CN112126405A

CN112126405A - Novel bionic adhesion material and preparation method and application thereof

Info

Publication number: CN112126405A
Application number: CN202011014133.6A
Authority: CN
Inventors: 薛斌; 王天阔; 李英; 曹毅; 秦猛; 王炜
Original assignee: Nanjing Research Institute Of Nanjing University
Current assignee: Nanjing Research Institute Of Nanjing University
Priority date: 2020-09-24
Filing date: 2020-09-24
Publication date: 2020-12-25

Abstract

The invention relates to a novel bionic adhesion material and a preparation method and application thereof, wherein a dopa-containing polypeptide nanofiber is self-assembled to form an intelligent adhesion material; the intelligent polypeptide nanofiber material containing dopa has very excellent biological adhesion performance, can be widely applied to the fields of biomedicine, tissue engineering, intelligent soft materials and the like, is suitable for capturing and releasing various biological cells, and further performs research work on research and development of various biological materials and medicines.

Description

Novel bionic adhesion material and preparation method and application thereof

Technical Field

The invention relates to a novel bionic adhesion material and a preparation method and application thereof.

Background

Single cell analysis and cell therapy have been widely explored and applied in disease diagnosis, cancer treatment, and new drug development. For example, single cell analysis reveals diversity in cell behavior and provides richer data than the commonly used analysis of cell collections; the detection and isolation of Circulating Tumor Cells (CTCs) is of great importance for the diagnosis and treatment of cancer. Thus, the capture and release of cells is of great importance for the diagnosis and treatment of cells. Much effort has been made to develop efficient cell capture and release methods. In general, cell adhesion methods based on surface modification sensitive to pH, temperature, voltage and light have been developed as a means of controlling cell adhesion to surfaces. For example, a temperature sensitive poly (N-isopropylacrylamide) (PNIPAAm) hydrogel coating is used to capture target cells, which are then released by varying the temperature. Other researchers have also achieved this goal by using photo-stimuli responsive polymeric materials containing azobenzene. However, the biocompatibility or capture efficiency of these methods has not been satisfactory for practical use. Therefore, developing effective cell capture and release methods remains challenging.

In recent years, bioadhesives have received much attention due to their strong adhesion strength under water and high biocompatibility. 3, 4-Dihydroxyphenylalanine (DOPA) is a molecule widely used by marine organisms for underwater adhesion and is by far one of the most attractive bioadhesive molecules. The dopa-containing material exhibits excellent adhesion to various materials due to strong interaction forces on inorganic and organic surfaces due to coordination bonds, hydrogen bonds, and hydrophobic interactions. Furthermore, recent studies have also shown that the self-assembly of dopa-containing molecules can greatly improve adhesion energy. The highly efficient adhesion and biocompatibility of dopa makes it well suited for cell capture. However, the adhesion strength of individual dopa molecules is weak and most dopa adhesion is permanent and uncontrollable, limiting the application in cell capture. Thus, the introduction of a synergistic enhancement of adhesion and a moderate release mechanism into the adhesion design of dopa-containing materials is a key factor in cell capture and release.

Disclosure of Invention

Aiming at the defects of the prior art, the invention solves the problems that: provides a novel bionic adhesive material with high-strength adhesive property, a preparation method and application thereof.

In order to solve the problems, the technical scheme adopted by the invention is as follows:

a novel bionic adhesive material is formed by self-assembling dopa-containing polypeptide nano fibers; the polypeptide nanofiber comprises a self-assembly structure polypeptide sequence and an adhesion group; the self-assembly structure polypeptide sequence forms a nanofiber structure through hydrophobic interaction self-assembly; the adhesion group relies on catechol structure to achieve adhesion.

Further, the self-assembly structure polypeptide sequence is 2-naphthylacetic acid-glycine-phenylalanine (2-NapGFF); the adhesion group is a dopa amino acid; the self-assembly structure polypeptide sequence and the adhesion group form a short peptide containing four amino acids through dehydration condensation.

