateral asymmetric particles is provided. First, at least one low dimensional substrate is provided, and the substrate can be selected from the group consisting of ceramic substrates or biocompatible molecules. For example, the low dimensional substrate is a ceria sphere and has a particle size (D) between 10 and 1000 nm, and preferably 500 nm.

Thereafter, holes can be selectively formed in the low dimensional substrate. For example, for a spherical substrate composed of cerium oxide, a protective layer such as polyvinylpyrrolidone (PVP) may be formed on the surface of the spherical substrate, and then etched with an etching solution to cover the unprotected layer. The spherical substrate is formed, and a hole is formed in the spherical substrate, preferably a mesoporous.

Referring to Figure 3, there is shown a schematic view of a low dimensional substrate adsorbed onto the surface of a fibrous network structure in accordance with one embodiment of the present invention. Next, the low dimensional substrate 102 is adsorbed onto the surface of the fibrous web 400. The fiber mesh structure 400 may be formed by alternately stacking electrospun fibers 402. The composition of the electrospun fibers 402 may be a single polymer or a plurality of polymers, and each polymer may be a homopolymer or a copolymer ( Copolymer). For example, the composition of the electrospun fiber 402 may be selected from the group consisting of an acrylic polymer, a vinyl polymer, a polyester, and a polyamide, but is not limited thereto. . Preferably, the electrospun fiber 402 is composed of two kinds of polymers, and the composition thereof is polymethylmethacrylate (PMMA) and poly(4-vinylpyridine) (P4VP). ), but not limited to this.

Next, still referring to FIG. 3, a heating process is performed to cause the low-dimensional substrate 102 to partially sink into the fibrous network structure 400 (the area depressed in the fiber mesh structure 400 may be referred to as the lower trap 120). A portion of the low-dimensional substrate 102 is protruded from the surface of the fibrous web 400 (the region protruding from the fibrous web 400 may be referred to as a protrusion 122). The ratio of the area of the protruding portion 122 to the area of the low-dimensional substrate 102 is between 0.2 and 0.5. In other words, the area of the protruding portion 122 may be smaller than the area of the depressed portion 120.

Next, a deposition process, such as a chemical vapor deposition process or a liquid phase chemical reaction, is performed to form a surface modifying layer on the surface of the protrusion 122 of the low-dimensional substrate 102. At this point in the process, since the depressed portion 120 of the low-dimensional substrate is still shielded by the electrospun fibers 402, the surface modifying layer does not cover the depressed portion 120 of the low-dimensional substrate. According to an embodiment of the present invention, the surface modifying layer is composed of organic molecules, one end of which is bonded to the surface of the low-dimensional substrate 102, and the other end of which has a specific functional group such as an amine group. This amine group can be bonded to the layer of biomolecules 104 that are subsequently disposed on the surface of the low-dimensional substrate 102, causing the biomolecules 104 to be anchored to the low-dimensional substrate 102. The organic molecule in the surface modifying layer described above may be selected from 3-aminopropyltrimethoxysilane (APS), but is not limited thereto.

Thereafter, another deposition process, such as a vapor deposition process, is performed to form a protective layer covering the surface modifying layer, such as paraffin wax. At this point in the process, since the depressed portion 120 of the low-dimensional substrate 102 is still shielded by the electrospun fibers 402, the protective layer also does not cover the depressed portion 120 of the low-dimensional substrate 102.

Thereafter, the fibrous web 402 is removed using an organic solvent or other suitable method to cause the low dimensional substrate 102 to detach from the surface of the fibrous web 402. At the time of preparation, a surface modification layer and a protective layer are sequentially stacked on one side of each low-dimensional substrate 102, and the surface modification layer and the protective layer are not provided on the other side of each low-dimensional substrate. .

Next, a specific synthesis method, such as a sol-gel, can be selectively applied in the liquid phase to synthesize the desired magnetic material 106 in the surface of the low-dimensional substrate 102 and/or its pores. The magnetic material 106 may be selected from a ferromagnetic material or a ferromagnetic material, preferably a nanoparticle of iron oxide (Fe ₂ O ₃ or Fe ₃ O ₄ ). During this synthesis, since a particular region of the low-dimensional substrate 102 is still covered by the protective layer, the magnetic material 106 will only be synthesized in the region not covered by the protective layer. Thereafter, the low-dimensional substrate 102 is washed with a solvent such as hexane to remove paraffin remaining on the surface to expose the surface modifying layer originally under the paraffin.

Next, the biomolecule modification region 130 on the low-dimensional substrate 102 can be selectively attached to the fluorescent molecule 108, and the fluorescent molecule 108 can be selected from a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor. In one embodiment, according to an embodiment of the invention, the fluorescent molecule 108 is a fluorescent resonance energy transfer donor fluorescent dye, such as a Marina Blue dye.

