Method for manufacturing fiber assemblies providing binding surfaces for biomaterials and fiber assemblies prepared using this method

Abstract

This invention provides a method for manufacturing a fiber assembly providing a binding surface for biomaterials. According to an embodiment of the invention, the fiber assembly providing a binding surface for biomaterials is prepared by the following steps: 1) preparing a fiber assembly containing multiple fibers; and 2) modifying the fiber surface to provide carboxyl groups, which then react with amine groups present in the biomaterial. The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to the invention allows for the easy and high-content introduction of biomaterials into the fiber assembly. Furthermore, it significantly reduces the binding between biomaterials or the attachment of biomaterials through physical adsorption during the introduction process, thereby enabling the use of biomaterials with high precision and reliability in applications utilizing biomaterials. In addition, since the detachment of biomaterials can be minimized or prevented, specific changes caused by biomaterials can be minimized in applications utilizing biomaterials. Therefore, biomaterials fixed to the surface of the fiber assembly according to the invention can be applied in various fields such as materials engineering, bioengineering, and medicine.

Description

Technical Field

[0001] This invention relates to a method for manufacturing fiber assemblies, specifically, to a method for manufacturing fiber assemblies that provide a bonding surface for biomaterials, and to a fiber assembly for preparing a bonding surface for biomaterials using this method. Background Technology

[0002] Researchers are constantly exploring ways to immobilize various biological materials into structures, and structures immobilized with biological materials are widely used in various fields such as materials engineering, life sciences, and medicine.

[0003] Methods for immobilizing biomaterials onto structures have historically relied heavily on physical adsorption or ion bonding based on surface charge differences due to their ease of fabrication. However, this method suffers from a fatal flaw: low binding strength prevents the biomaterials from being stably bonded to the structure, leading to easy detachment.

[0004] On the other hand, porous beads have traditionally been used as structures for immobilizing biological materials, especially magnetic beads which are used in consideration of recycling after use. However, magnetic beads have a small surface area, limiting the amount of biological material they can immobilize.

[0005] Furthermore, there are methods that involve directly coating the surface of a container, which serves as a structure for attaching biomaterials, with the biomaterials for fixation. However, this simple coating fixation often leads to the detachment of the biomaterials. Moreover, depending on the container material used for coating the biomaterials, the coating process may not be easy, potentially accelerating the detachment of biomaterials coated on such materials. Additionally, the detachment of biomaterials can reduce the analytical sensitivity, accuracy, precision, and stability of applications utilizing biomaterials, such as when measuring markers. Furthermore, when biomaterials aggregate randomly on the surface without a specific orientation, spatial constraints can reduce the binding efficiency or binding capacity on the surface. Moreover, the container surface is limited to loading biomaterials onto bare surfaces, thus restricting the amount of biomaterials that can be loaded.

[0006] This will lead to the further research into a structure with a stable and highly integrated bonding surface. Summary of the Invention

[0007] Purpose of the invention

[0008] The purpose of this invention is to solve the problems existing in the prior art and provide a method for manufacturing a fiber assembly that can provide a binding surface for biomaterials, and a fiber assembly that provides a binding surface for biomaterials prepared by the method. This fiber assembly can introduce as much biomaterial as possible, stably fix the introduced biomaterials to prevent detachment, and facilitate the full functioning of the introduced biomaterials.

[0009] Technical solution

[0010] To achieve the above objectives, the present invention provides a method for manufacturing a fiber assembly that provides a binding surface for biomaterials, comprising the following steps: 1) preparing a fiber assembly containing multiple fibers; and 2) modifying the fiber surface to provide carboxyl groups on the fiber surface, thereby reacting with amine groups present in the biomaterial.

[0011] According to one embodiment of the present invention, the fiber comprises one or more mixtures selected from the group consisting of polyvinylidene fluoride, polyacrylonitrile, polyester, polyamide, polylactic acid, polyvinyl alcohol, polystyrene, polyglycolic acid, polyvinyl chloride, polyvinylpyrrolidone, polyurethane and polyethersulfone (PES), or copolymers thereof.