Further, the adhesion material is self-assembled in PBS buffer solution to form nano fibers, and the nano fibers are further wound and combined to form a fiber network structure.

A preparation method of a novel bionic adhesion material comprises the following steps:

s1, modification and protection of dopa molecules: firstly, the anhydrous CH₃CN (23.0mL) was added dihydroxyphenylalanine (Dopa) (2.0g,10.1mmol) and TBDMS-Cl (4.6g,30.5 mmol); cooling with ice water bath for 10min, and adding DBU (4.6mL,30.2mmol) dropwise for more than 10 min; after that, the reaction mixture was allowed to ice-bath for 4h, and then reacted at room temperature for 20 h; after the reaction is finished, removing the solvent by rotary evaporation, adding the mixed product into a silica gel column (the height is 30 cm, the diameter is 2.5 cm), eluting by using a mixed solution of methanol and dichloromethane (gradually increasing from 1:9 to 3:7) to remove impurities, and removing the solvent by rotary evaporation of eluent to obtain a white product Dopa (TBDMS)₂-OH; mixing Dopa (TBDMS)₂OH (8.62g, 31.3mmol) and 40mL 10% Na₂CO₃The solution was added to 113mL THF, which was performed in an ice bath; after the mixture is stirred and reacts for 10 minutes, FmocOSu (6.39g, 19.0mmol) is added, and after the mixture is stirred and reacts for 4 hours in an ice bath, the reaction is continued for 18 hours at room temperature; the solvent was removed by rotary evaporation, and the product mixture was added to 50ml of water and acidified to pH 1 with hydrochloric acid. Then using CH₂Cl₂(3X 50mL) and the extracted CH was extracted₂Cl₂Layer bonding with H₂O (3X 50mL) and saturated brine (50mL) were washed, and anhydrous Na was added₂SO₄Drying, filtering and concentrating the solution, eluting the concentrated product by dichloromethane solution containing 0.5% methanol, and performing rotary evaporation to obtain white powder product Fmoc-Dopa (TBDMS)₂-OH(2.8g，4.4mmol)；

S2, Synthesis of adhesion polypeptide 2-NapGFFDopa: Fmoc-DOPA (TBDMS) obtained in S1₂-OH (1eq.) and DIPEA (4eq.) in CH₂Cl₂(20mL of the resin-1), 2-chlorotrityl chloride resin (1mmol/g, 1eq.) was added thereto, the mixture was stirred at room temperature for 1 hour, and then the solvent was filtered off with 17:2:1 CH₂Cl₂MeOH, DIPEA (v: v: v, 3X 20mL/g resin) washing the reacted resin; then separately using CH₂Cl₂DMF and CH₂Cl₂Thoroughly cleaning the resin, and performing vacuum drying; the product ligation rate was determined to be about 0.5mmol g using 2% DBU/DMF^-1(ii) a The dried resin was swollen in DMF for about 0.5h, filtered to remove the solvent, the Fmoc protecting group was removed by addition of 20% piperidine/DMF (3X 5mL), stirred for 5min, and then separately treated with CH₂Cl₂Washed 3 times with DMF, then the Fmoc protected amino acid (4eq.), HBTU (4eq.), and DIPEA (8eq.) were added to DMF (5mL), mixed with the resin and stirred at room temperature for 2h, then the solvent was filtered, DMF, CH, respectively₂Cl₂And DMF 3 times; the addition sequence of the amino acid protected by Fmoc is Fmoc-Phe-OH, Fmoc-Phe-OH and 2-Nap-Gly-OH respectively; after completion of the reaction, the resin was added to TBAF/THF (15mL, 0.13M) for 45 min to remove the TBDMS protecting group; finally, with 15% TFA/CH₂Cl₂(20mL g^-1Resin) and resin are reacted for 3h at room temperature to cleave off the polypeptide molecule; filtering, evaporating and concentrating the obtained reaction liquid in a rotary manner to remove the solvent, and adding cold ether to precipitate the polypeptide; filtering, collecting, washing with diethyl ether, and vacuum drying to obtain polypeptide product.

