CN108546671A - A kind of separation method of cells in biological samples microcapsule bubble - Google Patents
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Abstract
The present invention provides a kind of separation methods of cells in biological samples microcapsule bubble, belong to field of bioanalysis.This separation method includes：After pretreated biological sample is mixed incubation with lipid granule, the compound or fusions of cell microcapsule bubble lipid granule are formed；Cell microcapsule is steeped into the compound of lipid granule again or fusions are detached from biological sample.This method is steeped using lipid granule capture cell microcapsule, existing lipid probe prize law lipid molecular bad dispersibility, easy-to-assemble defect in biological sample solution when detaching cell microcapsule bubble are overcome, the separative efficiency and purity of cell microcapsule bubble are effectively increased.
Description
Technical field
The present invention relates to field of bioanalysis, in particular to a kind of separation side of cells in biological samples microcapsule bubble
Method.
Background technology
It was discovered by researchers that containing the coated microcapsule bubble of multiple film, this kind of microcapsule bubble in many liquid biological samples
It is interior to include nucleic acid materials and the multiple proteins such as mRNA s and microRNA.By taking excretion body as an example, research confirms, excretion
Body can be used as courier, interior to carry each hatching egg by the signal transduction between the approach mediated cell such as blood, urine, lotion, saliva
In vain, the DNA of nucleic acid or even full-length genome, thus the tolerant state that can usually reflect cell health or disease in excretion body,
And to the influence that peripheral cell generates, the relevant information of the diseases such as tumour is reflected to a certain extent.Effectively divide as a result,
It is of great significance from such cell microcapsule bubble.
Currently, the method for detaching such cell microcapsule bubble mainly has：1) supercentrifugation, this method are needed using expensive
Equipment could realize；2) the polymer precipitation method influence purification process its shortcoming is that the foreign protein obtained is excessive；3) it coagulates
Glue exclusion method, its shortcoming is that low separation efficiency, product purity is not high；4) lipid probe prize law, this method are using coupling
The lipid molecular of biotin is contacted with sample, and lipid molecular is inserted into the phospholipid bilayer of microcapsule bubble, then is coupled with Avidin
Magnetic capture biotin-lipid molecular-microcapsule bubble compound.The disadvantages of this method is that lipid molecular is dispersed in aqueous solution
Difference is easily self-assembled into liposome.The modified of this method be by lipid molecular by silylation modification on the glass substrate, disadvantage
It is the limited efficacy of silanization, the finite surface area of substrate of glass；5) positively charged membrane prize law, this method be using it is positively charged again
Raw fibrination pore membrane and positively charged strong anion exchange resin, capture electronegative microcapsule bubble, the disadvantage is that purity and
Separative efficiency is limited.
As the above analysis, the technology of separation cell microcapsule bubble all respectively has limitation in the prior art, cannot be satisfied and grinds
Study carefully needs.
Invention content
The purpose of the present invention is to provide a kind of separation methods of cells in biological samples microcapsule bubble, it is intended to it is micro- to improve cell
The separative efficiency and purity of vesica.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme：
A kind of separation method of cells in biological samples microcapsule bubble comprising：
After pretreated biological sample is mixed incubation with lipid granule, answering for cell microcapsule bubble-lipid granule is formed
Close object or fusions；The compound of cell microcapsule bubble-lipid granule or fusions are detached from biological sample again.
Compared with prior art, beneficial effects of the present invention for example including：
The separation method of this cells in biological samples microcapsule bubble provided by the invention, by being located in advance to biological sample
Reason, remove biological sample in cell, cell fragment and aggregate, obtain include cell microcapsule bubble biological sample supernatant
Liquid；The biological sample supernatant is mixed into incubation with lipid granule again, during incubation, since the surface of lipid granule has
Lipophilic group or surface have positive charge, can be combined by the cell microcapsule bubble in affinity interaction and supernatant or
Person blends with the cell microcapsule bubble in supernatant, forms the compound or fusions of cell microcapsule bubble-lipid granule；Due to this
Kind cell microcapsule bubble-compound of lipid granule or the quality of fusions and molecular size are much larger than cell microcapsule and steep, therefore can
Using more conventional isolation technics, the compound or fusions and separation of the supernatant of cell microcapsule bubble-lipid granule can be made,
Obtain carrying the compound or fusions of cell microcapsule bubble；It can further be cracked, therefrom extract nucleic acid or protein, be used
In a variety of bioanalysis.