[0012] Furthermore, step 2 may also include the following steps: 2-1) treating the surface of the fiber aggregate with an alkaline solution with a pH of 12.0 or higher; and

[0013] 2-2) The step of treating the surface of the fiber aggregate treated with an alkaline solution with a solution containing a carboxyl compound.

[0014] Furthermore, the carboxyl-containing compound may include compounds containing at least three or more carboxyl groups. In this case, the carboxyl-containing compound may be citric acid.

[0015] In addition, the fiber assembly comprises polyacrylonitrile fibers, and the carboxyl-containing compound may be citric acid.

[0016] In addition, steps 2-1) and 2-2) are each carried out independently at a temperature of 50–70°C for more than 24 hours.

[0017] Furthermore, the solvent for the solution containing the carboxyl compound can be any one or more selected from alcohols having 3 to 10 carbon atoms and water. As an example, the solvent can be a mixture of water and alcohol in a weight ratio of 1:0.2 to 1.8.

[0018] Furthermore, the concentration of the carboxyl-containing compound in the carboxylic acid compound solution can be 8–15 M.

[0019] Furthermore, the present invention provides a fiber assembly for providing a binding surface for biomaterials, characterized in that the fiber assembly is formed by a plurality of fibers present on the surface by means of a manufacturing method according to the present invention, wherein carboxyl groups react with the amine groups present on the biomaterial of the present invention.

[0020] According to one embodiment of the present invention, the fiber comprises polyacrylonitrile fiber, and the carboxyl group may be derived from citric acid.

[0021] Furthermore, the average diameter of the carboxyl group is less than 1 μm.

[0022] In addition, a fiber assembly immobilized with biomaterial is provided, the assembly comprising: a fiber assembly according to the present invention; and having at least one amine group, the amine group being capable of forming a covalent bond with a carboxyl group present on the fiber surface within the fiber assembly, thereby being able to bind biomaterial to the fiber surface.

[0023] According to one embodiment of the present invention, the biomaterial includes any one or more of the following: enzymes containing at least one amino group or modified to have at least one amino group, biological signaling molecules, and biomolecules. The enzyme comprises one or more selected from the group consisting of carbonic acid dehydratase, sugar oxidase, trypsin, chymotrypsin, subtilisin, papain, thermophilic protease, lipase, peroxidase, acyltransferase, lactonease, protease, tyrosinase, laccase, cellulase, xylanase, organic polycarbohydrate hydrolase, cholinesterase, formate dehydrogenase, acetaldehyde dehydrogenase, alcohol dehydrogenase, glucose dehydrogenase, and glucose isomerase; the biological molecule may be one or more selected from the group consisting of chemokines, cytokines, cell survival factors, cell proliferation factors, and cell differentiation factors; the biological molecule may be one selected from the group consisting of albumin, insulin, collagen, antibody, antigen, protein A, protein G, avidin, streptavidin, neutral avidin, biotin, nucleic acid, peptide, phytohemagglutinin, glycosylated protein, cell, and carbohydrate.

[0024] Invention Effects

[0025] The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to the present invention enables the easy and high-content introduction of biomaterials into the fiber assembly. Furthermore, it significantly reduces the binding between biomaterials or the attachment of biomaterials through physical adsorption during the introduction process, thereby enabling the utilization of biomaterials with high precision and reliability in applications utilizing biomaterials. In addition, since the detachment of biomaterials can be minimized or prevented, specific changes caused by biomaterials can be minimized in applications utilizing biomaterials. Therefore, the biomaterials fixed to the surface of the fiber assembly according to the present invention can be applied in various fields such as materials engineering, bioengineering, and medicine. Attached Figure Description

[0026] Figure 1 The FTIR spectra are shown according to an embodiment and a comparative example of the present invention. Detailed Implementation

[0027] The present invention will be described in detail below through embodiments to enable those skilled in the art to readily implement it. The present invention can be implemented in many different forms and is not limited to the embodiments described below.

