CN110028552A - A kind of preparation method of self assembly polypeptide and its hydrogel - Google Patents
A kind of preparation method of self assembly polypeptide and its hydrogel Download PDFInfo
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- CN110028552A CN110028552A CN201910310853.8A CN201910310853A CN110028552A CN 110028552 A CN110028552 A CN 110028552A CN 201910310853 A CN201910310853 A CN 201910310853A CN 110028552 A CN110028552 A CN 110028552A
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- polypeptide
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- self assembly
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
Abstract
The invention discloses the preparation methods of a kind of self assembly polypeptide and its hydrogel, belong to technical field of biological materials.Self assembly polypeptide of the invention, the amino acid sequence of polypeptide are Gly-Ile-Trp-Leu-Gly.The present invention breaches the research ideas and methods of polypeptide hydrogel existing both at home and abroad, and the polypeptide sequence length designed and synthesized is short；The preparation method of polypeptide hydrogel is easy, quick；The good biocompatibility of preparation gained polypeptide hydrogel.The present invention provides flexibility to prepare the chemical design of peptide-based hydrogels, provides simplicity for Peptide systhesis, facilitates the development and utilization of peptide-based hydrogels material.
Description
Technical field
The invention belongs to technical field of biological materials, and in particular to the preparation side of a kind of self assembly polypeptide and its hydrogel
Method.
Background technique
The features such as self assembly polypeptide hydrogel is due to its good biocompatibility, invertibity and degradability, it is extensive
Applied to fields such as organizational project, drug delivery and cell culture.The polypeptide forms that hydrogel can be self-assembled into have amino acid ligand
To type polypeptide, β-hair clip type polypeptide, Fluorenylmethoxycarbonyl (Fmoc) peptide and Amphiphilic peptide, but these energy
The peptide section sequence for being enough self-assembled into hydrogel is longer, the non-degradable terminal groups often containing synthesis.Natural oligopeptides (4-8
Amino acid) self assembly is by less research and concern.
Engineering peptide-based hydrogels are to introduce functional molecular in the segment of some natural polypeptides, have its hydrogel certain
Specific function, but the design of most of engineering peptide-based hydrogels is changed in original self assembly polypeptide system now
It makes, design is restricted, therefore it is very necessary and significant for developing new self assembly polypeptide system.
Summary of the invention
In view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to breach the research of polypeptide hydrogel existing both at home and abroad
Idea and method provides the preparation method of a kind of self assembly polypeptide and its hydrogel.The polypeptide sequence length of the self assembly polypeptide
Short, the preparation method of polypeptide hydrogel is easy, quick, the good biocompatibility of obtained polypeptide hydrogel.
To achieve the above object, using following technical scheme:
A kind of self assembly polypeptide, amino acid sequence are as follows: Gly-Ile-Trp-Leu-Gly.
Above-mentioned self assembly polypeptide is preparing the application in hydrogel.
A kind of polypeptide hydrogel carries out gelation shape by adjusting pH and the concentration of the solution that self assembly polypeptide is formed
At hydrogel.
A kind of preparation method of polypeptide hydrogel, specific steps are as follows: by self assembly polypeptide with the PBS buffer solution of certain pH,
It is configured to certain density polypeptide solution, is vortexed to polypeptide powder end and dissolves, in being stored at room temperature gelation.
The pH range of PBS buffer solution is pH 3 ~ 10 in above-mentioned preparation method.
Preferably, the pH of PBS buffer solution is 7 in above-mentioned preparation method.
The concentration range of the middle polypeptide solution of above-mentioned preparation method is 5.5 ~ 14.69mM.
The middle time of repose of above-mentioned preparation method is 15min.
Of the invention is particularly advantageous in that: the preparation method of a kind of self assembly polypeptide and its hydrogel provided by the invention,
The polypeptide sequence length of its self assembly polypeptide is short, and the preparation method of polypeptide hydrogel is easy, quick；The life of gained polypeptide hydrogel
Object compatibility is good；Flexibility is provided to prepare the chemical design of peptide-based hydrogels, simplicity is provided for Peptide systhesis, facilitates peptide
The development and utilization of based aquagel material.
Detailed description of the invention
Fig. 1 is the plastic phasor under different peptide concentrations, condition of different pH.Sol: polypeptide solution；Gel: polypeptide water-setting
Glue.
Fig. 2 is 14.69mM, and the TEM figure of the polypeptide hydrogel of pH3.0, upper right corner illustration is macroscopical plastic figure.
