CN111374151A - Preparation method of multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition - Google Patents
Preparation method of multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition Download PDFInfo
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- CN111374151A CN111374151A CN202010194827.6A CN202010194827A CN111374151A CN 111374151 A CN111374151 A CN 111374151A CN 202010194827 A CN202010194827 A CN 202010194827A CN 111374151 A CN111374151 A CN 111374151A
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Abstract
The invention discloses a preparation method of a multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition, which comprises the following steps: 1) preparing amino-modified tannic acid TAA, wherein the structural formula of the TAA is as follows:wherein the content of the first and second substances,2) dissolving the TAA of the step 1) in the tris (hydroxymethyl) aminomethaneSoaking the Ti sheet or the SPR chip in a buffer solution to obtain Ti-TAA or Au-TAA; the concentration of TAA is 1-10 mg/ml. 3) Soaking the Ti-TAA or AU-TAA obtained in the step 2) in a water solution of silver nitrate, succinimide-polyethylene glycol-N-hydroxysuccinimide or biotin polyethylene glycol active ester to obtain a material with the functions of antibiosis, anti-protein adsorption or specific recognition, wherein the concentration of the Ti-TAA or AU-TAA is 1-10 mg/ml. Has the functions of anti-protein adsorption with bactericidal effect and specificity recognition.
Description
Technical Field
The invention belongs to the field of antibacterial and anti-protein adsorption materials, and relates to a preparation method of a multifunctional material containing TAA for sterilization, anti-protein adsorption and specific recognition.
Background
The mussel inspiring surface chemistry mimics sticky mussel byssus protein 5(Mefp-5) with broad surface coating potential. Catecholamines and their derivatives are very important for mussel inspiring surface chemistry. Dopamine, one of catecholamine derivatives, can be self-polymerized under alkaline conditions, and forms a universal coating on the surfaces of various substrates through a simple dip coating process. In addition to surface-adhesive and film-forming capabilities, Polydopamine (PDA) also has chemical reactivity, metal-chelating and reducing capabilities, and is easily post-surface functionalized. However, the darker color of PDA coatings limits their widespread use in many areas.
Polyphenols consist of high contents of dihydroxyphenyl (catechol) and trihydroxyphenyl (pyrogallol) groups and are found in coffee, wine, tea, chocolate, fruits, vegetables, etc. They have good biological activity, such as anti-tumor, anti-oxidation, anti-allergy, anti-diabetes, anti-inflammatory, antibacterial and antiviral, etc. Chemical and physical properties are also provided to the functional material by physical interactions, metal coordination and borate complexation, and covalent bonds. Due to the surface adhesion properties of dihydroxyphenyl and trihydroxyphenyl groups, polyphenols are used to build functional surfaces and to adjust surface properties. However, the deposition of Tannic Acid (TA) requires a high salt concentration (0.6M), which limits its practical application.
The development of TA-based surface chemistry or functionalization strategies is very important to broaden its potential application area. Inspired by the unique chemical structure and self-polymerization properties of dopamine, primary amine groups were introduced into the gallic acid moiety in TA by Williamson ether synthesis. Biotin polyethylene glycol active ester (Biotin-PEG-NHS) and succinimide-polyethylene glycol-N-hydroxysuccinimide (mPEG-NHS) were grafted onto the TAA coating by NHS ester-amine coupling reaction for specific recognition or anti-protein adsorption. TAA coatings can be used to reduce silver ions in situ to form Ag NPs, and the resulting Ag NPs-loaded TAA coatings can inhibit bacterial adhesion and prevent biofilm formation.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for preparing a multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition, wherein the prepared multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition has good antibacterial effect, protein adsorption resistance and specific recognition effect.
In order to achieve the purpose, the invention provides the following technical scheme:
1. a preparation method of a multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition comprises the following preparation steps: 1) preparing amino-modified tannic acid TAA, wherein the structural formula of the TAA is as follows:
2) dissolving the TAA obtained in the step 1) in a tris buffer solution, and soaking a Ti sheet or an SPR chip in the buffer solution to obtain titanium Ti-TAA or Au-TAA; the concentration of the TAA is 1-10 mg/ml.
3) Soaking the Ti-TAA or AU-TAA obtained in the step 2) in a water solution of silver nitrate, succinimide-polyethylene glycol-N-hydroxysuccinimide or biotin polyethylene glycol active ester to obtain a material with antibacterial, anti-protein adsorption or specific recognition functions, wherein the concentration of the Ti-TAA or AU-TAA is 1-10 mg/ml.