[0028] The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to the present invention includes: 1) a step of preparing a fiber assembly containing a plurality of fibers; and 2) a step of modifying the fiber surface to have carboxyl groups on the fiber surface, and reacting them with amine groups present in the biomaterial.

[0029] First, the step of preparing a fiber assembly containing multiple fibers is carried out as step 1).

[0030] The function of the fibrous aggregate is to serve as a support for the introduced biomaterial. The biomaterial aggregate can be formed by multiple fibers randomly stacked along the height direction, thereby possessing a 3D network structure and thus a non-uniform surface morphology. The surface of the fibrous aggregate is essentially a collection of the outer surfaces of the individual fibers, thus possessing the advantage of a significantly increased specific surface area for supporting biomaterials compared to non-porous supports of the same volume.

[0031] The fiber assembly can be collectively referred to as a fiber web, and therefore its manufacturing method can be prepared according to the usual fiber web manufacturing method. The present invention does not particularly limit the specific method for manufacturing the fiber assembly.

[0032] However, to achieve a superior specific surface area, the fiber assembly according to an embodiment of the present invention is formed from fibers with an average diameter of 2 μm or less, more preferably less than 1 μm. Fibers with this diameter can be manufactured using conventional methods for producing ultrafine fibers by melt spinning of island-island fibers followed by a weight reduction process, or by electrospinning. When using electrospinning, the fiber assembly can be directly formed on the surface of the containment space within the container used for impregnating biomaterials, eliminating the need for attaching a support to the container, thus simplifying the manufacturing process and shortening manufacturing time.

[0033] The electrospinning can be carried out directly using known electrospinning equipment and conventional electrospinning conditions, or with appropriate modifications. Specifically, the voltage intensity, air gap height, humidity, and temperature applied during electrospinning can be appropriately adjusted according to the material and content of the fiber-forming components contained in the spinning solution, the type of solvent used to dissolve them, and the required fiber diameter.

[0034] Furthermore, the fiber-forming component can be selected from suitable, well-known fiber-forming components, taking into account the required physical properties such as fiber manufacturing method, chemical resistance, mechanical strength, and flexibility. As an example, the fiber-forming component is selected from one polymer compound or a mixture of two or more thereof, or a copolymer formed by copolymerizing two or more thereof, chosen from the group consisting of polyvinylidene fluoride, polyacrylonitrile, polyester, polyamide, polylactic acid, polyvinyl alcohol, polystyrene, polyglycolic acid, polyvinyl chloride, polyvinylpyrrolidone, polyurethane, and polyethersulfone (PES). Preferably, the fiber-forming component is polyacrylonitrile, which has the advantage of allowing for a higher loading of the biomaterials described later onto the fiber aggregate compared to other types of fiber-forming components.

[0035] In addition, the basis weight of the fiber assembly can be, for example, 1 to 100 g / m², and can be adjusted appropriately according to requirements.

[0036] Next, according to step 2 of the present invention, the modification of the fiber assembly is carried out to load carboxyl groups onto the fiber surface.

[0037] The carboxyl group is a binding functional group used to fix biomaterials to the fibrous assembly, and more specifically, a functional group used to form amides with amine groups in the biomaterials.

[0038] The carboxyl groups can be loaded onto the fiber surface using known modification methods. However, it is preferable to include the steps of: 2-1) treating the surface of the fiber assembly with an alkaline solution; and 2-2) treating the surface of the fiber assembly treated with the alkaline solution with a solution containing a carboxyl-containing compound, thereby loading the carboxyl groups onto the fiber surface.

