CN109030163A - A kind of fibroin albumen microballoon of wrapping biological sample and preparation method thereof - Google Patents
A kind of fibroin albumen microballoon of wrapping biological sample and preparation method thereof Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- B01J13/02—Making microcapsules or microballoons
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Abstract
The present invention provides the preparation method of the fibroin albumen microballoon of wrapping biological sample, is related to protein microsphere preparation technical field.The preparation method comprises the following steps: the preparation of regenerated silk fibroin solution；Regenerated silk fibroin solution, surfactant, biological sample are mixed, mixed solution is obtained；Mixed solution is sheared after droplet to be scattered in oil-phase solution, by standing the fibroin albumen microballoon that solidifies, be centrifuged, wash, obtain wrapping biological sample after filtration step.The fibroin albumen microballoon no cytotoxicity of wrapping biological sample obtained and it is easy to degrade, realizes encapsulation package, the vital preservation of biological sample.And preparation method is simple, efficient, easily operated, repeatability is strong.
Description
Technical field
The present invention relates to protein microsphere preparation technical fields, and in particular to a kind of fibroin albumen of wrapping biological sample is micro-
Ball and preparation method thereof.
Background technique
With the development of biotechnology, we also go deep into the research of behavior and the regulation of biological sample therewith.It is general and
Speech, cell can only survive under the benign environment for being suitble to its vital movement, and enzyme also can only save its catalysis in suitable environment and live
Property.Therefore, it is necessary to which biological sample to be wrapped up to the adaptive faculty to enhance biological sample to environment, then to the biological sample of package
Product carry out the researchs such as behavior manipulation, active protection, storage transport and functionalization.
Regenerated silk fibroin is derived from a kind of natural polymer of silkworm silk, has very excellent mechanical performance and life
Object compatibility, to cytotoxic side effect, and can in vivo/degrade in vitro.Relative to biomineralization and other height
Molecule coating package, regenerated silk fibroin have its unique biocompatibility and degradability, have in biological sample package
There is very flexible operability, and easily can carry out functionalization arms for different demands, this is in biological sample packet
It wraps up in and there is unrivaled advantage in subsequent applications.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation method of the fibroin albumen microballoon of wrapping biological sample, preparation methods
Simply, efficiently, easily operated, repeatability is strong.
Another object of the present invention is to provide a kind of fibroin albumen microballoon of wrapping biological sample, fibroin albumen microballoon objects
No cytotoxicity and it is easy to degrade, realizes encapsulation package, the vital preservation of biological sample.
Third object of the present invention is the fibroin albumen microballoon for providing a kind of wrapping biological sample in protein function
Application in base group modification and assembling.
The present invention solves its technical problem and adopts the following technical solutions to realize.
The present invention proposes a kind of preparation method of the fibroin albumen microballoon of wrapping biological sample, comprising the following steps:
S1, the preparation of regenerated silk fibroin solution；
The regenerated silk fibroin solution, surfactant, biological sample are mixed, obtain mixed solution by S2, wherein
In the mixed solution, the mass fraction of regenerated silk fibroin is 0.5~5%, and the mass fraction of the surfactant is
0.1-10%；
S3, will the mixed solution shear droplet after to be scattered in oil-phase solution, through standing solidification, centrifugation, washing,
The fibroin albumen microballoon of wrapping biological sample is obtained after filtration step.
The invention proposes a kind of fibroin albumen microballoons of wrapping biological sample, are made according to above-mentioned preparation method.