Thereafter, the biomolecules 104 are fixed in the biomolecule modification region 130 of the low-dimensional substrate 102, and the types of the biomolecules 104 are as described in the above embodiments, and are not described herein again. Thus, an immunologically active submicron bilateral asymmetrically modified magnetic material is obtained.

It should be noted that the timing of fixing the fluorescent molecules 108 and the biomolecules 104 to the low-dimensional substrate 102 is not limited to the above sequence. According to other embodiments, the order may also be reversed from each other. According to the above embodiment, the disclosure is disclosed. The method of making bilateral asymmetric particles is described below by using bilateral asymmetric particles to detect biomolecules. It should be noted that the bilateral asymmetric particles used in the embodiment can adopt the bilateral asymmetric particles prepared in the above embodiments, and thus the detailed structure and composition thereof can be referred to the above embodiments, and will not be described again.

Similar to the bilateral asymmetric particles of the corresponding embodiment of FIG. 1, each of the bilateral asymmetric particles 100 employed in this embodiment also includes a low dimensional substrate 102 and a plurality of biomolecules 104. The surface of the low-dimensional substrate 102 includes a biomolecule modification region 130, and the biomolecules 104 each include a fixed end and a free end, and each fixed end is fixed to the surface of the low-dimensional substrate 102 and is located in the biomolecule modification. In the region 130, the area between the biomolecule modified region (AB) and the area of the low dimensional substrate (AS) has the following relationship: (1/5) AS≦AB≦(1/2)AS, preferably , (1/4) AS≦AB≦ (1/3) AS.

Fig. 4 is a schematic view showing the addition of a test object to a solution containing bilateral asymmetric particles according to an embodiment of the present invention. Next, the analyte, such as the microorganism 200, is added to the solution containing the bilateral asymmetric particles 100. Since the biomolecules 202 on the surface of the microorganisms 200 and the free ends of the biomolecules 104 on the surface of the bilateral asymmetric particles 100 produce a specific combination, when the microorganisms 200 and the bilateral asymmetric particles 100 are mixed with each other, they are bilaterally not The free ends of the respective biomolecules 104 on the surface of the symmetric particles 100 can be attached to the respective biomolecules 202 on the surface of the microorganisms 200. In addition, when the fluorescent molecule 108 is modified on the surface of the bilateral asymmetric particle 100, such as one of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer receptor, the surface of the microorganism 200 is also modified. There are fluorescent molecules, such as the other one of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer receptor.

It should be noted that there is a specific relationship between the area of the biomolecule modified region (AB) and the area of the low dimensional substrate (AS): (1/5) AS≦AB≦(1/2)AS, and low dimension The longest dimension of the substrate along a single axis (D) has a specific relationship with the longest dimension (VD) of the microorganism along a single axis: 0.15 ≦ (VD/D) ≦ 0.3, so for a bilateral with a specific particle size In the case of the symmetrical particles 100, it can be combined with the microorganisms 200 having a specific particle diameter, and constitute a tetrahedral polymerization structure having a structure similar to that shown in the corresponding embodiment of Fig. 2. Specifically, every four bilateral asymmetric particles can capture a single microorganism having a specific particle size. Therefore, by analyzing the presence or absence and quantity of the tetrahedral polymerization structure by a specific instrument such as dynamic light scattering (DLS), the presence or absence and quantity of the above specific microorganisms can be determined.

Further, in order to allow each of the microorganisms 200 to be captured by the bilateral asymmetric particles 100, it is preferred to slowly add the microorganisms 200 to the solution containing the bilateral asymmetric particles 100, and the amount between the bilateral asymmetric particles 100 and the microorganisms 200. The ratio should be much larger than 10:1, more preferably 30:1 to 80:1.

It should be noted that the relationship between the area of the biomolecule modified region (AB) and the area of the low dimensional substrate (AS) is (1/2) AS<AB, because the area of the biomolecule modified region is larger. Large, when bilateral asymmetric particles and microorganisms are mixed with each other, the biomolecule modification region of a single bilateral asymmetric particle may combine with more than two microorganisms at the same time, and thus the tetrahedral stereostructure cannot be smoothly formed. In other words, it is impossible to capture microorganisms having a specific particle diameter by bilateral asymmetric particles, and it is also impossible to determine the number of microorganisms by the number of stable tetrahedral stereostructures.