[0039] Step 2-1) involves contacting the surface of the fiber aggregate, specifically the fiber surface, with an alkaline solution, thereby forming hydroxyl groups on the fiber surface. An example of the alkaline solution is a sodium hydroxide solution, with a concentration of 1–20 M, preferably 5–10 M. Furthermore, the alkaline solution has a pH value of 12.0 or higher, preferably 13.0 or higher, and more preferably 13.5 or higher. The alkaline solution treatment method can specifically be impregnation. The alkaline solution treatment temperature can be 50–70°C. The alkaline solution treatment time can be 1 minute to 30 hours, preferably 15 hours or more, and more preferably 24 hours or more. If the alkaline solution treatment time is insufficient, a significant increase in the amount of biomaterial introduced into the fiber aggregate may not be achieved.

[0040] Next, as step 2-2), it can be carried out by treating the surface of the fiber assembly treated with the alkaline solution with a solution containing a carboxyl compound. In this case, a water washing process of the alkaline-treated fiber assembly can be performed before step 2-2), but it is not limited to this.

[0041] The carboxyl-containing compound is a compound containing at least three carboxyl groups. For example, it can be one or more selected from the group consisting of citric acid, acrylic acid-maleic acid copolymer, polyacrylic acid, styrene-sulfonic acid-maleic acid copolymer, and styrene-maleic acid copolymer. If the carboxyl-containing compound contains only two carboxyl groups, there may be a significant reduction in the amount of biomaterial introduced into the fiber aggregate.

[0042] The preferred carboxyl-containing compound is citric acid. Compared to using polymeric compounds such as polyacrylic acid, it has the advantage of increasing the amount of biomaterial introduced, preventing non-specific introduction of biomaterial, and enabling the introduced biomaterial to fully perform its functions.

[0043] The solvent for the solution containing the carboxyl-containing compound can be any one or more of an alcohol or water, selected from straight-chain or branched alcohols with 1 to 10 carbon atoms, preferably 3 to 10 carbon atoms. More preferably, the solvent further includes any one or more of propanol and isopropanol, thereby having the advantage of further increasing the amount of carboxyl groups introduced.

[0044] Furthermore, more preferably, the solvent comprises water and alcohol in a weight ratio of 1:0.2 to 1.8. This minimizes solution vaporization during the extended reaction time in step 2-2), thus providing the advantage of more stably loading carboxyl groups onto the fiber surface. Additionally, the concentration of the carboxyl compound in the solution can be 1 to 20 M, preferably 8 to 15 M, allowing for sufficient loading of carboxyl groups while ensuring the full functionality of the introduced biomaterial. If the concentration of the carboxyl compound in the solution exceeds the preferred range and is too high, the increase in the amount of loaded carboxyl groups is minimal. When the biomaterial has multiple amine groups, one biomaterial may bind to multiple carboxyl groups, leading to problems such as biomaterial deformation or functional impairment.

[0045] Furthermore, step 2-2) can be carried out at a temperature of 50-70°C for 1 minute to 30 hours, preferably for 15 hours or more, and more preferably for 24 hours or more. If the treatment and reaction time of the solution containing carboxyl compounds are insufficient, the amount of biomaterial introduced into the fibrous aggregate cannot be significantly increased.

[0046] On the other hand, this invention uses a fiber surface modification method to expose carboxyl groups on the surface, which, compared to methods that add carboxyl-containing compounds to the spinning solution for spinning, has the advantage of significantly increasing the amount of biomaterial bound to the fiber aggregate. This is a highly specific effect, entirely different from the ability to predict results from FT-IR data of carboxyl-containing compounds. Specifically, refer to... Figure 1 It can be seen that Comparative Example 1, in which a fiber assembly containing a carboxyl compound is spun in a spinning solution, has a higher surface modification value than Example 1, in which the fiber assembly is treated with an alkaline solution and then treated with a solution containing a carboxyl compound for surface modification. The carboxyl group is located at 1740 cm. -1 The peak value at that location is significantly higher. However, as can be seen from Table 1 described later, the amount of biotin corresponding to the amount of streptomycin introduced in Comparative Example 1, which is an example of biomaterial, is less than 1 / 100th the amount of avidin introduced in Example 1. This is contrary to the result that could be predicted by FT-IR data. Therefore, it can be seen that the cellulose aggregate according to the present invention has the ability to introduce a very large amount of biomaterial.