Further, the step S1 satisfies the protection of the structure of the DOPAC with TBDMS-Cl to synthesize Dopa (TBDMS)₂OH, Fmoc-Dopa (TBDMS) satisfying protection of the amino terminus by FmocOSu₂-OH。

Further, the step S2 is satisfied with the amino acid addition sequence of Fmoc-Phe-OH, Fmoc-Phe-OH, and 2-Nap-Gly-OH.

The application of a novel bionic adhesion material is applied to high-efficiency adhesion and nonspecific adhesion of biological cells.

The application of the novel bionic adhesion material is applied to the efficient release of biological cells and has no damage to the cells.

The invention has the advantages of

1. Compared with the adhesive material taking other traditional chemical macromolecules as main adhesive components, the adhesive material adopts a novel bionic technology, utilizes biological polypeptide as an adhesive, does not contain any toxic and harmful substances, has excellent biocompatibility and environmental friendliness, and has great effect and value on the research of bioscience.

2. Compared with the adhesion material which is subjected to the de-adhesion by mechanical force or a chemical means with larger irritation or is difficult to de-adhere at all, the adhesion material utilizes the pH response of biomolecules to enable polypeptide molecules to be de-assembled by virtue of small-amplitude pH rise, and the de-adhesion process can be completed. The whole process is convenient, fast and efficient, almost does not threaten cells, and ensures the consistency of the cell life activity before and after adhesion. Has very good practical value for biological research and medicine development.

3. Compared with other adhesive coatings which are adhered by single noncovalent interaction, the adhesive material also overcomes the problems of weak adhesive capacity and low adhesive efficiency of single noncovalent interaction.

4. The adhesion material has the characteristics of simple operation and rapid effect in the process of adhering and releasing cells, and can greatly improve the working efficiency.

5. The adhesive material does not show obvious selectivity for the adhesion of different cells, so the adhesive material has better universality, can be suitable for the adhesion and release of different biological cells, and has wider application range.

Drawings

Fig. 1 is a schematic diagram of the design principle of the novel bionic adhesion material.

FIG. 2 is a characterization map of different polypeptide samples used in the experiment as controls.

FIG. 3 is a characterization map of pH response disassembly of 2-NapGFFDoap.

FIG. 4 is a graph of a single molecular force spectrum experiment for investigating the adhesion energy of the interaction between different polypeptide samples and a glass substrate.

FIG. 5 is a graph of a single molecule force spectrum experiment investigating the adhesion energy of different polypeptide samples and 97h cell interaction.

FIG. 6 is a graph showing the specificity of capture experiments on cells by different polypeptide coatings and the adhesion of 2-NapGFFDopa to different cells by the conventional biological cross-linker, fibrin glue.

FIG. 7 is an experiment of 2-NapGFFDopa releasing cells and an experimental profile of the effect of pH change on cell life activity.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings.

A novel bionic adhesive material is formed by self-assembling dopa-containing polypeptide nano fibers; the polypeptide nanofiber comprises a self-assembly structure polypeptide sequence and an adhesion group; the self-assembly structure polypeptide sequence forms a nanofiber structure through hydrophobic interaction self-assembly; the adhesion group relies on catechol structure to achieve adhesion. The polypeptide sequence of the self-assembly structure is 2-naphthylacetic acid-glycine-phenylalanine (2-NapGFF); the adhesion group is a dopa amino acid; the self-assembly structure polypeptide sequence and the adhesion group form a short peptide containing four amino acids through dehydration condensation. The adhesive material is self-assembled in PBS buffer solution to form nano fibers, and further wound and combined to form a fiber network structure.