Fig. 3 is 12.85mM, storage modulus/loss modulus figure of the polypeptide hydrogel of pH7.0.G ': storage modulus；G ' ':
Loss modulus.
Fig. 4 is influence diagram of the polypeptide hydrogel to HepG2 cell activity of 14.69mM, pH7.0.Control: control group；GG:
Experimental group.
Specific embodiment
Embodiment 1
A kind of self assembly polypeptide, amino acid sequence are as follows: Gly-Ile-Trp-Leu-Gly.
The preparation method of the self assembly polypeptide are as follows: by Theoretical Design can self assembly polypeptide, and utilize synthesis in solid state skill
Art carries out Peptide systhesis.
Embodiment 2
A kind of preparation method of polypeptide hydrogel, specific steps are as follows: with different pH(pH3, pH4, pH5, pH6, pH7, pH8, pH9,
PH10 PBS buffer solution), the self assembly for being configured to various concentration (5.5,7.3,9.18,11.02,12.85,14.69 mM) are more
Peptide solution is vortexed to polypeptide powder end and dissolves, and in being stored at room temperature overnight, judges whether solution forms polypeptide water-setting by handstand test tube
Glue, if handstand cuvette sample is not fallen off, as polypeptide hydrogel.
The polypeptide hydrogel that the present embodiment prepares, the plastic phasor under different peptide concentrations, condition of different pH
See Fig. 1.Fig. 1 is the plastic phasor of polypeptide hydrogel, it can be seen from the figure that " v " type, i.e. polypeptide is presented in the Gelation Conditions of polypeptide
When concentration is lower, the pH narrow range of plastic, with the increase of concentration, the pH range of plastic increases.Thus, pH3 ~ 10 compared with
Under wide scope, polypeptide can be self-assembly of hydrogel.
Embodiment 3
A kind of polypeptide hydrogel, preparation method are as follows: weigh self assembly polypeptide powder in 2mL pipe, the PBS that pH3.0 is added is slow
Fliud flushing is configured to the polypeptide solution that concentration is 14.69mM, and vortex to powder sufficiently dissolves, and stands 15min at room temperature and polypeptide is made
Hydrogel takes 10 μ L gel drops on copper mesh, draws extra polypeptide hydrogel after 8min with filter paper, next takes 10 μ L phosphorus
Sodium tungstate solution carries out negative staining, draws extra sodium phosphotungstate solution with filter paper after 8min, is dried at room temperature overnight, in transmission electricity
Its micro-structure diagram is observed on illuminating apparatus.
The microscopic appearance of the polypeptide hydrogel of 14.69mM, pH3.0 that the present embodiment prepares and macroscopical plastic figure are shown in figure
2.Self-assembling polypeptide can be can be visually seen from figure and form the hydrogel with the solid fraction sample of three-dimensional net structure, and can from TEM figure
To observe the presence of nanofiber, this is because pentapeptide pH be 7 when, the amino and carboxylic protons at the end N ' and C ', respectively
Two positive charges and negative electrical charge are taken, carries out being self-assembly of nanofiber by electrostatic interaction, fiber is further wound,
" latching " hydrone, ultimately forms the hydrogel with three-dimensional net structure.
Embodiment 4
A kind of polypeptide hydrogel, preparation method are as follows: weigh the polypeptide powder for being self-assembly of hydrogel in 2mL pipe, be added
The PBS buffer solution of pH7, is configured to the polypeptide solution that concentration is 12.85mM, and vortex to powder is sufficiently dissolved, stood at room temperature
Polypeptide hydrogel is made in 15min.
The storage modulus of the polypeptide hydrogel of 12.85mM manufactured in the present embodiment, pH7.0/loss modulus figure is shown in Fig. 3.Fig. 3
The result shows that its storage modulus is greater than loss modulus, illustrate that sample is in gelling properties, and with shear strain (strain)
Increase, storage modulus is gradually decreased with loss modulus, illustrates that the strain that gel can be born is 0.5%.