This method is steeped using lipid granule capture cell microcapsule, overcomes existing lipid probe prize law in separation cell
Lipid molecular bad dispersibility, easy-to-assemble defect in biological sample solution when microcapsule bubble effectively increases cell microcapsule bubble
Separative efficiency and purity.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described.
Fig. 1 is the egg of the albumen and the extracellular vesica obtained by standard ultracentrifugation method of the extracellular vesica of 1 gained of embodiment
In vain, Western blot analyses, the result of obtained extracellular vesica marker content are carried out at the same time.
Fig. 2 is that the excretion body RNA of 2 gained of embodiment carries out quantitative fluorescent PCR, and obtained β-actin mRNA amplifications are bent
Line.
Fig. 3 is that the excretion body RNA of 2 gained of embodiment carries out quantitative fluorescent PCR, and obtained β-actin mRNA dissolvings are bent
Line.
Fig. 4 is to carry out quantitative fluorescent PCR, obtained β-actin using the excretion body RNA obtained by standard ultracentrifugation method
MRNA amplification curves.
Fig. 5 is to carry out quantitative fluorescent PCR, obtained β-actin using the excretion body RNA obtained by standard ultracentrifugation method
MRNA solubility curves.
Fig. 6 is the excretion body RNA and the excretion body RNA obtained by standard ultracentrifugation method of 2 gained of embodiment, is carried out at the same time glimmering
Fluorescent Quantitative PCR, obtained β-actin mRNA relative amount statistical results.
Specific implementation mode
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Present embodiment provides a kind of separation method of cells in biological samples microcapsule bubble, micro- with the cell for obtaining high-purity
Vesica, and detach the nucleic acid or protein progress routine bioanalytical of its carrying.
Wherein, cell microcapsule bubble is that cell is secreted, falls off or generated after broken, has lipid membrane structure.Cell is micro-
Vesica includes extracellular vesica, particle (100~1000nm), the vesica that falls off (100~1000nm).Wherein, extracellular vesica packet
Include excretion body (40~150nm), microcapsule bubble (100~1000nm), apoptotic body.
Wherein, biological sample include blood plasma, serum, urine, cerebrospinal fluid, saliva, sweat, sperm, tissue hydrops, cell or
Tissue culture supernatant.
The separation method includes the following steps：
Step S1：After pretreated biological sample is mixed incubation with lipid granule, cell microcapsule bubble-lipid is formed
The compound or fusions of grain.
Further, the preprocess method of biological sample includes：Biological sample is filtered or is centrifuged, to remove sample
In cell, cell fragment and aggregate, obtain biological sample supernatant.Preferably, 0.2~0.8 μm of filter is selected when filtering
Film or 0.3~0.7 μm of filter membrane or 0.4~0.6 μm of filter membrane.Preferably, in 1PA18000431SC~16000g, 4~25
10~30min is centrifuged at DEG C, or to centrifuge 15~25min at 12000~14000g, 3~7 DEG C.
Further, in the preferred embodiment, the grain size of lipid granule is 100nm~10 μm, or is the μ of 500nm~8
M is either 1 μm~8 μm or is 3~6 μm.
Further, in some embodiments, the surface modification of lipid granule have cholesterol or steroid derivatives, C6~
At least one of C24 aliphatic acid or its ester, chitosan, peptide, antibody, incomplete antibody, nano antibody and aptamers.Lipid
Its affinity interaction power between cell microcapsule bubble can be enhanced after grain surface modification.Wherein, derived with cholesterol or steroids
Object, the aliphatic acid of C6~C24 (or C10~C20 or C13~C16) or its ester modify lipid granule, can enhance its with it is thin
Lipotropism between born of the same parents' microcapsule bubble；With chitosan-modified lipid granule, it can be enhanced and made with the electrostatic between cell microcapsule steeps
With；Lipid granule is modified with peptide, antibody, incomplete antibody, nano antibody and aptamers, it can be enhanced between cell microcapsule bubble
Intermolecular specific recognition capability, can be used for detaching certain a kind of surface, there is the cell microcapsule of biomarker to steep.
