CN108060178A - A kind of magnetic Nano genophore of nucleocapsid and preparation method thereof - Google Patents
A kind of magnetic Nano genophore of nucleocapsid and preparation method thereof Download PDFInfo
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- CN108060178A CN108060178A CN201711478523.7A CN201711478523A CN108060178A CN 108060178 A CN108060178 A CN 108060178A CN 201711478523 A CN201711478523 A CN 201711478523A CN 108060178 A CN108060178 A CN 108060178A
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Abstract
Magnetic Nano genophore the invention discloses a kind of nucleocapsid and preparation method thereof, the carrier is the proton sponge shell for including magnetic Nano kernel and being coated on outside kernel, magnetic Nano kernel and proton sponge shell are fitted close, and the material of magnetic Nano kernel is Fe3O4, the material of proton sponge shell is polyethyleneimine.The magnetic Nano genophore of the present invention has magnetic response characteristic, significantly enhances the directionality of gene transfection, improves the transfection efficiency of gene, and can recycle, and can reduce the cost of gene transfection.
Description
Technical field
The present invention relates to technical field of biological material, the magnetic Nano genophore of more particularly to a kind of nucleocapsid and its preparation
Method.
Background technology
Genophore is a kind of instrument by gene delivery into cell, and main function is that delivery target gene enters host
Cell simultaneously successfully can be replicated and expressed, and be widely used in biological medicine, cell engineering, enzyme engineering, molecular biosciences
Etc. fields.But common genophore is not recyclable, improves the application cost of genetic engineering.Genophore is difficult to back
The main reason for receipts, is that genophore does not have magnetic responsiveness, can not be in situation existing for magnetic field after gene transfection is completed
It is lower by enriching and recovering.Therefore, there is an urgent need to research and develop a kind of genophore with magnetic responsiveness, the recyclable of genophore is improved
Performance reduces the cost of gene transfection.In addition, current nano material prepares mainly water phase coprecipitation method, nano particle
Dispersiveness and particle size distribution wherein is not homogeneous enough, it is difficult to form the carrier material for being suitable for gene transfection.
The content of the invention
It is an object of the invention to overcome shortcoming in the prior art, provide it is a kind of have magnetic responsiveness, it is recyclable,
The magnetic Nano genophore of the nucleocapsid of size uniform.
Another object of the present invention is to provide the preparation method of the magnetic Nano genophore of above-mentioned nucleocapsid.
The purpose of the present invention is achieved through the following technical solutions：
The magnetic Nano genophore of a kind of nucleocapsid, including magnetic Nano kernel and the proton being coated on outside kernel
Sponge shell, magnetic Nano kernel and proton sponge shell are fitted close, and the material of magnetic Nano kernel is Fe3O4, proton sponge
The material of shell is polyethyleneimine.
In the magnetic Nano genophore of above-mentioned nucleocapsid, the surface of polyethyleneimine shell carries strong positive charge
(zeta current potentials>20mV), magnetic Fe3O4Kernel carries strong negative electrical charge (zeta current potentials<- 20mV), it can be obtained by electrostatic self-assembled
Magnetic Nano genophore with nucleocapsid.
The preparation method of the magnetic Nano genophore of above-mentioned nucleocapsid, comprises the following steps：
(1) soluble divalent iron salt and trivalent iron salt are added in reaction vessel, then adds in the co-melting solvent containing low temperature
Aqueous solution and ammonium hydroxide, reacted, be made magnetic Fe3O4Kernel；
(2) by the magnetic Fe3O4Kernel is added in polyethyleneimine in reaction vessel, is then added in co-melting molten containing low temperature
The aqueous solution of agent, is stirred to react, and obtains the magnetic Nano genophore of nucleocapsid.
The co-melting solvent of low temperature refers to the hydrogen bond receptor (such as quaternary ammonium salt) and hydrogen-bond donor (such as acyl by certain mol proportion
The compounds such as amine, carboxylic acid and polyalcohol) two components that are composed or multicomponent eutectic mixture, solidification point it is substantially less than each
The fusing point of a component pure material.The physicochemical properties and ionic liquid of the co-melting solvent of low temperature are closely similar, therefore can be classified as
A kind of novel ion liquid or ionic liquid analog.The present invention can obtain single-size by using the co-melting solvent of low temperature
Nucleocapsid magnetic Nano genophore.Wherein, the solvent that the co-melting solvent of low temperature is preferably made of choline chloride and urea,
The molar ratio of choline chloride and urea is 1:(1.8~2.2)；In the aqueous solution of the co-melting solvent containing low temperature, the co-melting solvent of low temperature
Mass ratio is 1%~99%.