In addition, since the bilateral asymmetric particles described above may selectively have magnetic materials and/or fluorescent molecules, and the surface of the microorganisms may be selectively modified with fluorescent molecules, in the detection process, The intensity of the signal generated by the optical resonance energy transfer is used to determine the number of tetrahedral polymeric structures, and the microorganisms are quickly separated by centrifugation or precipitation steps, and the presence or absence and quantity of the microorganisms are discriminated. In addition, when measuring the signal intensity of the fluorescence resonance energy transfer, a magnetic field may be additionally applied to the solution containing the tetrahedral polymer structure to aggregate the magnetic tetrahedral polymer structure, thereby increasing the signal intensity of the fluorescence resonance energy transfer.

In order to enable those of ordinary skill in the art to practice the invention, various preparations, specific examples and detection examples of the invention are described in further detail below. It is to be noted that the following specific examples are merely illustrative and the invention should not be construed as limiting. That is, the materials and materials used in the respective embodiments, the processing flow, and the like can be appropriately changed without departing from the scope of the invention.

Method for preparing bilateral asymmetric modified particles

Preparation Example 1 - Preparation method of bilateral asymmetric particles having immunological activity and fluorescent molecules

The five-nano-nano-cerium dioxide sphere is used as the main body, and it is washed with polyvinylpyrrolidone (PVP) polymer to protect the surface of the sphere, and is etched with sodium hydroxide to produce a medium-hole cerium oxide submicron sphere. Its particle size is maintained at 500 nanometers. Thereafter, the mesoporous silica sphere is adsorbed on the electrospinning structure, wherein the electrospinning composition in the electrospinning structure is: polymethylmethacrylate (PMMA) and poly(4-vinylpyridine). (Poly (4-vinylpyridine), P4VP). Then adjust the temperature to 158 degrees Celsius to immerse the mesoporous cerium oxide sphere in electrospinning, so that two-thirds of the sphere surface will be coated by electrospinning, and one-third of the sphere surface will be exposed. Electrospinning. Specifically, since the surface energy of the polymer fiber is automatically balanced with the sphere under a constant temperature state, the sphere is stopped at a certain amount of depression. Further, the surface energy of the above polymer fiber can be finely adjusted by adjusting the composition of the polymer material (for example, the ratio of PMMA and P4VP). By adjusting the surface energy and the temperature, it is possible to control the amount of depression in the polymer fiber to which the sphere is sunk. Then, a chemical vapor deposition process is performed to deposit 3-aminopropyltrimethoxysilane (APS) on the exposed sphere surface to modify the surface of the exposed sphere. A submicron bilateral asymmetric sphere (Janus particle) was produced. The electrospinning is subsequently dissolved in an organic solvent. The Marina Blue fluorescent dye is then attached to the surface of the one-third amine-modified sphere. Assisted IgG-type avidin antibody with N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) Anti-Biotin antibody) is affixed to one-third of the surface of the sphere to obtain immunologically active bilateral asymmetric particles, abbreviated as A-1.

Preparation Example 2 - Preparation method of bilateral asymmetric particles having immunological activity and fluorescent molecules

The preparation method of the preparation example is similar to the preparation method of the above preparation example 1, and the main difference is that 1/2 of the bilateral asymmetric particles in the preparation example have a fluorescence resonance energy transfer donor, such as Marina Blue, and the remaining 1 The /2 number of bilateral asymmetric particles will have a fluorescent resonance energy transfer acceptor such as 6-(7-Nitrobenzofurazan-4-ylamino)hexanoic acid. The bilateral asymmetric particles prepared in this preparation example are referred to as A-2.

Preparation Example 3 - Preparation method of bilateral asymmetric particles having immunologically active, magnetic and fluorescent molecules

The preparation method of the preparation example is similar to the preparation method of the above preparation example 1, and the main difference is that after the sub-micron bilateral asymmetric sphere is manufactured and before the Marina blue fluorescent dye is attached to the surface of the amine-modified sphere, The step of performing synthetic iron oxide. The detailed steps are as follows: After the submicron bilateral asymmetric spheres are fabricated, another vapor deposition process is performed to coat the exposed sphere surfaces with ultrapar wax. Subsequent electrolysis of the electrospinning with an organic solvent yields one-third of the bilateral asymmetric spherical spheres in which the surface of the sphere is coated with wax. Thereafter, the bilateral asymmetric spherical spheres of the mesopores are placed in an aqueous solution, and iron oxide is synthesized in the pores not covered by the wax on the bilateral asymmetric spheres of the mesopores. After synthesizing the iron oxide, the wax is again dissolved in hexane to obtain a submicron bilateral asymmetric magnetic sphere. The immunologically active bilateral asymmetric particles obtained in this preparation are referred to as A-3.