Further, in other embodiments of the present invention, the surface of lipid granule has amphipathic shell, the amphiphilic
Property shell be by with amphipathic lipoid substance modify lipid granule formed.Wherein, it is preferred that amphipathic lipoid substance packet
Include following at least one：(a) phosphatide or glycolipid of C6~C24 or C10~C20 or C13~C16；(b) cholesterol or class
Steroid derivatives；(c) glyceride.
Further, lipid granule is compound for that can steep the liposome combined, solid lipid particles, fat with the cell microcapsule
Composition granule or combinations thereof.This kind of lipid granule can refer to the conventional method in this field to be synthesized, such as liquid-phase precipitation method,
Hydro-thermal reaction method, microemulsion method, sol-gal process, microwave method, ultrasonic method etc..The surface of this kind of lipid granule has lipophilic groups
Group, positive charge can steep the large biological molecule that be specifically bound with cell microcapsule, can by affinity interaction with
Cell microcapsule steeps to form stronger active force, and then during incubation, formed cell microcapsule bubble-lipid granule compound or
Fusions.
Further, it can be following several forms that surface, which has the lipid granule of amphipathic shell,：
A. lipid granule is the liposome of the single-layer or multi-layer imitated vesicle structure formed by amphipathic lipoid substance；
B. the solid lipid particles that lipid granule is made of solid lipid core and amphipathic shell；
Wherein, solid lipid core includes C12~C24 glyceride, the C12~C24 fat for being in solid form at room temperature
At least one of acid and its ester.
C. the fat complexes particle that lipid granule is made of non-lipid core and amphipathic shell；
Wherein, non-lipid core includes by polymer, iron oxide, silica, chitosan, carbon nanotube and Nano carbon balls group
At at least one of group.Preferably, non-lipid core is iron oxide or carbon nanotube.
Using the lipid granule of both forms of B, C, the proportion of core is larger, subsequently by cell microcapsule bubble-lipid
In the compound of particle or the separating step of fusions, centrifugal separation can be used directly, and need not be complex using process
Membrane filtration or air damping method.
Further, biological sample mixed with lipid granule be incubated when temperature be 4~37 DEG C, incubation time 10min
~for 24 hours.It is more preferred, it is incubated using vortex mixer.
Step S2:Again by the compound of cell microcapsule bubble-lipid granule of gained or fusions in step S1 from the life
It is detached in object sample.
Further, it when the compound of cell microcapsule bubble-lipid granule or fusions being detached from biological sample, uses
Membrane filter method, molecular exclusion method or centrifugal process.
Further, when being detached using membrane filter method, use aperture for the filter membrane of 100nm~2 μm.
The feature and performance of the present invention are described in further detail with reference to embodiments：
Embodiment 1
The present embodiment provides a kind of separation methods of extracellular vesica in blood plasma comprising：
A. by blood that EDTA anti-freezings are handled in 1000g centrifugal force, centrifuge 15min at room temperature, remove red in blood
Cell, leucocyte, blood platelet etc., obtain blood plasma.By blood plasma further in the centrifugal force of 1PA18000431SCg, centrifuge at room temperature
30min, removal cell fragment, the vesica being relatively large in diameter and albumen aggregate, obtains supernatant；
B. supersound method is used to synthesize liposome.For example, by N- [1- (2,3- dioleoyl chlorine) propyl]-N, N, N- chlorinations
Trimethylamine (DOTMA) and Distearoyl Phosphatidylethanolamine (DSPE) are according to mass ratio 92:8 are dissolved in the aqueous solution of 4% ethyl alcohol
In, it is placed in ultrasonic emulsification 30min in water-bath.Solvent is removed overnight in merging vacuum drying chamber, and phosphate buffer is added and is made
Positively charged Liposomal suspensions；
C. by 1mL supernatants and 2X1015Positively charged Liposomal suspensions mixing, is placed on rotation vortex mixer, in room temperature
After lower incubation 2h, liposome is combined with extracellular vesica, forms the outer vesicle complexes of liposome-cell or fusions；
D. the polycarbonate membrane for being 100nm with aperture is filtered, and obtains the outer vesicle complexes of liposome-cell or fusion
Object；
E. conventional cracking and extraction agent are used, RNA, DNA contained by extracellular vesica or protein are obtained, is carried out follow-up
Analysis.