A kind of beneficial effect of the preparation method and application of the fibroin albumen microballoon of wrapping biological sample of the embodiment of the present invention
Fruit is:
Biological sample is wrapped up using regenerated silk fibroin, is because regenerated silk fibroin has unique biofacies
Capacitive, biological controlled degradation, it is nontoxic nonirritant the advantages that.And biological sample is uniformly encapsulated into regenerated silk fibroin mixing
In the fine droplet of solution, then solidified and stablized in oil-phase solution, ensure that the efficiency and packet of biological sample package
Mild environment during wrapping up in.By the wrapping and encapsulating of fibroin albumen microballoon, delay extraneous thermal conduction rate to a certain extent,
Improve the heat-resistant stability of biological sample.And the biological sample wrapped up is as the limitation in space can not touch direct work
Receptor, biological sample not expression activity under package status, while fibroin albumen microballoon can also completely cut off outer bound pair biological sample
Negative effect, to guarantee that its bioactivity is not destroyed.Moreover, compared to other macromolecule lappings, fibroin egg
Bai Weiqiu is easy to be digested, and the activity of the biological sample after enzymatic hydrolysis can be expressed normally again.Further, fibroin albumen microballoon
Various functional groups can also be wrapped up, functional modification is carried out to fibroin albumen microsphere surface or lining, has fibroin albumen microballoon
There is different functions, to cope with varying environment demand.Can also after by way of layer assembly to solidification fibroin albumen it is micro-
Ball surface carries out the structuring shell assembling of different function.The fibroin albumen method for preparing microsphere of wrapping biological sample of the present invention
Simply, easy to operate, it is low in cost, it can be applied to have very strong in the biomedical articles for preparing different functionalities
Operability is with a wide range of applications.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the flow chart of the preparation method of the fibroin albumen microballoon of wrapping biological sample provided in an embodiment of the present invention；
Fig. 2 is the optical microscopic image of the fibroin albumen microballoon for the package yeast cells that the embodiment of the present invention 1 provides；
Fig. 3 is the fibroin albumen microballoon and Silk fibroin gel, fibroin for the package yeast cells that test example 1 of the present invention provides
The local infrared absorption of protein solution compares map；
Fig. 4 is that the optics under the fibroin albumen microballoon enzymatic hydrolysis state for the package yeast cells that test example 2 of the present invention provides is aobvious
Micro- image；
Fig. 5 is the yeast cell growth curve that the embodiment of the present invention 1 and test example 2 provide.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Below to a kind of preparation method and application of the fibroin albumen microballoon of wrapping biological sample of the embodiment of the present invention into
Row illustrates.
The embodiment of the present invention provides a kind of preparation method of the fibroin albumen microballoon of wrapping biological sample, including following step
It is rapid:
S1, the preparation of regenerated silk fibroin solution；
Further, in the present embodiment, regenerated silk fibroin solution the preparation method comprises the following steps:
S11: putting into after silk is cleaned and carry out degumming in the sodium bicarbonate solution that mass fraction is 0.1~1%, rinsing,
Fibroin fiber is obtained after drying.Preferably, the mass fraction of sodium bicarbonate solution is 0.5%.
S12: the fibroin fiber is placed in the LiBr solution of 9~9.5mol/L, under the conditions of 40~80 DEG C dissolve 2~
6h.Obtain silk fibroin protein solution.Preferentially, fibroin fiber is placed in the LiBr solution of 9.3mol/L, dissolves 4h at 60 DEG C.
S13: the silk fibroin protein solution is placed in the bag filter that molecular cut off is 3000~4000D and is dialysed, is obtained
Dialyzate.Preferably, selecting molecular cut off is the bag filter of 3500D, is dialysed 2~3 days in deionized water.
S14: the dialyzate obtains regenerated silk fibroin solution by separation.Preferably, in regenerated silk fibroin solution
The regenerated silk fibroin for being 5~10% containing mass fraction.
The regenerated silk fibroin solution, surfactant, biological sample are mixed, obtain mixed solution by S2, wherein
In the mixed solution, the mass fraction of regenerated silk fibroin is 0.5~5%, and the mass fraction of the surfactant is
0.1-10%.
Further, in the present embodiment, the surfactant is Pluronick F-127.Pluronick F-127
It is a kind of nonionic surface active agent, it will not influence the activity of biological sample to cell relative non-toxicity, and can make to give birth to
Object sample is evenly dispersed in mixed solution.
Further, in the present embodiment, the biological sample is in cell, bacterium, vaccine, virus, protein
It is one or more.
Further, in the present embodiment, the regeneration for being 5~10% containing mass fraction step S14 obtained
After silk fibroin protein solution is according to requiring to be configured to 0.5~5% regenerated silk fibroin solution, solid Pluronick F- is added
127 are dissolved, and make Pluronick F-127 in the mass fraction 0.1-10% of mixed solution.Different biological samples are pressed
It is added in mixed solution according to different requirement of experiment, biological sample is 5%-50% in the volume accounting of mixed solution.Mixing
Process, which only mildly need to gently shake above-mentioned three kinds of substances, to be shaken up, and this hybrid mode can not only be such that solution is uniformly mixed, also protect
Card will not damage fibroin albumen and biological sample in mixed process.