Preparation Example 4 - Preparation method of bilateral asymmetric particles having immunologically active, magnetic and fluorescent molecules

The preparation method of this preparation example is similar to the preparation method of the above Preparation Examples 2 and 3, and the main feature is that 1/2 number of bilateral asymmetric particles will have a fluorescence resonance energy transfer donor, such as Marina Blue, and the remaining 1/2 number The bilateral asymmetric particles will have a fluorescent resonance energy transfer acceptor such as 6-(7-Nitrobenzofurazan-4-ylamino)hexanoic acid. In addition, one of the bilateral asymmetric particles having a fluorescent resonance energy transfer donor and the bilateral asymmetric particles having a fluorescent resonance energy transfer acceptor will have iron oxide, and the other will not have iron oxide. The bilateral asymmetric particles prepared in this preparation example are referred to as A-4.

Preparation Example 5 - Preparation method of immunologically active bilateral asymmetric particles

The preparation method of the preparation example is similar to the preparation method of the above preparation example 1. The main difference is that the preparation step does not perform the step of attaching the fluorescent dye, so that the bilateral asymmetric particles obtained have no fluorescence resonance energy transfer characteristics. , referred to as A-5.

Method of forming a tetrahedral polymeric structure

Specific Example 1 - Taking hepatitis B virus as an example

The bilateral asymmetric particles (A-1) in the above Preparation Example 1 were placed in a solution, and then B-hepatic virus having a diameter of 60 to 70 nm was slowly poured into a solution containing bilateral asymmetric particles (A-1). In order to perform a common co-assembly process, a tetrahedral polymerization structure, referred to as B-1, is formed. It should be noted that the ratio of the bilateral asymmetric particles (A-1) to the virus must be much larger than 4:1, for example, 30:1, so as to ensure that all viruses can be bilaterally asymmetric particles (A -1) coated.

Specific example 2, 3 - taking a simulated virus as an example

The bilateral asymmetric particles (A-1, A-3) in the above Preparation Examples 1 and 3 were placed in a solution, and then the simulated virus having a diameter of 60 to 70 nm was slowly poured into a bilateral asymmetric particle (A). In the solution of -1, A-3), a co-assembly process is carried out to form a tetrahedral polymerization structure, abbreviated as B-2 and B-3. Among them, the simulated virus is a surface carboxylated polyacrylonitrile nanobead (CH470, commercial polymer sphere) having a particle diameter of 80 nm and having a molecule of 1-(3-dimethylaminopropyl) on its surface. N-(3-Dimethylaminopropyl-N'-ethylcarbodiimide hydrochloride (EDC) is attached to biotin, and biotin is used as an antigen. Polyacrylonitrile nanobeads have been pre-loaded with the fluorescent dye Chromeon 470. In other words, the fluorescent dye Marina Blue in the bilateral asymmetric particles (A-1) acts as a donor for the fluorescence resonance energy transfer, while the fluorescent dye Chromeeon 470 in the polyacrylonitrile nanobeads is the fluorescent resonance. Receptor substance for energy transfer. When the distance between the donor substance and the acceptor substance is close enough, for example, 1-10 nm, the phenomenon of fluorescence resonance energy transfer can be observed.

Specific Example 4 - Taking hepatitis B virus as an example

The bilateral asymmetric particles (A-2) in the above Preparation Example 2 were placed in a solution, and then the hepatitis B virus having a diameter of 60 to 70 nm was slowly poured into a solution containing bilateral asymmetric particles (A-2). In order to perform a co-assembly process, a tetrahedral polymerization structure, referred to as B-4, is formed. It should be noted that the ratio of bilateral asymmetric particles (A-2) to virus must be much greater than 4:1, for example 30:1, so that all viruses can be bilaterally asymmetric particles (A). -2) coated. Furthermore, most of the tetrahedral polymeric structures (B-4) consist of at least one bilateral asymmetric particle with a fluorescent resonance energy transfer donor (Marina Blue) and at least one fluorescent resonance. Bilateral asymmetric particles of the energy transfer receptor (6-(7-Nitrobenzofurazan-4-ylamino) hexanoic acid). Preferably, the tetrahedral polymeric structure (B-4) is composed of two bilateral asymmetric particles having a fluorescent resonance energy transfer donor and two bilateral asymmetric particles having a fluorescent resonance energy transfer acceptor.