In step (1), it according to the molal weight ratio of ferro element is 1 that the dosage of divalent iron salt and trivalent iron salt, which is,:(1.6~
2.4)；Divalent iron salt is preferably ferrous sulfate, ferrous nitrate, frerrous chloride；Trivalent iron salt is preferably ferric sulfate, ferric nitrate, chlorine
Change iron.
In step (1), the aqueous solution of the co-melting solvent containing low temperature is first added in, adds ammonium hydroxide.
In step (1), reaction temperature is 60~120 DEG C, and the reaction time is 1~6h.
In step (2), the molecular weight of the polyethyleneimine is 25000~75000.
In step (2), temperature is stirred to react as 20~120 DEG C, and the reaction time is 5min~for 24 hours.
The present invention has the following advantages that compared with prior art and effect：
(1) magnetic Nano genophore of the invention has magnetic response characteristic, significantly enhances the direction of gene transfection
Property improves the transfection efficiency of gene.
(2) magnetic Nano genophore of the invention can recycle, and can reduce the cost of gene transfection.
(3) preparation method of the invention is simple, at low cost.
Description of the drawings
Fig. 1 is the structure diagram of the magnetic Nano genophore of nucleocapsid.
Specific embodiment
Further detailed description is done to the present invention with reference to embodiment, but the implementation of the present invention is not limited to this.
Embodiment 1
By frerrous chloride and iron chloride according to ferro element 1:Then 2 molal weight is added in and contained than adding in reaction vessel
(molar ratio of choline chloride and urea is 1 to the co-melting solvent of low temperature:2) aqueous solution (1% mass), ammonium hydroxide react 1h at 60 DEG C,
The magnetic Fe of the nano-gene carrier of nucleocapsid is made3O4Kernel.
By magnetic Fe3O4Kernel is added in polyethyleneimine (molecular weight 25000-75000) in reaction vessel, Ran Houjia
Entering to contain the co-melting solvent of low temperature, (molar ratio of choline chloride and urea is 1:2) aqueous solution (10% mass), is stirred at 30 DEG C
5min is reacted, obtains the magnetic Nano genophore of nucleocapsid.As shown in Figure 1.
Embodiment 2
By frerrous chloride and iron chloride according to ferro element 1:Then 2 molal weight is added in and contained than adding in reaction vessel
(molar ratio of choline chloride and urea is 1 to the co-melting solvent of low temperature:2) aqueous solution (10% mass), ammonium hydroxide, in 120 DEG C of reactions
The magnetic Fe of the magnetic Nano genophore of nucleocapsid is made in 6h3O4Kernel.
By magnetic Fe3O4Kernel is added in polyethyleneimine (molecular weight 25000-75000) in reaction vessel, Ran Houjia
Entering to contain the co-melting solvent of low temperature, (molar ratio of choline chloride and urea is 1:2) aqueous solution (1% mass) stirs anti-at 30 DEG C
5min is answered, obtains the magnetic Nano genophore of nucleocapsid.
Embodiment 3
By ferrous sulfate and chlorination) according to ferro element 1:Then 2 molal weight is added in than adding in reaction vessel containing low
(molar ratio of choline chloride and urea is 1 to the co-melting solvent of temperature:1.8) aqueous solution (1% mass), ammonium hydroxide react 3h at 90 DEG C,
The magnetic Fe of the magnetic Nano genophore of nucleocapsid is made3O4Kernel.
By magnetic Fe3O4Kernel is added in polyethyleneimine (molecular weight 25000-75000) in reaction vessel, Ran Houjia
Entering to contain the co-melting solvent of low temperature, (molar ratio of choline chloride and urea is 1:1.8) aqueous solution (1% mass), is stirred at 30 DEG C
5min is reacted, obtains the magnetic Nano genophore of nucleocapsid.
Embodiment 4
By soluble frerrous chloride and ferric nitrate according to ferro element 1:2 molal weight is than adding in reaction vessel, so
Containing the co-melting solvent of low temperature, (molar ratio of choline chloride and urea is 1 for addition afterwards:2) aqueous solution (99% mass), ammonium hydroxide, 90
DEG C reaction 3h, be made nucleocapsid magnetic Nano genophore magnetic Fe3O4Kernel.