Further, the step 3) is to soak the Ti-TAA in the step 2) in a silver nitrate water solution for 10-20 hours to obtain Ti-TAA-Ag NPs.
Further, the step 3) is to soak the Au-TAA in the step 2) in a water solution of succinimide-polyethylene glycol-N-hydroxysuccinimide for 20-30 hours to obtain Au-TAA-PEG.
Further, the step 3) is to soak the Ti-TAA or AU-TAA obtained in the step 2) in a biotin polyethylene glycol active ester aqueous solution for 20-30 hours to obtain Au-TAA-biotin.
Further, the concentrations of the aqueous solution of silver nitrate, succinimide-polyethylene glycol-N-hydroxysuccinimide or biotin polyethylene glycol active ester in the step 3) are 1-100 mg/ml, 1-50 mg/ml and 1-50 mg/ml respectively.
Further, the preparation steps of the TAA in the step 1) are as follows:
a) dissolving tannic acid in N, N-dimethylformamide, and sequentially adding N-Boc-3-aminopropyl bromide and K2CO3Reacting at 50-70 ℃, cooling to room temperature after the reaction is finished, adding deionized water, mixing uniformly, adding an HCl aqueous solution, acidifying, extracting with ethyl acetate, combining organic layers, washing with deionized water, drying, filtering, removing a solvent, dissolving a residue in ethanol, dialyzing, and drying to obtain TA-Boc-amino;
b) dissolving the TA-Boc-amino obtained in the step a) in ethanol, introducing hydrogen chloride gas, and reacting to obtain amino-modified tannic acid TAA.
Further, in the step a), 38.3-38.8 parts of tannic acid, 22-22.8 parts of N-Boc-3-aminopropyl bromide and 38.4-39.7 parts of sodium carbonate are calculated according to parts by weight.
Further, the reaction in the step a) is carried out for 4-10 hours, and the drying agent is Na2SO4。
Further, the reaction time of the TA-Boc-amino and the hydrogen chloride in the step b) is 4-10 hours.
The invention has the beneficial effects that: the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition, which is prepared by the invention, not only has a good sterilization effect, but also has good protein adsorption resistance and specific recognition functions. The method has the advantages of simple reaction steps, mild reaction conditions, low risk and the like.
Drawings
In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 is a preparation route diagram of a multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition.
FIG. 2 is a graph showing the survival rate of Ti-TAA-Ag NPs bactericidal material on the Ti surface of Escherichia coli and Staphylococcus aureus.
FIG. 3 is an SEM image of Ti-TAA-Ag NPs bactericidal material on the surface of Ti against Escherichia coli and Staphylococcus aureus.
FIG. 4 is a fluorescence microscopic image showing the bacterial adhesion and biofilm formation of Ti-TAA-Ag NPs bactericidal material to Staphylococcus aureus.
FIG. 5 is a SPR graph showing adsorption of the Au-TAA-PEG anti-protein adsorption material to bovine serum albumin and fibrinogen.
FIG. 6 is an SPR graph formed by adsorption of the Au-TAA-biotin specific recognition material avidin and a histogram of the adsorbed avidin content.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
The Ti-TAA-Ag NPs, Au-TAA-PEG, Au-TAA-biotin of the present invention
The material is prepared by the reaction shown in figure 1, a titanium sheet is taken as a substrate, amino-modified tannic acid and silver nanoparticles are coated on the titanium sheet, an SPR chip is taken as a substrate, amino-modified tannic acid and PEG or Biotin are coated on the substrate, the amino-modified tannic acid is prepared by removing a Boc protecting group of tannic acid-Boc-amino from hydrogen chloride gas, the tannic acid-Boc-amino is prepared by taking potassium carbonate as an acid-binding agent and reacting tannic acid with N-Boc-3-aminopropyl bromide, and the prepared material is used for performing sterilization effect, anti-protein adsorption and specificity identification tests. In the invention, staphylococcus aureus (S.aureus) and escherichia coli (E.coli) are selected as representatives of gram-positive bacteria and gram-negative bacteria and are used for researching the bactericidal action of the Ti-TAA-Ag NPs material; selecting Bovine Serum Albumin (BSA) and Fibrinogen (FBG) as non-specific protein representatives for researching the protein adsorption resistance of the Au-TAA-PEG material; avidin (avidin) was selected for the study of the specific recognition effect of the Au-TAA-biotin material.