[0047] Furthermore, the present invention can realize a fiber assembly that provides a binding surface for biomaterials through the above manufacturing method, which is formed by fibers loaded with carboxyl groups on the surface, the carboxyl groups being able to react with amine groups contained in the biomaterials.

[0048] In this case, the fiber preferably comprises polyacrylonitrile fiber, and the carboxyl group may be derived from citric acid, which enables the introduction of biomaterials into the fiber assembly in a significantly increased amount.

[0049] Furthermore, the present invention also provides a bio-assembly immobilized with biomaterial, comprising: a fiber assembly according to the present invention; and a biomaterial having at least one amine group, said amine group being capable of forming a covalent bond with a carboxyl group present on the fiber surface within the fiber assembly, thereby binding to the biomaterial on the fiber surface.

[0050] The biomaterial can be a known substance used in materials engineering, bioengineering, or medicine; it can be a substance present in or absent from a living organism, as long as it is usable in biological reactions. The biomaterial can be organic or inorganic; examples of organic materials include carbohydrates, proteins, nucleic acids, lipids, or low-molecular-weight forms of these substances. An example of the biomaterial can be any one or more of the following: modified enzymes containing at least one amino group, biological signaling molecules, and biomolecules.

[0051] The enzyme can be any known enzyme without particular limitation. Specific examples include one or more enzymes selected from the group consisting of carbonic acid dehydratase, sugar oxidase, trypsin, chymotrypsin, subtilisin, papain, thermophilic protease, lipase, peroxidase, acyltransferase, lactonease, protease, tyrosinase, laccase, cellulase, xylanase, organic polycarbohydrate hydrolase, cholinesterase, formate dehydrogenase, acetaldehyde dehydrogenase, alcohol dehydrogenase, glucose dehydrogenase, and glucose isomerase.

[0052] Furthermore, the biological signaling molecule can be any known one without particular limitation, for example, it can be one of the group consisting of chemokines, cytokines, cell survival factors, cell proliferation factors and cell differentiation factors.

[0053] In addition, the biomolecule may be one or more selected from the group consisting of albumin, insulin, collagen, antibody, antigen, protein A, protein G, avidin, streptavidin, neutral avidin, biotin, nucleic acid, peptide, phytohemagglutinin, glycosylated protein, cell and carbohydrate.

[0054] In addition, the biomaterial may also contain additional markers capable of recognizing the biomaterial. These markers may be fluorescent substances, fluorescent substance-binding proteins, luminescent substances, luminescent substance-binding proteins, or enzymes, etc. Appropriate known markers may be selected, and the present invention does not impose any particular limitation on them.

[0055] Furthermore, the biomaterial can be incorporated into the fiber aggregate by known methods, for example, by EDC or EDC / NHS coupling reaction.

[0056] The fiber assemblies that provide a binding surface for biomaterials according to the present invention can be applied to various reagent kits, devices, biosensors, supports for cell or tissue culture, bio-batteries, and many other applications used in detection, diagnosis, and analysis, and can be widely used in various industries.

[0057] Example

[0058] The present invention will be further described in detail through the following embodiments. It should be understood that the following embodiments are not intended to limit the scope of the present invention, but are provided to help understand the present invention.

[0059] Example 1

[0060] Polyacrylonitrile (PAN) nanofibers with an average diameter of 300 μm and a basis weight of 30 g / m² were prepared. The prepared nanofibers were subjected to alkaline treatment by immersing them in a 5 M sodium hydroxide aqueous solution at 60 °C for 24 hours. Subsequently, the alkaline-treated nanofibers were immersed in a propanol solution containing 10 M citric acid at 60 °C for 24 hours to prepare a fiber aggregate with carboxyl groups loaded on the fiber surface.