S3, will the mixed solution shear droplet after to be scattered in oil-phase solution, through standing solidification, centrifugation, washing,
The fibroin albumen microballoon of wrapping biological sample is obtained after filtration step.
Further, shearing droplet is carried out to the mixed solution using micro fluidic device or nozzle-type device.It is preferred that
Ground in the present embodiment, is selected micro fluidic device to carry out droplet shearing to mixed solution, i.e., is added mixed solution and oil-phase solution
Into two pipelines of micro fluidic device right-angled intersection, the flow velocity of two solution of benefit is different, and mixed solution is truncated oil-phase solution
Cut into droplet.Various sizes of droplet can be obtained by adjusting the size of the flow velocity of each pipeline.Preferably, it will mix molten
Liquid cuts into the microballoon that partial size is 500nm-500 μm.
It is understood that in other embodiments, can also will be mixed using the nozzle-type device with rotary nozzle
Liquid rotation is dispersed into the droplet of suitable size during spraying, be then collected in oil-phase solution.
Preferably, the mixed solution of 1 parts by volume is mixed with the oil that the flow velocity of 50-500 μ L/h is distributed to 5~20 parts by volume
In liquid.It is further preferable that the mixed solution is distributed to oil-phase solution, oil-phase solution and mixed solution with the flow velocity of 300 μ L/h
Volume ratio 10:1, flow velocity slow enough guarantees that mixed solution can successfully be cut into droplet and be evenly dispersed in oily phase
In solution, and oil-phase solution is more than mixed solution, and guarantee has enough spaces, and make not will do it to collect between droplet and droplet melts
It closes.
Further, in the present embodiment, according to mass percent meter, the oil-phase solution includes: oleic acid 50%-
90%, organic solvent 5%-30%, poly- ricinoleic acid ester (PGPR) 1%-20%.Preferably, oleic acid 50%, You Jirong
Agent is 30%, PGPR 20%, facilitates to cooperate micro-control stream device to mixed solution according to the oil-phase solution that aforementioned proportion configures
Shearing droplet is carried out, and the organic solvent in oil-phase solution is more conducive to the solidification of fibroin albumen microballoon, improves curing efficiency.
Further, the organic solvent in the oil-phase solution is selected from one of methanol, ethyl alcohol, glycerol and acetone or more
Kind.Preferably, in the present embodiment, the organic solvent in oil-phase solution is ethyl alcohol.
Further, in the present embodiment, mixed solution is dispersed in after oil-phase solution to being placed under the conditions of 4 DEG C and stands
10~15h is solidified, and is then washed, filtration step obtains the fibroin albumen microballoon of wrapping biological sample.
Further, the fibroin albumen microballoon of wrapping biological sample can be placed in water, and be saved under the conditions of 4 DEG C.
The embodiment of the present invention provides a kind of fibroin albumen microballoon of wrapping biological sample, and such as above-mentioned preparation method is made.
It, can be with because fibroin albumen microballoon has good biocompatibility, biological controlled degradation, nontoxic nonirritant
Applied in protein function base group modification and assembling, the fibroin albumen microballoon with different function is prepared.For example, in regenerated silk
Mixed biological sample and ferroferric oxide nano granules carry out cure package in fibroin microballoon, enable to the biology wrapped up
Sample has magnetic target controllable function, all has good enhancing efficiency in targeting control and collection recycling in vitro.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
A kind of fibroin albumen microballoon wrapping up yeast cells provided in this embodiment, obtains according to following steps:
(1) it prepares regenerated silk fibroin solution: silk cocoon is carried out to the bicarbonate of 0.5wt% for putting boiling after removal of impurities processing into
It carries out repeating alkali cleaning twice except glue in sodium solution.Then it rinses 3 times in warm clear water, is dried in 40 DEG C of baking ovens.By drying
Fibroin fiber is placed in the LiBr solution of 9.3mol/L, dissolves 4h in 60 DEG C of baking ovens.Then dissolved fibroin albumen is molten
Liquid is put into the bag filter that interception is 3500D and dialyses 3 days in deionized water, and every 3h changes a water.It is finally obtained by filtration again
Fibroin protein solution saves backup in 4 DEG C of refrigerators.