s1, modification and protection of dopa molecules: firstly, the anhydrous CH₃CN (23.0mL) was added dihydroxyphenylalanine (Dopa) (2.0g,10.1mmol) and TBDMS-Cl (4.6g,30.5 mmol); cooling with ice water bath for 10min, and adding DBU (4.6mL,30.2mmol) dropwise for more than 10 min; after that, the reaction mixture was allowed to ice-bath for 4h, and then reacted at room temperature for 20 h; after the reaction is finished, removing the solvent by rotary evaporation, adding the mixed product into a silica gel column (the height is 30 cm, the diameter is 2.5 cm), eluting by using a mixed solution of methanol and dichloromethane (gradually increasing from 1:9 to 3:7) to remove impurities, and removing the solvent by rotary evaporation of eluent to obtain a white product Dopa (TBDMS)₂-OH; mixing Dopa (TBDMS)₂OH (8.62g, 31.3mmol) and 40mL 10% Na₂CO₃The solution was added to 113mL THF, which was performed in an ice bath; after the mixture is stirred and reacts for 10 minutes, FmocOSu (6.39g, 19.0mmol) is added, and after the mixture is stirred and reacts for 4 hours in an ice bath, the reaction is continued for 18 hours at room temperature; the solvent was removed by rotary evaporation, and the product mixture was added to 50ml of water and acidified to pH 1 with hydrochloric acid. Then using CH₂Cl₂(3X 50mL) and the extracted CH was extracted₂Cl₂Layer bonding with H₂O (3X 50mL) and saturated brine (50mL) were washed, and anhydrous Na was added₂SO₄Drying, filtering and concentrating the solution, eluting the concentrated product by dichloromethane solution containing 0.5% methanol, and performing rotary evaporation to obtain white powder product Fmoc-Dopa (TBDMS)₂-OH (2.8g, 4.4 mmol); synthesis of Dopa (TBDMS) satisfying the protection of the DOPAC Structure with TBDMS-Cl in step S1₂OH, Fmoc-Dopa (TBDMS) satisfying protection of the amino terminus by FmocOSu₂-OH。

S2, Synthesis of adhesion polypeptide 2-NapGFFDopa: Fmoc-DOPA (TBDMS) obtained in S1₂-OH (1eq.) and DIPEA (4eq.) in CH₂Cl₂(20mL of the resin-1), 2-chlorotrityl chloride resin (1mmol/g, 1eq.) was added thereto, the mixture was stirred at room temperature for 1 hour, and then the solvent was filtered off with 17:2:1 CH₂Cl₂MeOH, DIPEA (v: v: v, 3X 20mL/g resin) washing the reacted resin; then separately using CH₂Cl₂DMF and CH₂Cl₂Thoroughly cleaning the resin, and performing vacuum drying; the product ligation rate was determined to be about 0.5mmol g using 2% DBU/DMF^-1(ii) a The dried resin was swollen in DMF for about 0.5h, filtered to remove the solvent, the Fmoc protecting group was removed by addition of 20% piperidine/DMF (3X 5mL), stirred for 5min, and then separately treated with CH₂Cl₂Washed 3 times with DMF, then the Fmoc protected amino acid (4eq.), HBTU (4eq.), and DIPEA (8eq.) were added to DMF (5mL), mixed with the resin and stirred at room temperature for 2h, then the solvent was filtered, DMF, CH, respectively₂Cl₂And DMF 3 times; the addition sequence of the amino acid protected by Fmoc is Fmoc-Phe-OH, Fmoc-Phe-OH and 2-Nap-Gly-OH respectively; after completion of the reaction, the resin was added to TBAF/THF (15mL, 0.13M) for 45 min to remove the TBDMS protecting group; finally, with 15% TFA/CH₂Cl₂(20mL g^-1Resin) and resin are reacted for 3h at room temperature to cleave off the polypeptide molecule; filtering, evaporating and concentrating the obtained reaction liquid in a rotary manner to remove the solvent, and adding cold ether to precipitate the polypeptide; filtering, collecting, washing with diethyl ether, and vacuum drying to obtain polypeptide product. The sequence of amino acid addition in step S2 was satisfied and was Fmoc-Phe-OH, Fmoc-Phe-OH, and 2-Nap-Gly-OH, respectively.