By magnetic Fe3O4Kernel is added in polyethyleneimine (molecular weight 25000-75000) in reaction vessel, Ran Houjia
Entering to contain the co-melting solvent of low temperature, (molar ratio of choline chloride and urea is 1:2) aqueous solution (99% mass), is stirred at 30 DEG C
5min is reacted, obtains the magnetic Nano genophore of nucleocapsid.
Embodiment 5
By frerrous chloride and ferric sulfate according to ferro element 1:Then 2 molal weight is added in and contained than adding in reaction vessel
(molar ratio of choline chloride and urea is 1 to the co-melting solvent of low temperature:2.2) aqueous solution (50% mass), ammonium hydroxide, in 90 DEG C of reactions
The magnetic Fe of the magnetic Nano genophore of nucleocapsid is made in 3h3O4Kernel.
By magnetic Fe3O4Kernel is added in polyethyleneimine (molecular weight 25000-75000) in reaction vessel, Ran Houjia
Entering to contain the co-melting solvent of low temperature, (molar ratio of choline chloride and urea is 1:2.2) aqueous solution (50%) stirs anti-at 30 DEG C
12h is answered, obtains the magnetic Nano genophore of nucleocapsid.
Claims (10)
1. a kind of magnetic Nano genophore of nucleocapsid, it is characterised in that：Including magnetic Nano kernel and it is coated on interior
Proton sponge shell outside core, magnetic Nano kernel and proton sponge shell are fitted close, and the material of magnetic Nano kernel is
Fe3O4, the material of proton sponge shell is polyethyleneimine.
2. the magnetic Nano genophore of nucleocapsid according to claim 1, it is characterised in that：Polyethyleneimine shell
Surface carry strong positive charge (zeta current potentials>20mV), magnetic Fe3O4Kernel carries strong negative electrical charge (zeta current potentials<- 20mV), lead to
It crosses electrostatic self-assembled and obtains the magnetic Nano genophore with nucleocapsid.
A kind of 3. preparation method of the magnetic Nano genophore of the nucleocapsid described in claim 1 or 2, it is characterised in that bag
Include following step：
(1) soluble divalent iron salt and trivalent iron salt are added in reaction vessel, then adds in the water of the co-melting solvent containing low temperature
Solution and ammonium hydroxide, are reacted, and magnetic Fe is made3O4Kernel；
(2) by the magnetic Fe3O4Kernel is added in polyethyleneimine in reaction vessel, then adds in the co-melting solvent containing low temperature
Aqueous solution is stirred to react, and obtains the magnetic Nano genophore of nucleocapsid.
4. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：It is described
The molar ratio of the solvent that the co-melting solvent of low temperature is made of choline chloride and urea, choline chloride and urea is 1:(1.8~
2.2)。
5. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：Step
(1) in, it according to the molal weight ratio of ferro element is 1 that the dosage of divalent iron salt and trivalent iron salt, which is,:(1.6~2.4).
6. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：Divalent
Molysite is ferrous sulfate, ferrous nitrate or frerrous chloride；Trivalent iron salt is ferric sulfate, ferric nitrate or iron chloride.
7. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：Step
(1) in, the aqueous solution of the co-melting solvent containing low temperature is first added in, adds ammonium hydroxide.
8. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：Step
(1) in, reaction temperature is 60~120 DEG C, and the reaction time is 1~6h.
9. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：Step
(2) in, the molecular weight of the polyethyleneimine is 25000~75000.
10. the preparation method of the magnetic Nano genophore of nucleocapsid according to claim 3, it is characterised in that：Step
Suddenly in (2), temperature is stirred to react as 20~120 DEG C, and the reaction time is 5min~for 24 hours.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109781702A (en) * | 2019-01-18 | 2019-05-21 | 中国人民解放军军事科学院军事医学研究院 | A kind of detection method of magnetic microsphere and preparation method thereof and microorganism |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103255175A (en) * | 2013-05-17 | 2013-08-21 | 四川大学 | Magnetic nanometer gene vector system as well as preparation method and application thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103255175A (en) * | 2013-05-17 | 2013-08-21 | 四川大学 | Magnetic nanometer gene vector system as well as preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 刘婷婷: ""磁性壳聚糖微球的工艺研究"", 《中国学位论文全文数据库》 * |
| 张伟等: ""聚乙烯亚胺改性水基磁性液体的吸附特性"", 《纳米技术与精密工程》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109781702A (en) * | 2019-01-18 | 2019-05-21 | 中国人民解放军军事科学院军事医学研究院 | A kind of detection method of magnetic microsphere and preparation method thereof and microorganism |
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