(2) it configures mixed solution: choosing nature and grow to 107The yeast cells of a/mL concentration is scattered in after being cleaned with PBS
In the solution of regenerated silk fibroin and Pluronick F-127, wherein the mass fraction of regenerated silk fibroin is 5%,
The mass fraction of Pluronick F-127 is 10%.
(3) oil-phase solution: the oleic acid of 50wt%, the ethyl alcohol of 30wt%, the PGPR of 20wt% is configured.
(4) it prepares dispersion liquid: mixed solution and oil-phase solution is transferred in micro fluidic device, adjust mixed solution pipeline
Flow velocity be 300 μ L/h, the droplet that mixed solution cuts into 100 μm is dispersed in oil-phase solution, dispersion liquid is obtained.Its
Middle oil-phase solution and mixed liquor volume ratio are 5:1.
(5) the fibroin albumen microballoon of wrapping biological sample is made: dispersion liquid being placed in 4 DEG C of refrigerators and stands 12h solidification, so
It is rinsed after filtering using ethyl alcohol and deionized water afterwards and obtains the fibroin albumen microballoon of package yeast cells.
(6) it saves: the fibroin albumen microballoon for wrapping up yeast cells being placed in water, is saved under the conditions of 4 DEG C.
Embodiment 2
A kind of fibroin albumen microballoon wrapping up Escherichia coli provided in this embodiment is with the difference place of embodiment 1:
Yeast cells is changed into Escherichia coli in step (2), the mass fraction of regenerated silk fibroin is 2.5%,
The mass fraction of Pluronick F-127 is 5%, and step (4) oil-phase solution and mixed liquor volume ratio are 10:1.
Other steps are same as Example 1 with parameter, obtain the fibroin albumen microballoon of package Escherichia coli.
Can be saved under the conditions of putting in water in 4 DEG C by the Escherichia coli that fibroin albumen microballoon wraps up, with not by
The Escherichia coli of fibroin albumen microballoon package are compared, and the energy of no nutrient environment is resisted by the Escherichia coli that fibroin albumen microballoon wraps up
Power significantly increases, and because there is the protection of fibroin albumen microballoon, the ability for resisting lysozyme is also significantly increased.
Embodiment 3
A kind of package Hela cancer cell/3T3 l cell fibroin albumen microballoon provided in this embodiment, with reality
The difference place for applying example 1 is:
Yeast cells is changed into Hela cancer cell or 3T3 l cell in step (2), and regenerated silk fibroin
The mass fraction that mass fraction is 0.5%, Pluronick F-127 is 0.1%, oil-phase solution and mixed solution in step (4)
Volume ratio is 15:1
Other steps are same as Example 1 with parameter, obtain package Hela cancer cell/3T3 l cell
Fibroin albumen microballoon.
To the package of zooblast, main purpose is based on zooblast to nothing caused by the rigors of living environment
Method saves in normal temperature and pressure, needs to save under condition of ultralow temperature.But the zooblast wrapped up by fibroin albumen microballoon
Can put can save under the conditions of 4 DEG C in water, effectively improve the thermal stability of cell, relax the item of cell storage needs
Part.In addition, having completely cut off the external world by fibroin albumen microballoon package, it is capable of the behavior of regulating cell, and provide one layer of protective cover
It keeps out from the injury such as the physical of the external world, permeability and enzyme.
Embodiment 4
A kind of fibroin albumen microballoon wrapping up JEV japanese encephalitis virus vaccine provided in this embodiment, the area with embodiment 1
Place is not:
Yeast cells is changed into JEV japanese encephalitis virus vaccine in step (2), and oil-phase solution is with mixed liquor volume ratio
20:1
To the carry out fibroin albumen microballoon package of viral vaccine, main purpose is to solve viral vaccine and storing and transporting
Necessary cold chain environment to be offered is spent too high in defeated.By the package of fibroin albumen microballoon, virus or vaccine are under the conditions of 4 DEG C
It can be obtained by, survival ability can significantly improve, and can save the expense of a big chunk cold chain in this way.And release package
The biological effect equally also having the same of vaccine afterwards.