F. gained albumen analyzes the content of extracellular vesica marker Alix and TSG101 with Western blot.Meanwhile
Also it uses the supercentrifugation of standard from the 1mL plasma supernatants extracellular vesica of (by pretreatment) extraction and extract proteins, does
Western blot analyses, surpass from obtained supernatant as negative control.The results are shown in Figure 1.It is extracted with this method thin
The content of protein marker is higher than standard ultracentrifugation method in extracellular vesica.
Embodiment 2
The present embodiment provides a kind of separation methods of excretion body in urine comprising：
A. by urine in 2000g centrifugal force, centrifuge 20min at room temperature, remove cell, cell fragment in urine.Again
The membrane filtration for being 0.22 μm with aperture removes the vesica being relatively large in diameter and albumen aggregate therein, obtains filtered fluid；
B. micro emulsion method synthesis of solid lipid granule is used.For example, beaker is put into constant temperature in 70 DEG C of water-baths, sequentially add
80mg glyceryl monooleates, 29mg cetyl palmitates, 20mg ester based quaternary ammonium salts and 1mg DSPE-PEG2000-DBCO,
Magnet rotor stirs, and until whole thawings, is slowly added to 0.5mL hot water.Aforesaid liquid is drawn with syringe, addition is being stirred
4 DEG C of cold water in, obtain solid lipid particles.By 1mgcholesterol-PEG1000-azide and solid lipid particles in phosphorus
It is mixed in phthalate buffer, 4 DEG C of incubation 12h obtain the solid lipid particles of cholesterol modification.
C. by 20mL filtered fluids and 5X1015The solid lipid particles mixing of cholesterol modification, is placed on rotation vortex mixer, in
After being incubated 6h at room temperature, solid lipid particles are combined with excretion body, form solid lipid particles-excretion nanocrystal composition or fusion
Object；
D. the polycarbonate membrane for being 200nm with aperture is filtered, and is obtained solid lipid particles-excretion nanocrystal composition or is melted
Close object；
E. conventional cracking and extraction agent are used, RNA, DNA contained by excretion body or protein is obtained, is subsequently divided
Analysis.
F. gained RNA analyzes the wherein mRNA contents of β-actin with quantitative fluorescent PCR.Meanwhile also using the hypervelocity of standard
Centrifugal process does quantitative fluorescent PCR analysis from 20mL urine filterings liquid (by pretreatment) extraction excretion body and extracting RNA.We
The amplification curve of method and the result of solubility curve are as shown in Figures 2 and 3, the amplification curve of supercentrifugation and the knot of solubility curve
Fruit is as shown in Figure 4 and Figure 5.The mRNA relative amount statistical results of corresponding β-actin are as shown in Figure 6.It is extracted with this method
Excretion body in β-actin mRNA content be higher than standard ultracentrifugation method.
Embodiment 3
The present embodiment provides a kind of separation methods of extracellular vesica in serum comprising：
A. the blood that coagulation vessel acquires is placed in room temperature and waits for that blood solidifies completely, in 1000g centrifugal force, centrifuge at room temperature
10min obtains serum.It is 0.8 μm of membrane filtration, the capsule for removing cell fragment, being relatively large in diameter that serum is further used to aperture
Bubble and albumen aggregate, obtain filtered fluid；
B. freeze-drying synthetic fat composite particles are used.For example, dipalmitoylphosphatidylcholine (DPPC), courage is solid
Alcohol and DSPE-PEG2000-DBCO in molar ratio 200:40:1 mixing, liposome is prepared using supersound method.Liposome is delayed
Slowly it is added drop-wise to the Fe containing 2% Tween-803O4In nano particle suspension, 30min is vibrated, after standing 30min, freeze-drying obtains
Fat complexes particle.By nitrine-Lys-Arg-Gly-Asp small peptide, in molar ratio with fat complexes particle
500:1 mixes in phosphate buffer, and 4 DEG C of incubation 12h obtain short peptide modified fat complexes particle.