Example 1
Preparing a Ti-TAA-Ag NPs bactericidal material:
1) 20g of tannic acid was dissolved in 50mL of N, N-dimethylformamide, and 11.7g N-Boc-3-aminopropylbromide and 20.3g K were added in this order2CO3. The reaction mixture was stirred under argon at 60 ℃ for 6 hours. After cooling to room temperature, 100mL of deionized water was added and mixed well, the solution was acidified by the addition of aqueous HCl (6M) and extracted three times with 100mL of ethyl acetate. The organic layers were combined, washed twice with 100mL deionized water, and Na2SO4And (5) drying. After filtration, the solvent was removed by rotary evaporation. The residue was dissolved in 50mL of ethanol and dialyzed against ethanol for two days. Removing the solvent by rotary evaporation, and drying under reduced pressure to obtain the TA-Boc-amino as a white solid;
2) dissolving 10g of tannic acid-Boc-amino in the step 1) in 100mL of ethanol, and introducing hydrogen chloride gas to react for 6 hours to obtain amino-modified tannic acid TAA;
3) dissolving 80mg of TAA obtained in the step 2) in 40mL of Tris buffer solution, soaking the titanium sheet or the SPR chip in the solution for 24 hours, washing and drying to obtain titanium sheet Ti-TAA with the amino-modified tannic acid coating;
4) soaking the Ti-TAA in the step 3) in 40mL of AgNO3And (C ═ 17mg/mL) in an aqueous solution for 12 hours, washing and drying to obtain titanium sheet Ti-TAA-Ag NPs coated with silver nanoparticles.
Example 2
Preparing an Au-TAA-PEG protein-resistant adsorption material:
1) 20g of tannic acid was dissolved in 50mL of N, N-dimethylformamide and 11.7g N-Boc-3-aminopropylbromide and 20.3g K2CO3 were added in this order. The reaction mixture was stirred under argon at 60 ℃ for 6 hours. After cooling to room temperature, 100mL of deionized water was added and mixed well, the solution was acidified by the addition of aqueous HCl (6M) and extracted three times with 100mL of ethyl acetate. The organic layers were combined, washed twice with 100mL deionized water, and dried over Na2SO 4. After filtration, the solvent was removed by rotary evaporation. The residue was dissolved in 50mL of ethanol and dialyzed against ethanol for two days. Removing the solvent by rotary evaporation, and drying under reduced pressure to obtain the TA-Boc-amino as a white solid;
2) dissolving 10g of tannic acid-Boc-amino in the step 1) in 100mL of ethanol, and introducing hydrogen chloride gas to react for 6 hours to obtain amino-modified tannic acid TAA;
3) dissolving 80mg of TAA in the step 2) in 40mL of Tris buffer solution, soaking the SPR chip in the buffer solution for 24 hours, washing and drying to obtain the amino-modified tannin coated chip Au-TAA;
4) and (3) soaking the Au-TAA in the step 3) in 40mL of an aqueous solution of succinimide-polyethylene glycol-N-hydroxysuccinimide (mPEG-NHS) (C is 1mg/mL) for 24 hours, washing and drying to obtain the TAA and PEG coated SPR chip Au-TAA-PEG.
Example 3
Preparing an Au-TAA-biotin specific recognition material:
1) 20g of tannic acid was dissolved in 50mL of N, N-dimethylformamide, and 11.7g N-Boc-3-aminopropylbromide and 20.3g K were added in this order2CO3. The reaction mixture was stirred under argon at 60 ℃ for 6 hours. After cooling to room temperature, 100mL of deionized water was added and mixed well, the solution was acidified by the addition of aqueous HCl (6M) and extracted three times with 100mL of ethyl acetate. The organic layers were combined, washed twice with 100mL deionized water, and dried over Na2SO 4. After filtration, the solvent was removed by rotary evaporation. The residue was dissolved in 50mL of ethanol and dialyzed against ethanol for two days. Removing the solvent by rotary evaporation, and drying under reduced pressure to obtain the TA-Boc-amino as a white solid;
2) dissolving 10g of tannic acid-Boc-amino in the step 1) in 100mL of ethanol, and introducing hydrogen chloride gas to react for 6 hours to obtain amino-modified tannic acid TAA;
3) dissolving 80mg of TAA in the step 2) in 40mL of Tris buffer solution, soaking the SPR chip in the buffer solution for 24 hours, washing and drying to obtain the amino-modified tannin coated chip Au-TAA;
4) and (3) soaking the Au-TAA in the step 3) in 40mL of Biotin polyethylene glycol active ester (Biotin-PEG-NHS) (C is 1mg/mL) aqueous solution for 24 hours, washing and drying to obtain the SPR chip Au-TAA-Biotin coated with the Biotin-tannic acid.