[0061] Example 2

[0062] The fiber aggregate with carboxyl groups loaded on the fiber surface was prepared in the same manner as in Example 1, except that a polyvinylidene fluoride nanofiber mesh with an average diameter of 500 μm and a basis weight of 30 g / m² was used instead of the fiber aggregate.

[0063] Example 3

[0064] The process was the same as in Example 1, except that a polyurethane / polyvinylidene fluoride nanofiber web with an average diameter of 600 μm and a basis weight of 30 g / m² was used instead of the original nanofiber web to prepare a fiber aggregate with carboxyl groups loaded on the fiber surface. In this case, the nanofiber web can be prepared using a spinning solution of polyurethane and polyvinylidene fluoride mixed in a 5:5 weight ratio.

[0065] Comparative Example 1

[0066] To prepare the spinning solution, 36g of polyacrylonitrile was first added to 114.8g of dimethylacetamide / 49.2g of acetone and stirred at 80°C for 6 hours with a magnetic stir bar to dissolve and prepare a mixed solution. Next, after cooling the collected mixed solution to room temperature, 1.8g of citric acid was mixed into 200mL of the mixed solution to prepare the spinning solution. The prepared electrospinning solution was placed in a solution tank and fed at a rate of 15μl / min / hole to prepare a nanofiber web aggregate with an average diameter of 300μm and a basis weight of 25g / m². At this time, the temperature in the spinning zone was 28°C, the humidity was maintained at 40%, and the distance between the collector and the spinning nozzle was 18cm. Then, following the same method as in Example 1, the prepared fiber aggregate was treated with a solution containing an alkaline solution to prepare a fiber aggregate with carboxyl groups loaded on the fiber surface.

[0067] Comparative Example 2

[0068] Prepared using the same method as Comparative Example 1, but with polyvinylidene fluoride instead of polyacrylonitrile in the spinning solution, a nanofiber web assemblies with an average particle size of 500 μm and a basis weight of 30 g / m² were prepared, thereby preparing a fiber assembly with surface-loaded carboxyl groups.

[0069] Comparative Example 3

[0070] Prepared using the same method as Comparative Example 1, a nanofiber web aggregate with an average particle size of 500 μm and a basis weight of 30 g / m² was prepared by using a mixture of polyurethane and polyvinylidene fluoride in a weight ratio of 3:7 instead of polyacrylonitrile in the spinning solution. This resulted in the preparation of a fiber aggregate with surface-loaded carboxyl groups.

[0071] Experimental Example 1

[0072] The surface-loaded carboxyl fiber assemblies prepared in Examples 1 to 3 and Comparative Examples 1 to 3 were used to prepare test pieces with a width and length of 1 cm and 5 cm, respectively. The test pieces were then immersed in 15 mM sodium methyl ester sulfonate (MES) at pH 6. In the reaction mixture, 25 mg / 2.5 ml of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (dissolved in MES) was added. After reacting for approximately 30 minutes, the supernatant was removed, and 4 mg / 5 ml of StAv (dissolved in MES) was added to introduce streptavidin (StAv) into the fiber aggregates. Following elution with pH 2 elution buffer, the cells were washed three times with 5 ml of PBST. The binding affinity was evaluated using FITC-Biotin (ThermoFisher) according to the manufacturer's instructions and experimental protocol, and the results are shown in Table 1 below.

[0073] Reagent test kit:

[0074] Table 1

[0075]

[0076] As can be confirmed by Table 1, compared with the fiber assemblies of the comparative example, which contain carboxyl-containing compounds in the spinning solution to load carboxyl groups on the surface, the fiber assemblies of the embodiments have the ability to introduce biomaterials in very excellent amounts.