C. by 1mL filtered fluids and 1X1016Short peptide modified fat complexes particle mixing, is placed on rotation vortex mixer, in 37
After being incubated 1h at DEG C, fat complexes particle is combined with extracellular vesica, formation fat complexes particle-cell outside vesicle complexes or
Fusions；
D. 5000g centrifuges 10min at room temperature, or uses magnet adsorption, and it is multiple to obtain the outer vesica of fat complexes particle-cell
Close object or fusions；
E. conventional cracking and extraction agent are used, RNA, DNA contained by extracellular vesica or protein are obtained, is carried out follow-up
Analysis.
Embodiment 4
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. it is 0.8 μm of membrane filtration cell culture supernatant to use aperture, removes cell, cell fragment and reunion therein
Object etc., obtains filtered fluid；
B. filtered fluid is mixed with liposome, after being incubated for 24 hours at 25 DEG C, liposome is combined with cell microcapsule bubble, is formed
Liposome-cell microcapsule bubble compound or fusions；
C. the filter membrane for being 100nm with aperture is filtered, and obtains compound or the fusion of cell microcapsule bubble-lipid granule
Object.
Embodiment 5
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. it is 0.2 μm of membrane filtration blood to use aperture, removes cell, cell fragment and aggregate etc. in blood, obtains
To filtered fluid；
B. filtered fluid is mixed with solid lipid particles, after being incubated 10min at 20 DEG C, solid lipid particles are micro- with cell
Vesica combines, and forms solid lipid particles-cell microcapsule bubble compound or fusions；
C. the filter membrane for being 2 μm with aperture is filtered, and obtains compound or the fusion of cell microcapsule bubble-solid lipid particles
Object.
Embodiment 6
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. it is 0.4 μm of membrane filtration biological sample sweat to use aperture, removes cell, cell fragment and reunion in sweat
Object etc., obtains filtered fluid；
B. filtered fluid is mixed with positively charged lipid granule, after being incubated 6h at 23 DEG C, lipid granule is micro- with cell
Vesica combines, and forms lipid granule-cell microcapsule bubble compound or fusions；
C. the filter membrane for being 500nm with aperture is filtered, and obtains compound or the fusion of cell microcapsule bubble-lipid granule
Object.
Embodiment 7
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. by biological sample cerebrospinal fluid in the centrifugal force of 1PA18000431SCg, 30min is centrifuged at 2 DEG C, remove cerebrospinal fluid
In cell, cell fragment and aggregate etc., obtain supernatant；
B. the lipid granule of chitosan mixes with surface modification supernatant, after being incubated 12h at 25 DEG C, lipid granule
It steeps and combines with cell microcapsule, form lipid granule-cell microcapsule bubble compound or fusions；
C. the filter membrane for being 1 μm with aperture is filtered, and obtains the compound or fusions of cell microcapsule bubble-lipid granule.
Embodiment 8
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. by biological sample tissue hydrops in the centrifugal force of 16000g, 10min is centrifuged at 10 DEG C, remove in tissue hydrops
Cell, cell fragment and aggregate etc., obtain supernatant；
B. the lipid granule of glyceride mixes with surface modification supernatant, after being incubated 4h at 24 DEG C, lipid granule
It steeps and combines with cell microcapsule, form lipid granule-cell microcapsule bubble compound or fusions；
C. it is detached with molecular exclusion method, obtains the compound or fusions of cell microcapsule bubble-lipid granule.
Embodiment 9
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. by tissue culture supernatant in the centrifugal force of 12000g, 20min is centrifuged at 4 DEG C, remove in tissue hydrops
Cell, cell fragment and aggregate etc., obtain supernatant；
B. supernatant is mixed with solid lipid particles, after being incubated 2h at 25 DEG C, solid lipid particles and cell microcapsule
Bubble combines, and forms solid lipid particles-cell microcapsule bubble compound or fusions；Wherein, solid lipid particles are by solid fat
The grain structure of matter core and amphipathic shell composition.
C. by the solution obtained by step b in the centrifugal force of 14000g, centrifuge 15min at 4 DEG C, Gu obtain cell microcapsule bubble-
The compound or fusions of body lipid granule.