Example 4
The method for quantitatively detecting the sterilization effect by using the sterilization material comprises the following specific contents:
1. and (3) culturing bacteria: the freezing tube containing the strain was removed from the-20 ℃ refrigerator, thawed, and then inoculated into Tryptone Soya Broth (TSB) medium and shake-cultured at 37 ℃ for future use. The culture is generally carried out one day in advance.
2. Quantitative detection of the sterilization effect comprises the following specific steps (the following equipment and liquid are sterilized in advance):
1) the strain was first diluted to 1 × 10 in PBS buffer7;
2) Putting blank titanium sheets with the specification of 1cm × 1cm, Ti-TAA and Ti-TAA-Ag NPs into a 24-pore plate, respectively adding 1mL of bacterial solution, and culturing at 37 ℃ for 24 hours;
3) pipetting 1mL of PBS buffer solution, adding the PBS buffer solution into a 24-well plate, carefully washing off bacteria which are not adsorbed on the titanium plate, and repeating the steps for three times;
4) respectively marking 15mL of centrifuge tubes, adding 3mL of PBS buffer solution into the centrifuge tubes, putting the washed titanium sheets into the corresponding centrifuge tubes, and carrying out ultrasonic treatment for 5 minutes;
5) respectively marking 1.5mL centrifuge tubes, adding 900 mu L PBS buffer solution into the centrifuge tubes, sucking 100 mu L solution after ultrasonic treatment, and performing gradient dilution;
6) sucking 100 μ L of the diluted solution, adding to TSB agar plate, coating with coating rod until the bacteria solution is completely absorbed, culturing at 37 deg.C for 24 hr, and counting;
FIG. 2 is a graph showing the survival numbers of Escherichia coli and Staphylococcus aureus on the Ti surface of the sterilization material, (a) a colony graph obtained by adsorbing Escherichia coli and Staphylococcus aureus on the Ti surface and diluting the bacteria by 1000 or 100 times, and (b) the numbers of bacteria adsorbed on the Ti surface by Escherichia coli and Staphylococcus aureus, wherein the numbers of the live Escherichia coli and Staphylococcus aureus on the blank Ti are about 2.0 × 106And 1.9 × 105Is smallCell/cm2The number of live E.coli and S.aureus on the Ti-TAA1 surface was reduced to 53% and 60% respectively on the blank Ti surface, the number of live E.coli and S.aureus on the Ti-TAA-Ag NPs was reduced to 1.0 × 105And 2.7 × 103Individual cell/cm2。
Example 5
The sterilization effect qualitative detection is carried out by using the sterilization material, and the specific contents are as follows:
1. and (3) culturing bacteria: the freezing tube containing the strain was removed from the-20 ℃ refrigerator, thawed, and then inoculated into Tryptone Soya Broth (TSB) medium and shake-cultured at 37 ℃ for future use. The culture is generally carried out one day in advance.
2. The qualitative detection of the sterilization effect comprises the following specific steps of (sterilizing the following equipment and liquid in advance):
1) the strain was first diluted to 1 × 10 in PBS buffer7;
2) Putting blank titanium sheets with the specification of 1cm × 1cm, Ti-TAA and Ti-TAA-Ag NPs into a 24-pore plate, respectively adding 1mL of bacterial solution, and culturing at 37 ℃ for 24 hours;
3) pipetting 1mL of PBS buffer solution, adding the PBS buffer solution into a 24-well plate, carefully washing out bacteria which are not adsorbed on the titanium plate, and repeating twice;
4) sucking 1mL of 3% glutaraldehyde solution, adding into a 24-pore plate, and soaking overnight at 4 ℃;
5) sucking out the glutaraldehyde solution, sucking 1mL of ethanol solution, adding the ethanol solution into a 24-pore plate, and performing gradient dehydration for 10 minutes each time, wherein the concentration is 25%, 50%, 75% and 100% respectively;
FIG. 3 is a scanning electron micrograph of the bacterial adhesion of the biocidal material to Escherichia coli and Staphylococcus aureus, and FIGS. 3 a-c and FIGS. d-f show the adhesion of Escherichia coli and Staphylococcus aureus to blank and modified Ti, respectively. As shown in the figure, a large number of bacteria adhered to the surface of the blank Ti, and a little less bacteria adhered to the surface of the Ti-TAA than to the surface of the blank Ti. There were fewer bacteria on the surface of the Ti-TAA-Ag NPs. It can be shown that Ti-TAA-Ag NPs can effectively inhibit bacterial adhesion.