Specific Example 5 - Taking hepatitis B virus as an example

The bilateral asymmetric particles (A-4) in the above Preparation Example 4 were placed in a solution, and then the hepatitis B virus having a diameter of 60 to 70 nm was slowly poured into a solution containing bilateral asymmetric particles (A-4). In order to perform a common co-assembly process, a tetrahedral polymerization structure, referred to as B-5, is formed. It should be noted that the ratio of the bilateral asymmetric particles (A-4) to the virus must be much larger than 4:1, for example, 30:1, so as to ensure that all viruses can be bilaterally asymmetric particles (A -2) coated. Furthermore, most of the tetrahedral polymeric structures (B-5) consist of at least one bilateral asymmetric particle with a fluorescent resonance energy transfer donor (Mlarina Blue) and at least one with fluorescence resonance. Bilateral asymmetric particles of the energy transfer receptor (6-(7-Nitrobenzofurazan-4-ylamino) hexanoic acid). Preferably, the tetrahedral polymeric structure (B-5) is composed of two bilateral asymmetric particles having a fluorescent resonance energy transfer donor and two bilateral asymmetric particles having a fluorescent resonance energy transfer acceptor.

Method for detecting biomolecules

Test Example 1 - Detection of tetrahedral polymerization structure by electron microscopy

For the electron microscopic examination of the tetrahedral polymerization structure (B-1) of the specific example 1, please refer to Fig. 5, which proves that the four immunologically active bilateral asymmetric particles are simultaneously bonded to the single virus by the biomolecules of the respective surfaces. The tetrahedral structure is formed, and the accommodating space formed in the middle of the tetrahedron can capture a single virus of a specific diameter (particle size is between 50 and 100 nm, especially between 80 and 100 nm), so that the subsequent quantitative detection is more accurate. The space that can be accommodated in the center of the tetrahedral structure is approximately equal to the volume of the virus particles of 100 nm in diameter.

Test Example 2 - Detection of tetrahedral polymerization structure by particle size analysis

The particle size analysis was carried out for the tetrahedral polymerization structure (B-1) of Specific Example 1. Please refer to Figure 6. When the ratio between the number of bilateral asymmetric particles and B-hepatic virus is greater than 10, no more signals of hepatitis B virus will appear. Therefore, it can be determined that the hepatitis B virus can be completely immunologically active. Captured by symmetric particles. In addition, referring to Figure 7, the curve formed by code L27-31-06 represents the average particle size of the tetrahedron and excess bilateral asymmetric particles in the solution. The average particle size of the curve is from tetrahedron and single bilateral asymmetric particles. The proportion of the number is determined. By analyzing the distribution of the curve, the amount of B-hepatic virus in the solution can be pushed back.

Test Example 3 - Detection of tetrahedral polymerization structure by fluorescence resonance energy transfer

The amount of bilateral asymmetric particles (A-1) was fixed in solution, and different amounts of Chromeon 470 (simulated virus) were added, from 0 wt% to 3.5 wt%, to form a polymer structure having different tetrahedrons (B-2). Concentration of solution. Among them, the simulated virus corresponds to the simulated virus in the above specific example 2. Next, FRET detection was performed on each mixed solution, and the results are shown in Fig. 8. Referring to Fig. 8, the FRET luminescence signal of 611 nm was observed, and it was found that the signal intensity was increased with the increase of the concentration of Chromeo 470 (simulated virus). Increase in proportion. Therefore, with the help of the appropriate calibration curve, we can reverse the number of Chromene 470, that is, the number of simulated viruses, from the FRET illuminating signal intensity at the test end.

Test Example 4 - Detection of tetrahedral polymer structure by fluorescence resonance energy transfer

FRET detection was performed on the tetrahedral polymerization structure (B-4) of Specific Example 4. Since most of the hepatitis B virus can be simultaneously treated by two bilateral asymmetric particles with a fluorescence resonance energy transfer donor (Marina Blue) and two fluorescent resonance energy transfer receptors (6-(7-Nitrobenzofurazan-4-ylamino) The bilateral asymmetric particles of hexanoic acid are coated. When two adjacent bilateral asymmetric particle systems in each tetrahedral polymer structure are less than 10 nm, the corresponding fluorescence resonance energy transfer donor will resonate with the fluorescence. The energy transfer receptor produces a FRET signal. In other words, the amount of hepatitis B virus can be reversed by measuring the FRET luminescence signal.

Test Example 5 - Detection of tetrahedral polymerization structure by fluorescence resonance energy transfer and application of magnetic field

FRET detection was performed on the tetrahedral polymerization structure (B-3) of Specific Example 3. The detection method of this test example is similar to the detection method of the above test example 4, and the main difference between the two is that the test example 5 further utilizes a magnetic generating device such as a magnet or an electromagnet to generate a magnetic field to attract a tetrahedral polymer structure dispersed in the solution. (B-3), the tetrahedral polymeric structure (B-3) is concentrated in a specific region in the solution, for example, concentrated near the side wall of the container. By focusing on the tetrahedral polymeric structure (B-3), the signal strength of FRET can be effectively enhanced. Referring to Fig. 9, the signal intensity at a specific wavelength (611 nm) is increased by 13.4 times by applying a magnetic field as compared with the case where no magnetic field is applied.