[0077] Example 4

[0078] The same method as in Experimental Example 1 was used to manufacture the fiber aggregates with surface-loaded carboxyl groups, except that the alkaline solution treatment time was changed to 1 minute.

[0079] Comparative Example 4

[0080] Polyacrylonitrile (PAN) nanofibers were prepared using the same method as in Example 1, without any modification.

[0081] Experiment Example 2

[0082] The FT-IR spectra of the fiber assemblies of Examples 1, 4, Comparative Examples 1 and 4 were confirmed using a Raman spectrometer (LabRamARAMIS IR2), and the results are shown below. Figure 1 middle.

[0083] Reference Figure 1 It can be confirmed that Comparative Example 1, in which a fiber assembly containing a carboxyl-containing compound is spun in a spinning solution, shows a higher surface modification at 1740 cm relative to the carboxyl group compared to Example 1, in which the fiber assembly is treated with an alkaline solution followed by surface modification with a solution containing a carboxyl-containing compound. -1 The peak value at that location is significantly higher.

[0084] However, as can be confirmed from Table 1, the amount of biotin corresponding to the amount of streptomycin introduced in Comparative Example 1, which is a biological material, is less than 1 / 100 of that in Example 1. This is contrary to the result that can be predicted by FT-IR data. Therefore, it can be seen that the cellulose aggregate according to the present invention has the ability to introduce a very large amount of biological material.

[0085] Example 5

[0086] The same method as in Example 1 was used, except that a polyacrylonitrile (PAN) nanofiber mesh with an average diameter of 300 μm and a basis weight of 30 g / m² was used instead of the fiber aggregate, the concentration of the alkali solution was changed to a 10 M sodium hydroxide solution, and the treatment time of the alkali solution and the solution containing carboxyl compounds was changed to 8 hours, thereby preparing a fiber aggregate with carboxyl groups loaded on the fiber surface.

[0087] Example 6

[0088] The same method as in Example 1 was used, except that a polyacrylonitrile (PAN) nanofiber network with an average diameter of 300 μm and a basis weight of 30 g / m² was used instead of the fiber aggregate, the concentration of the alkaline solution was changed to a 10 M sodium hydroxide solution, and the treatment time of the alkaline solution and the solution containing carboxyl compounds was changed to 24 hours, thereby preparing a fiber aggregate with carboxyl groups loaded on the fiber surface.

[0089] Experimental Example 3

[0090] Fiber aggregates immobilized with streptavidin were prepared using the same method as in Experimental Example 1. The binding capacity was evaluated using the same method, and the results are shown in Table 2 below.

[0091] Table 2

[0092]

[0093] As can be confirmed by Table 2, Example 6, which was treated with alkaline solution and carboxyl compound solution for 24 hours, introduced a significantly greater amount of biomaterial compared to Example 5. In terms of the amount introduced per unit treatment time, it can be seen that Example 6 prepared a fiber aggregate with a significantly greater binding amount than that predicted from Example 5, which is more than 3 times greater.

[0094] The above description illustrates one embodiment of the present invention. The purpose of the present invention is not limited to the embodiments described in the specification. Those skilled in the art should understand that other embodiments can be easily conceived by adding, modifying, deleting, or supplementing constituent elements within the scope of the present invention, and these also fall within the protection scope of the present invention.

Claims

1. A method for manufacturing a fiber assembly providing a binding surface for biomaterials, characterized in that, include: 1) The steps for preparing a fiber assembly containing multiple fibers; and 2) The step of modifying the fiber surface to have carboxyl groups on the fiber surface, which then react with the amine groups present in the biomaterial. Step 2) includes the following steps: 2-1) The step of treating the surface of the fiber aggregate with an alkaline solution of pH 12.0 or higher; and 2-2) The step of treating the surface of a fiber assembly treated with an alkaline solution with a solution containing a carboxyl compound having citric acid and using an alcohol having 3 to 10 carbon atoms or an aqueous solution of the alcohol as a solvent to provide carboxyl groups on the fiber surface.