The novel controllable polypeptide self-assembly nanofiber based on pH sensitivity is used for cell capture and release. By introducing an adhesion group (Dopa) into the self-assembling peptide, the adhesion group is well aligned on both sides of the nanofiber. This unique structure allows the nanofibers to attach to the substrate on one side and the cells on the other side simultaneously. The designed nanofiber has excellent cell capture capacity and is even better than fibrin glue. Furthermore, the captured cells can be efficiently released without being destroyed by pH-induced peptide depolymerization. The adhesive nanofiber has wide application in the fields of biomedicine and tissue engineering. The design principle of such nanofibers also represents a general approach to the design of highly efficient and biocompatible glues.

The invention is inspired by the effect that the high-strength adhesive capacity of marine organism mussel is mainly from dopa contained in byssus protein, and provides a design and preparation idea of a novel adhesive material which has high-efficiency adhesion and release efficiency and is non-toxic and harmless. The main idea is to design and develop a polypeptide nanofiber gel coating containing dopa. As shown in fig. 1, our design is divided into two aspects, namely, an adhesion process, which is performed by using a dopa-containing polypeptide self-assembly molecule to form an entangled nanofiber network structure, and is performed by a series of interactions between dopa molecules distributed on both sides of the fiber and a substrate or a cell. Then, the release process of the cells after adhesion. By changing the pH of the adhesion environment to about 8.4, the disassembly and assembly of the polypeptide fiber network are destroyed, so that the captured cells are released. No chemical substance with any toxic or side effect is used in the whole process, the biocompatibility is very good, the operation is simple and smooth, and the practical efficiency is high.

The following examples of the performance of the present invention were tested:

example 1 morphological and mechanical characterization of the invention.

In the experiments, two additional sets of polypeptide samples were used as controls to verify the principle of excellent adhesion performance of 2-NapGFFDopa. Respectively, 2-NapGFFY containing no dopa and 2-NapGAADopa containing dopa but not capable of self-assembly, as shown in FIG. 2A. Under normal conditions, the tubeThe solid hydrogel product was obtained by rapidly adding a polypeptide solution (DMSO-dissolved) containing 2-napgfdfopa or 2-NapGFFY to phosphate buffer solution (PBS, pH 7.4) and mixing well with shaking. The 2-NapGAADopa is not self-assembled to form a fiber network, and thus the whole is still in a liquid state. Atomic Force Microscopy (AFM) scans also showed very distinct fiber network structures in the 2-NapGFFDopa and 2-NapGFFY samples, as shown in FIGS. 2B-D. The mechanical test by the rheometer can also show that the storage modulus (G') of the 2-NapGFFDopa, the 2-NapGFFY2-NapGFFDopa and the 2-NapGFFY is 1-100 rad s^-1Is higher than the loss modulus (G ") over a wide angular frequency range, indicating that the hydrogels formed by both are physically stable. And the storage modulus of 2-NapGFFDopa is 4 times that of 2-NapGFFY, which indicates that 2-NapGFFDopa has much stronger mechanical strength than 2-NapGFFY, which may be related to the fact that dopa forms hydrogen bonds much more densely than tyrosine. The storage modulus of the corresponding 2-NapGAADopa is smaller than the loss modulus in the low frequency range, indicating that the whole is in a fluid state. As shown in fig. 2E to G. We expect that 2-NapGFFDopa peptide nanofibers will exhibit impressive cell adhesion capability because they have well-aligned cell adhesion groups (Dopa) and gelation of the polypeptide fibers increases the contact area and thus further enhances the adhesion effect.

Example 2 testing of the invention in terms of pH-responsive disassembly.