It should be noted that since the bilateral asymmetric particles (A-3) have only one of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor. Therefore, even if the magnetic field simultaneously attracts magnetic bilateral asymmetric particles (A-3) (or free magnetic bilateral asymmetric particles) that do not capture any simulated virus, the distance between the free magnetic bilateral asymmetric particles is reduced to 1 to 10 nm. These free magnetic bilateral asymmetric particles will not emit any FRET signal (that is, no noise will be generated), which will affect the quantitative results of the virus.

Test Example 6 - Detection of tetrahedral polymerization structure by fluorescence resonance energy transfer

FRET detection was performed on the tetrahedral polymerization structure (B-5) of Specific Example 5. The detection method of this test example is similar to the detection method of the above test example 4, and the main difference between the two is that the test example 5 further utilizes a magnetic generating device such as a magnet or an electromagnet to generate a magnetic field to attract a tetrahedral polymer structure dispersed in the solution. (B-5), the tetrahedral polymeric structure (B-5) is concentrated in a specific region in the solution, for example, concentrated near the side wall of the container. By concentrating the tetrahedral polymeric structure (B-5), the signal intensity of FRET can be effectively enhanced by at least 2-3 times.

It should be noted that one of the bilateral asymmetric particles (A-4) having a fluorescent resonance energy transfer donor and the bilateral asymmetric particles (A-4) having a fluorescent resonance energy transfer acceptor will have Iron oxide, while the other does not have iron oxide, that is, bilateral asymmetric particles that can be attracted by the magnetic field will only have the same kind of fluorescent molecules (ie, fluorescent resonance energy transfer donor or fluorescent resonance energy transfer) One of the bodies). Therefore, even if the magnetic field simultaneously attracts magnetic bilateral asymmetric particles (A-4) (or free magnetic bilateral asymmetric particles) that do not capture any hepatitis B virus, the distance between the free magnetic bilateral asymmetric particles is reduced to 1~ At 10 nm, these free magnetic bilateral asymmetric particles will not emit any FRET signal (ie, no noise will be generated), which will affect the quantitative results of the virus.

Test Example 7 - Centrifugal separation of tetrahedral polymerization structure

The tetrahedral polymeric structures of the foregoing specific examples 1, 2, 3, 4, and 5 (B-1, B-2, B-3, B-4, B) compared to a single bilateral asymmetric particle and a single side object -5) It has a large density, so that the tetrahedral polymerization structure can be quickly concentrated and collected by a rotary centrifugal separation device, thereby judging the number of sides to be side.

Compared with the prior art, the bilateral asymmetric particles and self-assembled polymers provided by the embodiments of the present invention can be used for sensitive, rapid, low-cost and accurate quantitative detection of biomolecules. In addition, the tetrahedral polymerized structure of one embodiment of the present invention has a larger density than the bilateral asymmetric particles and the side to be side, so that it can be quickly concentrated and collected by the centrifugal separation device to determine the number of objects to be side. Moreover, an embodiment of the present invention applies a magnetic field to a magnetic tetrahedral polymeric structure, which can not only rapidly aggregate a magnetic tetrahedral polymeric structure, but also enhance the corresponding fluorescence resonance if measured by the fluorescence resonance energy transfer intensity. The signal strength of the energy transfer greatly improves the efficiency and accuracy of the detection. In summary, the invention can be widely applied to clinical screening for rapid screening of viruses, clinical invasive medical products and even for detecting water micro-viruses.

Claims (33)