2. The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to claim 1, wherein, The fiber comprises one or more of the following: polyvinylidene fluoride, polyacrylonitrile, polyester, polyamide, polylactic acid, polyvinyl alcohol, polystyrene, polyglycolic acid, polyvinyl chloride, polyvinylpyrrolidone, polyurethane, and polyethersulfone (PES), or copolymers thereof.

3. The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to claim 1, wherein, The fiber assembly comprises polyacrylonitrile fibers, and the carboxyl-containing compound is citric acid.

4. The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to claim 1, wherein, Steps 2-1) and 2-2) are each carried out independently at a temperature of 50–70°C for more than 24 hours.

5. The method for manufacturing a fiber assembly providing a binding surface for biomaterials according to claim 1, wherein, The concentration of the carboxyl-containing compound in the solution is 8–15 M.

6. A fiber assembly formed of a plurality of fibers, providing a binding surface for the biomaterial, by means of the manufacturing method according to any one of claims 1 to 5, wherein carboxyl groups that react with amine groups present in the biomaterial are loaded onto the surface.

7. The fiber assembly according to claim 6, wherein, The fiber includes polyacrylonitrile fiber, and the carboxyl group is derived from citric acid.

8. The fiber assembly according to claim 6, wherein, The average diameter of the fiber is less than 1 μm.

9. A fibrous aggregate immobilized with biomaterials, wherein, It includes the fiber assembly according to claim 6; and has at least one amine group, which is capable of forming a covalent bond with a carboxyl group on the surface of the fiber within the fiber assembly, thereby enabling it to bind to biomaterials on the fiber surface.

10. The biological aggregate with immobilized biological material according to claim 9, wherein, The biomaterials include any one or more of the following: enzymes containing at least one amino group or modified to have at least one amino group; biological signaling molecules; and biomolecules. The enzyme comprises one or more selected from the group consisting of carbonic acid dehydrase, sugar oxidase, lipase, peroxidase, acyltransferase, lactonease, protease, tyrosinase, laccase, cellulase, xylanase, organic polycarbonate hydrolase, cholinesterase, formate dehydrogenase, acetaldehyde dehydrogenase, alcohol dehydrogenase, glucose dehydrogenase, and glucose isomerase. The biological signaling molecules may be cytokines.

11. The biological aggregate with immobilized biological material according to claim 10, wherein the protease comprises: One or more of trypsin, chymotrypsin, subtilisin, papain, and thermophilic protease.

12. The biological aggregate with immobilized biological material according to claim 10, wherein the cytokines include any one or more of chemokines, cell survival factors, cell proliferation factors, and cell differentiation factors.

13. The biological aggregate with immobilized biological material according to claim 10, wherein the biomolecule is selected from the group consisting of albumin, insulin, and collagen.

14. The biological aggregate immobilized with biological material according to claim 10, wherein the biomolecules are antibodies and / or antigens.

15. The biological aggregate immobilized with biological material according to claim 10, wherein the biomolecule is protein A and / or protein G.

16. The biological aggregate immobilized with biological material according to claim 10, wherein the biomolecule is avidin and / or biotin.

17. The biological aggregate with immobilized biological material according to claim 16, wherein the avidin is streptavidin and / or neutral avidin.

18. The biological aggregate immobilized with biological material according to claim 10, wherein the biomolecules are nucleic acids and / or peptides.

19. The biological aggregate with immobilized biological material according to claim 10, wherein the biomolecule is lectin.

20. The biological aggregate with immobilized biological material according to claim 10, wherein the biomolecule is a glycosylated protein.

21. The biological aggregate immobilized with biological material according to claim 10, wherein the biomolecule is a cell.

22. The biological aggregate with immobilized biological material according to claim 10, wherein the biomolecule is a carbohydrate.

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