In terms of pH-responsive disassembly, we used NaHCO₃The pH range is adjusted by the solution. Experiments show that after the pH value of the environment is adjusted from 7.4 to 8.4, the 2-NapGFFDopa hydrogel is gradually changed from a solid state to a liquid state within 30min, and an AFM (atomic force microscope) scanning graph also verifies the disappearance of a fiber network structure. As shown in fig. 3A. This was also confirmed by circular dichroism experiments, where the Circular Dichroism (CD) of the assembled 2-NapGFFDopa exhibited a distinct peak corresponding to the beta sheet structure at 215nm, while no significant signal was observed in the decomposed peptide solution, consistent with AFM imaging results (fig. 3B). Rheological experiments showed that the storage modulus of 2-NapGFFDopa also began to be less than the loss modulus after the pH-raising treatment, also demonstrating the fluidization of the gel (FIG. 3C). These results are shown in the tableObviously, proper pH change can control the dissociation of the 2-NapGFFDopa hydrogel, and is beneficial to the capture and release of cells.

Example 3 the present invention was tested in a single molecule force spectroscopy experiment for the strength of interaction with a substrate.

To study the adhesion performance of the 2-NapGFFDopa fibers, microscale adhesion experiments were performed using the force spectrum of an Atomic Force Microscope (AFM) to determine the adhesion energy, as shown in FIG. 4A. To avoid tip-to-cell damage, a micro glass sphere was attached to the AFM cantilever with epoxy. The glass spheres were lightly pressed down with AFM manipulation to 2-NapGFFDopa hydrogel, so that the glass spheres were coated with 2-NapGFFDopa nanofibers. In a typical force spectroscopy test, the cantilever is at 2000nm s^-1Then held in contact with the substrate for 2s with a contact force of 1nN to allow the polypeptide nanofibers to interact with the substrate and then returned, all experiments being performed in PBS buffer at room temperature. Typical force profile curves for different polypeptides are shown in fig. 4B. The adhesion energy is calculated by integrating the peak area of the force spectrum curve, and histogram analysis is carried out. As shown in FIG. 4C, the adhesion energy of 2-NapGFFDopa (1.3X 10)^-15J) Much larger than 2-NapGFFY (1.11X 10) without dopa^-16J) And inability to self-assemble 2-NapGAADopa (1.21X 10)^-17J) The 2-NapGFFDopa nano-fiber is proved to have stronger adhesion capability, and the difference of the adhesion capability indicates that the self-assembly sequence of the polypeptide and the adhesion group dopa are the key points for effective adhesion. In addition, 2-NapGFFDopa is subjected to NaHCO at pH 8.4₃After the solution treatment, the adhesion energy is obviously reduced, and the important effect of the synergistic effect of the fiber network formed by self-assembly and the Dopa on the adhesion in the polypeptide nano-fiber is further confirmed.

Example 4 the invention was tested in a single molecule force spectroscopy experiment for the strength of interaction with cells.

We analyzed the adhesion strength between polypeptide nanofibers and 97h cells using Atomic Force Microscopy (AFM) force spectroscopy (fig. 5A). A typical force profile of the interaction of polypeptide nanofibers with cells is shown in FIG. 5B. The adhesion energy of each curve was analyzed and counted as in fig. 5C. And interacting with the substrateSimilarly, the adhesion energies of various polypeptides in cell interactions are statistically the same. The adhesion energy of 2-NapGFFDopa is about 1.12X 10^-15J and 2-NapGFFY and 2-NapGAADopa are 7.73X 10, respectively^-16And 5.75X 10^-16J. And as suspected, the adhesion of 2-NapGFFDopa after pH treatment for disassembly decreased dramatically. The high adhesion energy and the rapid decrease in pH-responsive adhesion energy of 2-NapGFFDopa indicate the potential of using this polypeptide nanofiber for controlling cell capture and release.

Example 5 study of the present invention in an experiment for capturing cells.