A bilateral asymmetric particle (Janus particle) comprising: a low-dimensional substrate, the surface of the low-dimensional substrate comprises a biomolecule modified region; and a plurality of biomolecules fixed to the surface of the low-dimensional substrate and Located in the modified region of the biomolecule, and the free end of each biomolecule has specific binding to the microorganism, wherein the area of the modified region of the biomolecule and the area of the low-dimensional substrate have the following relationship: (1) /5) AS≦AB≦(1/2)AS where AB represents the total area of the biomolecule modified region; and AS represents the total area of the low dimensional substrate. The bilateral asymmetric particles according to claim 1, wherein the low-dimensional substrate is a spherical substrate, a columnar substrate or a dumbbell-shaped substrate. The bilateral asymmetric particles of claim 1, wherein the low-dimensional substrate is a spherical substrate having a medium pore. The bilateral asymmetric particle according to claim 1, wherein the biomolecule modification region is a continuous region and is disposed on one side of the low dimensional substrate. The bilateral asymmetric particles according to claim 1, wherein the biomolecules are uniformly distributed in the modified region of the biomolecule. The bilateral asymmetric particles of claim 1, wherein each of the biomolecules is an antibody. The bilateral asymmetric particle according to claim 1, wherein the surface of the low-dimensional substrate further comprises a non-bio-modified region, wherein the non-bio-modified region is a continuous region and is disposed on One side of the low dimensional substrate. The bilateral asymmetric particles according to claim 7, wherein the area of the non-bio-modified region is represented by the following formula: AM = AS-AB wherein AM represents the total area of the non-bio-modified region. The bilateral asymmetric particles according to claim 7, wherein a plurality of magnetic substances are further included in the non-bio-modified region. The bilateral asymmetric particles of claim 9, wherein the low-dimensional substrate further comprises a plurality of holes, and the magnetic substances are disposed in the holes. The bilateral asymmetric particle according to claim 1, wherein the bilateral asymmetric particle further comprises a plurality of fluorescent molecules fixed to the surface of the low-dimensional substrate and located in the modified region of the biomolecule. The bilateral asymmetric particles of claim 11, wherein each of the fluorescent molecules is a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor or a single fluorescent molecule. A tetrahedral polymeric structure comprising: a microorganism having a surface comprising a plurality of biomolecules; and four bilateral asymmetric particles surrounding the microorganism, wherein each of the bilateral asymmetric particles comprises: a low dimensional substrate The surface of the low-dimensional substrate comprises a biomolecule modified region; and a plurality of other biomolecules are located in the modified region of the biomolecule, wherein the two ends of each of the other biomolecules are respectively fixed to the low Dimensional substrate surface and the microorganism Each of the biomolecules on the surface of the body. The tetrahedral polymeric structure according to claim 13, wherein the area of the modified region of the biomolecule and the area of the low-dimensional substrate have the following relationship: (1/5) AS≦AB≦ (1/2) AS wherein AB represents the total area of the biomolecule modified region; and AS represents the total area of the low dimensional substrate. The tetrahedral polymeric structure according to claim 13, wherein each of the biomolecules on the surface of the low-dimensional substrate has a specific binding force to each of the biomolecules located on the surface of the microorganism. The tetrahedral polymeric structure of claim 13, wherein each of the bilateral asymmetric particles further comprises a plurality of first fluorescent molecules, each of the first fluorescent molecules being fixed to the surface of the low-dimensional substrate In the biomolecule modification region, the microorganism further comprises a plurality of second fluorescent molecules, each of the second fluorescent molecules being fixed to the surface of the microorganism, and each of the first fluorescent molecules is a fluorescent resonance energy transfer One of the donor or fluorescent resonance energy transfer acceptors, each of the second fluorescent molecules being the other of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor. The tetrahedral polymeric structure according to claim 13, wherein one of the bilateral asymmetric particles further comprises a plurality of first fluorescent molecules, each of the first fluorescent molecules being fixed to the low dimension In the biomolecule modified region of the surface of the substrate, the other of the bilateral asymmetric particles further includes a plurality of second fluorescent molecules, each of the second fluorescent molecules being fixed to the surface of the low-dimensional substrate In the biomolecule modification region, each of the first fluorescent molecules is fluorescent One of a vibrational energy transfer donor or a fluorescent resonance energy transfer acceptor, each of the second fluorescent molecules being the other of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor. The tetrahedral polymeric structure according to claim 17, wherein the surface of each of the low-dimensional substrates further comprises a non-bio-modified region, the non-bio-modified region being a continuous region and disposed on the low-dimensional basis One side of the material. The tetrahedral polymeric structure according to claim 18, wherein the bilateral asymmetric particles having the first fluorescent molecules fixed on the surface further comprise a plurality of magnetic substances, the non-living disposed on the low-dimensional substrate The bilateral asymmetric particles having the second fluorescent molecules immobilized on the surface do not include any magnetic substance in the modified region. A method for fabricating bilateral asymmetric particles, comprising: providing at least one low-dimensional substrate; adsorbing the low-dimensional substrate on a surface of a fibrous network structure; performing