Example 6
The sterilization effect qualitative detection is carried out by using the sterilization material, and the specific contents are as follows:
1. and (3) culturing bacteria: the freezing tube containing the strain was removed from the-20 ℃ refrigerator, thawed, and then inoculated into Tryptone Soya Broth (TSB) medium and shake-cultured at 37 ℃ for future use. The culture is generally carried out one day in advance.
2. The qualitative detection of the sterilization effect comprises the following specific steps of (sterilizing the following equipment and liquid in advance):
1) the strain is diluted to 1 × 10 by TSB and PBS buffer (1: 1)5;
2) Putting blank titanium sheets with the specification of 1cm × 1cm, Ti-TAA and Ti-TAA-Ag NPs into a 24-pore plate, respectively adding 1mL of bacterial solution, and culturing at 37 ℃ for 24 hours;
3) pipetting 1mL of PBS buffer solution, adding the PBS buffer solution into a 24-well plate, carefully washing out bacteria which are not adsorbed on the titanium plate, and repeating twice;
4) pipetting 15 mu L of a combined dye of SYTO9 and ammonium iodide (PI) to the titanium plate, and incubating for 15 minutes;
5) sucking 1mL of distilled water, adding the distilled water into a 24-well plate, and carefully washing off the fluorescent dye which is not adsorbed on the titanium plate;
FIG. 4 is a fluorescence microscopic image of bacterial adhesion and biofilm formation of the bactericidal material against Staphylococcus aureus: live cells were stained fluorescent green with SYTO9 dye and dead cells were stained fluorescent red with PI dye. As shown in the figure, the bacteria adhered to the surface of the blank Ti emitted strong green fluorescence with a small amount of red fluorescence, and the bacteria adhered to the surface of Ti-TAA were less than those on the surface of the blank Ti, and the red fluorescence was more prominent than the green fluorescence. Ti-TAA-Ag NPs have negligible green and red fluorescence on their surface. The Ti-TAA-Ag NPs can effectively inhibit bacterial adhesion and biofilm formation.
Example 7
The detection of the protein-resistant adsorption effect by using the Au-TAA-PEG protein-resistant adsorption material comprises the following specific contents:
1. the method for detecting the protein adsorption effect comprises the following specific steps:
1) dissolving two proteins of BSA and FBG in PBS buffer solution to reach the concentration of 200 mu g/mL;
2) placing the blank SPR chip, Au-TAA and Au-TAA-PEG into an SPR tester;
3) passing PBS buffer through the SPR chip at a flow rate of 20 μ L/min to obtain a stable baseline;
4) add 200. mu.L BSA or FBG solution through the well and flow at 20. mu.L/min for 10 min;
5) the sensing surface was rinsed with PBS at the same flow rate to remove loosely adsorbed proteins.
FIG. 5 is a SPR graph showing the effect of the Au-TAA-PEG anti-protein adsorption material on the anti-protein adsorption of BSA and FBG proteins: a is an SPR graph of the protein adsorption effect of the protein adsorption material on BSA protein, and b is an SPR graph of the protein adsorption effect of the protein adsorption material on FBG protein, wherein the larger the numerical value is, the higher the content of the adsorbed protein is. As shown in the figure, a large amount of protein is adhered to the surfaces of the blank SPR chip and the Au-TAA, and the difference is not shown between the blank SPR chip and the Au-TAA-PEG, and the contents of BSA and FBG protein adsorbed on the Au-TAA-PEG surface are respectively reduced to 45% and 53% of the blank SPR chip. It can be shown that Au-TAA-PEG can inhibit the adsorption of nonspecific protein.
Example 8
The specific recognition effect detection is carried out by using the Au-TAA-biotin specific recognition material, and the specific contents are as follows:
1. detecting the specificity recognition effect, which comprises the following steps:
1) dissolving three proteins of BSA, FBG and avidin in PBS buffer solution until the concentration is 200 mu g/mL;
2) placing the Au-TAA-biotin into an SPR tester;
3) passing PBS buffer through the SPR chip at a flow rate of 20 μ L/min to obtain a stable baseline;
4) adding 200. mu.L of BSA, FBG or avidin solution through the well, and flowing at a flow rate of 20. mu.L/min for 10 minutes;
5) the sensing surface was rinsed with PBS at the same flow rate to remove loosely adsorbed proteins.