We demonstrate the feasibility of using 2-NapGFFDopa for cell capture and release by testing the excellent adhesion strength between 2-NapGFFDopa polypeptide nanofibers and polystyrene substrates and cells. We then performed cell capture and release experiments on 2-NapGFFDopa,2-NapGFFY and 2-NapGAADopa coated substrates, and used a polystyrene substrate without coated polypeptide as a blank. In addition, we also used the bioadhesive, fibringlue, commonly used in biological experiments as a reference control to demonstrate the practical utility of 2-NapGFFDopa adherent cells. 97h of cell solution was added to each coating for adsorption for a period of time, followed by washing, and then staining for fluorescence imaging, with specific results shown in FIGS. 6A-E. The number of adherent cells for the different coatings was counted in fig. 6F, while fig. 6G is a statistic of specific adhesion efficiency. It can be seen that 2-NapGFFDopa has a great advantage over other polypeptides, and even shows more efficient adhesion efficiency than the conventional bioadhesive, fibrinlue. FIG. 6H is a test of whether 2-NapGFFDopa specifically adheres to different cells, and the adhesion efficiency of 2-NapGFFDopa to 97H, HK, B1, and 7721 cells was tested. The results show that 2-NapGFFDopa does not show a very significant difference in adhesion efficiency, and has relatively high adhesion efficiency for different cells.

Example 6 investigation of the invention release cells and cell viability assay.

After the cell adhesion experiments were performed, we next needed to perform cell release studies. As shown in FIGS. 7A-B, after pH treatment for disassembly, few cells adhered to the substrate, and most of the cells were efficiently released. FIG. 7C is a statistical analysis of cell density before and after pH treatment, and it can be seen that the cell release efficiency of the present invention is very high. Meanwhile, the cell viability before and after release is also studied to evaluate the damage degree of the cells during the capturing and releasing process. According to the live/dead staining method, the survival rate of the captured cells is about 98.0%, and the survival rate of the released cells is about 91.5%, which shows that the damage of the captured cells and the released cells to the cells is negligible, and most cells can ensure effective activity, thereby showing the practical value and significance of the invention.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims

1. A novel bionic adhesive material is characterized in that a polypeptide nanofiber containing dopa is self-assembled to form an intelligent adhesive material; the polypeptide nanofiber comprises a self-assembly structure polypeptide sequence and an adhesion group; the self-assembly structure polypeptide sequence forms a nanofiber structure through hydrophobic interaction self-assembly; the adhesion group relies on catechol structure to achieve adhesion.

2. The novel biomimetic adhesive material of claim 1, wherein the self-assembled structural polypeptide sequence is 2-naphthylacetic acid-glycine-phenylalanine (2-NapGFF); the adhesion group is a dopa amino acid; the self-assembly structure polypeptide sequence and the adhesion group form a short peptide containing four amino acids through dehydration condensation.

3. The novel biomimetic adhesive material as claimed in claim 2, wherein the adhesive material is self-assembled in PBS buffer solution to form nanofibers, and further wound and combined to form a fiber network structure.

4. A method for preparing a novel biomimetic adhesive material according to any one of claims 1 to 3, characterized by comprising the following steps:

5. The method for preparing a novel biomimetic adhesive material according to claim 4, wherein step S1 is performed by synthesizing Dopa (TBDMS) by using TBDMS-Cl to protect the structure of DOPAC₂OH, Fmoc-Dopa (TBDMS) satisfying protection of the amino terminus by FmocOSu₂-OH。

6. The method for preparing a novel biomimetic adhesive material according to claim 4, wherein the step S2 is performed in order of adding amino acids including Fmoc-Phe-OH, Fmoc-Phe-OH, and 2-Nap-Gly-OH.

7. Use of a novel biomimetic adhesive material prepared according to the method of claim 4 for high efficiency adhesion as well as non-specific adhesion of biological cells.

8. Use of a novel biomimetic adhesive material prepared according to the method of claim 4 for efficient release of biological cells without damage to the cells.