Embodiment 5
A kind of package horseradish peroxidase/ferroferric oxide nano granules provided in this embodiment, the area with embodiment 1
Place is not:
Yeast cells is changed into horseradish peroxidase and/or ferroferric oxide nano granules in step (2).
Carry out fibroin albumen microballoon package to enzyme and protein group, main purpose has: 1. by physical package, is
Biological sample provides one layer of protective case, promotes the environment resistance of these biological samples, and the thermal stability and soda acid such as enzyme are steady
It is qualitative etc.；2. the package by fibroin albumen microballoon integrates different protein groups, making it spatially has association
Same-action, ferroferric oxide nano granules, which are such as added, can enable microballoon have the function of to stab target control, catalase enzyme is added
The hydrogen peroxide etc. being enriched in response environment.
Comparative example 1
This comparative example provides a kind of Silk fibroin gel for wrapping up yeast cells, the preparation method comprises the following steps:
According to the solution that regenerated silk fibroin solution made from 1 step of embodiment (1) and yeast cells are mixed to get, concentration
The Silk fibroin gel for being formed by curing package yeast cells is stood afterwards.
Comparative example 2
This comparative example provides a kind of silk fibroin protein solution for wrapping up yeast cells, the preparation method comprises the following steps:
The solution being mixed to get according to regenerated silk fibroin solution made from 1 step of embodiment (1) and yeast cells.
Test example 1
Test method: the sample that embodiment 1 and comparative example 1,2 are provided carries out infrared absorption pattern analysis.Wherein, silk
Fibroin is in 1580-1720cm-1The infrared absorption pattern of section has different small absorption peaks, represents different fibroin albumen second levels
Structure, wherein just including β-sheets.β-sheets in fibroin albumen secondary structure can characterize fibroin albumen and wrap in solidification
Structure change during wrapping up in, β-sheets content is higher, and structure is more stable, and the ability for resisting the negative effect of external environment is got over
By force.The small absorption peaks different to the fibroin albumen wave band are fitted, and seek area, and different secondary structure occupied area ratios
I.e. different secondary structure proportions.
Fig. 3 is embodiment 1, comparative example 1 and the local infrared absorption comparison diagram of comparative example 2.Fig. 3 (A) is embodiment 1, comparison
Example 1 and comparative example 2 are in 1580-1720cm-1The infrared comparison diagram of section, wherein curve 1 is comparative example 2, and curve 2 is comparative example 1, curve
3 be embodiment 1；Fig. 3 (B) be fibroin albumen include secondary structure in 1580-1720cm-1The distribution diagram of section vibrational band；Fig. 3
(C-E) be comparative example 2 (C), comparative example 1 (D) and embodiment 1 (E) are in 1580-1720cm-1The fitted figure of deconvoluting certainly of section.
Final Fitting Analysis can obtain, the secondary structure β-sheets of fibroin albumen in embodiment 1, comparative example 1 and comparative example 2
Content is respectively 43.85%, 37.23% and 18.49%.As a result it demonstrating, the β-sheets that fibroin albumen microballoon includes is most,
Micro-sphere structure is most stable, it is ensured that the cell wrapped up can more effectively resist the negative effect of external environment.
Test example 2
This test example obtains a kind of fibroin albumen microballoon for wrapping up yeast cells by the step of embodiment 1, carries out following
Test procedure:
(1) the fibroin albumen microballoon addition papain for the package yeast cells that embodiment 1 obtains digest anti-
It answers, obtains the optical microscopic image of the fibroin albumen microballoon of the package yeast cells of different cracking states under Fig. 4 enzymatic hydrolysis condition.Figure
The fibroin albumen microballoon that 4 (A-D) present package yeast cells is cracked since just, cracks half, and half part is cracked to complete
The overall process of cracking.
(2) the fibroin albumen microballoon for wrapping up yeast cells and the yeast cells completely after cracking are placed in YPD culture medium,
It is cultivated at 30 DEG C, cell growth curve measurement then is carried out to yeast cells in above-mentioned two situations, with wavelength 600nm
The absorbance at place characterizes cell solution concentration.