6) Before passing through protein adsorptionThe change in post-SPR angular shift was used to calculate the amount of protein adsorbed. Typically, an angular change of 0.1 ° corresponds to 1ng/mm on the sensing surface2The adsorbed protein of (4);
FIG. 6 is a graph showing the protein adsorption effect of the Au-TAA-biotin anti-protein adsorption material on three proteins, namely BSA, FBG and avidin: a is an SPR image of the specific recognition material adsorbing three proteins of BSA, FBG and avidin, and b is a content column diagram of the specific recognition material adsorbing three proteins of BSA, FBG and avidin. As shown in the figure, the adsorption amounts of the Au-TAA-biotin specific recognition material to two nonspecific proteins of BSA and FBG are respectively 2.10 +/-0.68 and 4.74 +/-1.33 ng/mm2And adsorption to avidin is 41.02 +/-9.21 ng/mm2. The Au-TAAA-biotin can be obtained, not only can effectively inhibit the adsorption of nonspecific protein, but also has strong identification capability on avidin.
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (9)
1. A preparation method of a multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition is characterized by comprising the following preparation steps:
1) preparing amino-modified tannic acid TAA, wherein the structural formula of the TAA is as follows:
2) dissolving the TAA obtained in the step 1) in a tris buffer solution, and soaking a Ti sheet or an SPR chip in the buffer solution to obtain titanium Ti-TAA or Au-TAA; the concentration of the TAA is 1-10 mg/ml.
3) Soaking the Ti-TAA or AU-TAA obtained in the step 2) in a water solution of silver nitrate, succinimide-polyethylene glycol-N-hydroxysuccinimide or biotin polyethylene glycol active ester to obtain a material with antibacterial, anti-protein adsorption or specific recognition functions, wherein the concentration of the Ti-TAA or AU-TAA is 1-10 mg/ml.
2. The preparation method of the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition according to claim 1, wherein the step 3) is to soak the Ti-TAA in the step 2) in a silver nitrate aqueous solution for 10-20 hours to obtain Ti-TAA-Ag NPs.
3. The method for preparing the multifunctional material containing TAA with sterilization, protein adsorption resistance and specific recognition functions according to claim 1, wherein the step 3) is to soak the Au-TAA in the step 2) in an aqueous solution of succinimide-polyethylene glycol-N-hydroxysuccinimide for 20-30 hours to obtain Au-TAA-PEG.
4. The method for preparing the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition according to claim 1, wherein the step 3) is to soak the Ti-TAA or AU-TAA obtained in the step 2) in the biotin polyethylene glycol active ester aqueous solution for 20-30 hours to obtain Au-TAA-biotin.
5. The preparation method of the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition according to claim 1, wherein the concentrations of the aqueous solution of silver nitrate, succinimide-polyethylene glycol-N-hydroxysuccinimide or biotin polyethylene glycol active ester in step 3) are 1-100 mg/ml, 1-50 mg/ml and 1-50 mg/ml, respectively.
6. The method for preparing the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition according to claim 1, wherein the TAA prepared in step 1) comprises the following steps:
a) dissolving tannic acid in N, N-dimethylformamide, and sequentially adding N-Boc-3-aminopropyl bromide and K2CO3Reacting at 50-70 ℃, cooling to room temperature after the reaction is finished, adding deionized water, mixing uniformly, adding an HCl aqueous solution, acidifying, extracting with ethyl acetate, combining organic layers, washing with deionized water, drying, filtering, removing a solvent, dissolving a residue in ethanol, dialyzing, and drying to obtain TA-Boc-amino;
b) dissolving the TA-Boc-amino obtained in the step a) in ethanol, introducing hydrogen chloride gas, and reacting to obtain amino-modified tannic acid TAA.
7. The preparation method of the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition as claimed in claim 6, wherein in step a), the multifunctional material comprises, by weight, 38.3-38.8 parts of tannic acid, 22-22.8 parts of N-Boc-3-aminopropyl bromide and 38.4-39.7 parts of sodium carbonate.
8. The method for preparing the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition according to claim 6, wherein the reaction of step a) is performed for 4-10 hours, and the drying agent is Na2SO4。
9. The method for preparing the multifunctional material containing TAA for sterilization, protein adsorption resistance and specific recognition according to claim 6, wherein the reaction time of TA-Boc-amino and hydrogen chloride in step b) is 4-10 hours.
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