Embodiment 5
A kind of polypeptide hydrogel, preparation method are as follows: weigh polypeptide powder that 8 are self-assembly of hydrogel in 2mL pipe, add
The PBS buffer solution for entering pH7, is configured to the polypeptide solution that concentration is 14.69mM, and vortex to powder is sufficiently dissolved, stood at room temperature
Polypeptide hydrogel is made in 15min, takes the polypeptide hydrogel addition culture of 2 μ L to have in 96 orifice plates of HepG2 cell and is cultivated,
20 μ L MTT(5mg/mL are added afterwards for 24 hours), continue to cultivate 4 h.After sucking supernatant, 150 μ L DMSO, microwell plate vibration is added in every hole
Device mixing is swung, light absorption value at 570 nm is measured, control group is the blank pair for replacing the polypeptide hydrogel of 2 μ L with the sterile water of 2 μ L
According to that is, without containing polypeptide hydrogel.
Figure is shown in influence of the polypeptide hydrogel of 14.69mM, pH7.0 that the present embodiment prepares to HepG2 cell activity
4.The result shows that polypeptide hydrogel is to HepG2 cytotoxic side effect, activity can reach 100%, illustrates polypeptide hydrogel Fig. 4
With good biocompatibility.
The foregoing is merely preferable embodiment of the invention, all equivalent changes done according to scope of the present invention patent
With modification, it is all covered by the present invention.
SEQUENCE LISTING
<110>University of Fuzhou
<120>preparation method of a kind of self assembly polypeptide and its hydrogel
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 5
<212> PRT
<213>artificial sequence
<400> 1
Gly Ile Trp Leu Gly
1 5
Claims (8)
1. a kind of self assembly polypeptide, it is characterised in that: the amino acid sequence of the self assembly polypeptide are as follows: Gly-Ile-Trp-Leu-
Gly。
2. a kind of self assembly polypeptide as described in claim 1 is preparing the application in hydrogel.
3. a kind of polypeptide hydrogel, it is characterised in that: by adjust self assembly polypeptide formed polypeptide solution pH and concentration come
The hydrogel for carrying out gelation and being formed.
4. a kind of preparation method of polypeptide hydrogel as claimed in claim 3, which is characterized in that specific steps are as follows:
By self assembly polypeptide with the PBS buffer solution of certain pH, it is configured to certain density polypeptide solution, is vortexed molten to polypeptide powder end
Solution, in being stored at room temperature gelation.
5. a kind of preparation method of polypeptide hydrogel according to claim 4, it is characterised in that: in above-mentioned preparation method
The pH range of PBS buffer solution is pH 3 ~ 10.
6. a kind of preparation method of polypeptide hydrogel according to claim 5, it is characterised in that: in above-mentioned preparation method
The pH of PBS buffer solution is 7.
7. a kind of preparation method of polypeptide hydrogel according to claim 4, it is characterised in that: more in above-mentioned preparation method
The concentration range of peptide solution is 5.5 ~ 14.69mM.
8. a kind of preparation method of polypeptide hydrogel according to claim 4, it is characterised in that: quiet in above-mentioned preparation method
Setting the time is 15min.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113234125A (en) * | 2021-05-10 | 2021-08-10 | 华东理工大学 | Self-assembly polypeptide, polypeptide hydrogel, preparation method and application thereof |
| CN113321709A (en) * | 2021-05-18 | 2021-08-31 | 南方医科大学 | Self-assembled polypeptide, slow-release exosome polypeptide hydrogel and preparation method and application thereof |
| CN113321706A (en) * | 2021-07-15 | 2021-08-31 | 南开大学 | Self-assembly polypeptide derivative and hydrogel prepared from same |
| CN113563421A (en) * | 2021-07-30 | 2021-10-29 | 福州大学 | Preparation method of self-assembled polypeptide reversible hydrogel |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113234125A (en) * | 2021-05-10 | 2021-08-10 | 华东理工大学 | Self-assembly polypeptide, polypeptide hydrogel, preparation method and application thereof |
| CN113234125B (en) * | 2021-05-10 | 2022-12-06 | 华东理工大学 | Self-assembly polypeptide, polypeptide hydrogel, preparation method and application thereof |
| CN113321709A (en) * | 2021-05-18 | 2021-08-31 | 南方医科大学 | Self-assembled polypeptide, slow-release exosome polypeptide hydrogel and preparation method and application thereof |
| CN113321706A (en) * | 2021-07-15 | 2021-08-31 | 南开大学 | Self-assembly polypeptide derivative and hydrogel prepared from same |
| CN113321706B (en) * | 2021-07-15 | 2022-01-07 | 南开大学 | Self-assembly polypeptide derivative and hydrogel prepared from same |
| CN113563421A (en) * | 2021-07-30 | 2021-10-29 | 福州大学 | Preparation method of self-assembled polypeptide reversible hydrogel |
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