As shown in figure 5, the yeast cells of embodiment 1 obviously maintains an equal level in cell concentration in different time periods.And pass through enzymatic hydrolysis
Afterwards, it is then entered in normal breeding cycle again from the yeast cells discharged in fibroin albumen microballoon, on cell concentration is significant
It rises, and sigmoid curve is presented and meets normal cell proliferation rule.The results show that yeast cells does not express its activity under package status,
Fibroin albumen microballoon can also completely cut off the negative effect of outer bound pair biological sample simultaneously, to guarantee that its bioactivity is not broken
It is bad.And semi-permeable fibroin protein microballoon can allow for the necessary biological micromolecule such as water and oxygen to enter to guarantee it most
Low existence requirement, makes the life yeast cells wrapped up not loss of activity.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of the fibroin albumen microballoon of wrapping biological sample, which comprises the following steps:
S1, the preparation of regenerated silk fibroin solution；
The regenerated silk fibroin solution, surfactant, biological sample are mixed, obtain mixed solution, wherein described by S2
In mixed solution, the mass fraction of regenerated silk fibroin is 0.5~5%, and the mass fraction of the surfactant is 0.1-
10%；
S3 is scattered in oil-phase solution after the mixed solution is sheared droplet, after standing solidification, centrifugation, washing, filtering
Obtain the fibroin albumen microballoon of wrapping biological sample.
2. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that described
The partial size of the fibroin albumen microballoon of wrapping biological sample is 500nm-500 μm.
3. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S2, the biological sample is selected from one of cell, bacterium, vaccine, virus, protein or a variety of.
4. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S2, the surfactant is Pluronick F-127.
5. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S3, the mixed solution is distributed in the oil-phase solution with 50-500 μ L/h flow velocity.
6. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S3, the volume ratio of the oil-phase solution and mixed solution is 5~20:1.
7. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S3, according to mass percent meter, the oil-phase solution includes: oleic acid 50%-90%, organic solvent 5%-30%, poly- castor
Sesame oil acid glyceride 1%-20%.
8. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S3, shearing droplet is carried out to the mixed solution using micro fluidic device or nozzle-type device.
9. the preparation method of the fibroin albumen microballoon of wrapping biological sample according to claim 1, which is characterized in that in step
In rapid S1, the preparation of the regenerated silk fibroin solution the following steps are included:
S11: putting into after silk is cleaned and carry out degumming in the sodium bicarbonate solution that mass fraction is 0.1~1%, rinsing, drying
After obtain fibroin fiber；
S12: the fibroin fiber is placed in the LiBr solution of 9~9.5mol/L, and 2~6h is dissolved under the conditions of 40~80 DEG C,
Obtain silk fibroin protein solution；
S13: the silk fibroin protein solution is placed in the bag filter that molecular cut off is 3000~4000D and is dialysed, is dialysed
Liquid；
S14: the dialyzate obtains regenerated silk fibroin solution by separation.
10. a kind of fibroin albumen microballoon of wrapping biological sample, which is characterized in that such as the preparation of claim 1~9 any one
Method is made.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110368365A (en) * | 2019-08-13 | 2019-10-25 | 西南大学 | A kind of preparation method of the medicine carrying fibroin nanoparticle containing PF127 |
| CN113813893A (en) * | 2021-10-21 | 2021-12-21 | 中国人民解放军军事科学院国防科技创新研究院 | Method for preparing silk fibroin drug delivery capsule based on micro-fluidic chip |
| CN114306737A (en) * | 2021-11-30 | 2022-04-12 | 厦门大学 | Silk fibroin-based porous microsphere and preparation method thereof |
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| CN113813893A (en) * | 2021-10-21 | 2021-12-21 | 中国人民解放军军事科学院国防科技创新研究院 | Method for preparing silk fibroin drug delivery capsule based on micro-fluidic chip |
| CN113813893B (en) * | 2021-10-21 | 2024-04-19 | 中国人民解放军军事科学院国防科技创新研究院 | Method for preparing silk fibroin drug delivery capsule based on microfluidic chip |
| CN114306737A (en) * | 2021-11-30 | 2022-04-12 | 厦门大学 | Silk fibroin-based porous microsphere and preparation method thereof |
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