Embodiment 10
The present embodiment provides a kind of separation methods of cells in biological samples microcapsule bubble comprising：
A. by serum in the centrifugal force of 13000g, 20min is centrifuged at 4 DEG C, remove cell, cell fragment in tissue hydrops
With aggregate etc., supernatant is obtained；
B. supernatant is mixed with fat complexes particle, after being incubated 1h at 23 DEG C, fat complexes particle and cell microcapsule
Bubble combines, and forms fat complexes particle-cell microcapsule bubble compound or fusions；Wherein, fat complexes particle is by non-lipid
The grain structure of core and amphipathic shell composition.
C. by the solution obtained by step b in the centrifugal force of 12000g, centrifuge 25min at 5 DEG C, cell microcapsule bubble-fat is obtained
The compound or fusions of composite particles.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of separation method of cells in biological samples microcapsule bubble, which is characterized in that it includes：
After pretreated biological sample is mixed incubation with lipid granule, the compound of cell microcapsule bubble-lipid granule is formed
Or fusions；The compound of the cell microcapsule bubble-lipid granule or fusions are detached from the biological sample again.
2. the separation method of cells in biological samples microcapsule bubble according to claim 1, which is characterized in that the biology sample
Product and the lipid granule are to mix temperature when being incubated be 4~37 DEG C, incubation time is 10min~for 24 hours.
3. the separation method of cells in biological samples microcapsule bubble according to claim 1, which is characterized in that by the cell
When the compound or fusions of microcapsule bubble-lipid granule are detached from the biological sample, using membrane filter method, molecular exclusion method
Or centrifugal process.
4. the separation method of cells in biological samples microcapsule bubble according to claim 1, which is characterized in that by the cell
When the compound or fusions of microcapsule bubble-lipid granule are detached from the biological sample, use aperture for 100nm~2 μm
Filter membrane is filtered.
5. the separation method of cells in biological samples microcapsule bubble according to claim 1, which is characterized in that the lipid
Grain is that the liposome, solid lipid particles, fat complexes particle or combinations thereof combined can be steeped with the cell microcapsule.
6. the separation method of cells in biological samples microcapsule bubble according to claim 5, which is characterized in that the lipid
The grain size of grain is 100nm~10 μm.
7. the separation method of cells in biological samples microcapsule bubble according to claim 5, which is characterized in that the lipid
Grain surface modification have cholesterol or steroid derivatives, C6~C24 aliphatic acid or its ester, chitosan, peptide, antibody, incomplete antibody,
At least one of nano antibody and aptamers.
8. the separation method of cells in biological samples microcapsule bubble according to claim 5, which is characterized in that the lipid
It is by modifying lipid granule shape with amphipathic lipoid substance that the surface of grain, which has amphipathic shell, the amphipathic shell,
At.
9. the separation method of cells in biological samples microcapsule bubble according to claim 8, which is characterized in that described amphipathic
Lipoid substance includes following at least one：(a) C6~C24 phosphatide or glycolipid；(b) cholesterol or steroid derivatives；(c) sweet
Grease.
10. the separation method of cells in biological samples microcapsule bubble according to claim 1, which is characterized in that the lipid
The core of particle is solid lipid core or non-lipid core, and the solid lipid core includes by C12~C24Glyceride, C12~
C24Aliphatic acid and C12~C24At least one of the group of aliphatic ester composition；The non-lipid core includes by polymer, oxygen
Change at least one of the group of iron, silica, chitosan, carbon nanotube and Nano carbon balls composition.
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Cited By (4)
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| CN109580960A (en) * | 2019-01-14 | 2019-04-05 | 周明 | The separation method of extracellular vesica and the method and kit of the extracellular vesicle surface marker of detection |
| CN113039422A (en) * | 2018-10-17 | 2021-06-25 | 合同会社予幸集团中央研究所 | Method for recovering extracellular vesicles |
| CN113897419A (en) * | 2021-11-08 | 2022-01-07 | 浙江大学 | Kit and method for extracellular vesicle capture or extracellular vesicle quantitative analysis or extracellular vesicle content quantitative analysis |
| RU2803918C1 (en) * | 2023-01-25 | 2023-09-21 | федеральное государственное бюджетное образовательное учреждение высшего образования "Алтайский государственный университет" | Method of producing purified cultured blood serum depleted in growth factors |
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