a heating process to cause the low-dimensional substrate portion Sinking into the fibrous network structure to form a depressed portion, and projecting a portion of the low-dimensional substrate to the surface of the fibrous network structure to form a protrusion, wherein the area of the protrusion is low The ratio of the area of the dimensional substrate is between 0.2 and 0.5; forming a surface modifying layer on the surface of the protruding portion; forming a protective layer on the surface modifying layer; forming the surface modifying layer and the protective layer Thereafter, the low dimensional substrate is detached from the surface of the fibrous network structure; after the low dimensional substrate is detached from the surface of the fibrous network structure, the protective layer is removed; A biomolecule layer is disposed on the surface modifying layer, wherein the biomolecule layer is fixed to the surface modifying layer. The method for producing a bilateral asymmetric particle according to claim 20, wherein the fiber network structure is composed of at least one electrospun fiber, and the segments of the electrospun fiber are stacked and wound with each other. The method for producing a bilateral asymmetric particle according to claim 20, wherein the low-dimensional substrate is a mesoporous spherical substrate comprising a plurality of holes. A method for detecting biomolecules, comprising: providing a plurality of bilateral asymmetric particles, each of the bilateral asymmetric particles comprising: a low-dimensional substrate, the surface of the low-dimensional substrate comprising a biomolecule modified region; and a plurality of living organisms The molecules each include a fixed end and a free end, each of the fixed ends being fixed to the surface of the low-dimensional substrate and located in the biomolecule modified region, wherein the area of the biomolecule modified region and the low portion The area of the dimensional substrate has the following relationship: (1/5) AS≦AB≦(1/2)AS where AB represents the total area of the modified region of the biomolecule; and AS represents the total of the low-dimensional substrate Area; providing a microorganism comprising a plurality of other biomolecules disposed on a surface of the microorganism; and mixing the bilateral asymmetric particles and the microorganism with each other such that the bilateral asymmetry is located The free ends of each of the biomolecules on the surface of the particles are attached to each of the biomolecules on the surface of the microorganism. The method of detecting biomolecules according to claim 23, wherein the low-dimensional substrate is a low-dimensional substrate of a mesopores. The method for detecting a biomolecule according to claim 23, wherein the diameter of the low-dimensional substrate and the diameter of the microorganism have the following relationship: 0.15 ≦ (VD/D) ≦ 0.3, wherein D represents the low dimension The diameter of the substrate; and VD represents the diameter of the microorganism. The method for detecting a biomolecule according to claim 23, wherein the biomolecule modification region is a continuous region and is disposed on one side of the low-dimensional substrate. The method for detecting biomolecules according to claim 23, wherein the biomolecules disposed on the surface of the low-dimensional substrate are uniformly distributed in the biomolecule-modified region. The method for detecting a biomolecule according to claim 23, wherein each of the biomolecules on the surface of the low-dimensional substrate has a specific binding force to each of the biomolecules on the surface of the microorganism. The method for detecting biomolecules according to claim 23, wherein each of the bilateral asymmetric particles further comprises a plurality of first fluorescent molecules, each of the first fluorescent molecules being fixed to the surface of the low-dimensional substrate In the modified region of the biomolecule, the microorganism further includes a plurality of second fluorescent molecules, each of the second fluorescent molecules being fixed to a surface of the microorganism, and each of the first fluorescent molecules is a fluorescent resonance One of energy transfer donors or fluorescent resonance energy transfer receptors Each of the second fluorescent molecules is the other of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor. The method for detecting a biomolecule according to claim 23, wherein a part of the particles of the bilateral asymmetric particles further comprises a plurality of first fluorescent molecules, each of the first fluorescent molecules being fixed to the low dimension a surface of the substrate and located in the modified region of the biomolecule, and the other particles in the bilateral asymmetric particles further comprise a plurality of second fluorescent molecules, each of the second fluorescent molecules being fixed to the low-dimensional substrate And the surface of the biomolecule is in the modified region, and each of the first fluorescent molecules is one of a fluorescent resonance energy transfer donor or a fluorescent resonance energy transfer acceptor, and each of the second fluorescent molecules is a fluorescent The other of the optical resonance energy transfer donor or the fluorescent resonance energy transfer acceptor. The method for detecting biomolecules according to claim 30, wherein the surface of each of the low-dimensional substrates further comprises a non-bio-modified region, wherein the non-bio-modified region is a continuous region and is disposed at the low dimension. One side of the substrate. The method for detecting a biomolecule according to claim 31, wherein each of the bilateral asymmetric particles having the first fluorescent molecules immobilized on the surface further comprises a plurality of magnetic substances disposed on the low-dimensional substrate. In the non-bio-modified region, each of the bilateral asymmetric particles having the second fluorescent molecules immobilized on the surface does not include any magnetic substance. The method for detecting biomolecules according to claim 30, wherein the number of the bilateral asymmetric particles on which the first fluorescent molecules are fixed on the surface is the same as the surface on which the second fluorescent molecules are immobilized. The number of these bilateral asymmetric particles.