CN104403101A - Modified polyethyleneimine, preparation method thereof, gene transfection reagent, and applications thereof - Google Patents
Modified polyethyleneimine, preparation method thereof, gene transfection reagent, and applications thereof Download PDFInfo
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- CN104403101A CN104403101A CN201410631399.3A CN201410631399A CN104403101A CN 104403101 A CN104403101 A CN 104403101A CN 201410631399 A CN201410631399 A CN 201410631399A CN 104403101 A CN104403101 A CN 104403101A
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- 229920002873 Polyethylenimine Polymers 0.000 title claims abstract description 84
- 238000012637 gene transfection Methods 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 239000012096 transfection reagent Substances 0.000 title abstract description 6
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 claims abstract description 42
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 11
- 238000007142 ring opening reaction Methods 0.000 claims abstract description 5
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 4
- 125000000467 secondary amino group Chemical group [H]N([*:1])[*:2] 0.000 claims abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 19
- 239000002904 solvent Substances 0.000 claims description 19
- 239000003795 chemical substances by application Substances 0.000 claims description 18
- 239000013612 plasmid Substances 0.000 claims description 17
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical group ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 7
- 108020004707 nucleic acids Proteins 0.000 claims description 7
- 102000039446 nucleic acids Human genes 0.000 claims description 7
- 150000007523 nucleic acids Chemical class 0.000 claims description 7
- 229960001701 chloroform Drugs 0.000 claims description 6
- 238000000746 purification Methods 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- OEOIWYCWCDBOPA-UHFFFAOYSA-N 6-methyl-heptanoic acid Chemical group CC(C)CCCCC(O)=O OEOIWYCWCDBOPA-UHFFFAOYSA-N 0.000 claims description 5
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 claims description 5
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- 238000012986 modification Methods 0.000 claims description 4
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- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 3
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- Medicinal Preparation (AREA)
- Macromolecular Compounds Obtained By Forming Nitrogen-Containing Linkages In General (AREA)
Abstract
The invention discloses modified polyethyleneimine, which comprises polyethyleneimine namely the main chain and lactide that is grafted on the polyethyleneimine. The lactide carries out coordination ring-opening reactions with the primary amine groups or secondary amine groups of the polyethyleneimine, and polymerization reactions do not happen during the process. The invention also discloses a preparation method of the modified polyethyleneimine, a gene transfection reagent, and applications thereof. The experiment results show that the modified polyethyleneimine with the structure mentioned above has a high gene transfection rate, which is greater than that of branched polyethyleneimine on the market under the same conditions, and moreover, no obvious cytotoxicity has been observed. Thus the modified polyethyleneimine can be used as an excellent gene delivery system.
Description
Technical field
The application relates to gene transfection biological technical field, particularly relates to a kind of modified polyethyleneimine and preparation method thereof and gene transfection agent and application thereof.
Background technology
Gene therapy, mainly by exogenous functional gene transfered cell, is modified with this or supplements biological original gene order, changing its protein expression, realize disease treatment object.But exogenous functional gene cannot initiatively be included in by target cell, need to rely on genes delivery system to complete cell and send.Therefore preparing can the carrier system of effective carrying function gene, is the key link of gene therapy.Cationic polymers is as genes delivery system, by electrostatic interactions, the nucleic acid molecule (as plasmid DNA or antisense oligonucleotide) of adsorbs electronegative, forms particle structure and then efficiently carries goal gene, simultaneously by cell endocytic process implementation high-efficiency transfection.
Polymine (PEI) is that the conventional cation radical in laboratory is because of transfection system.It can be divided into straight chain type and branched chain type, and has different molecular weight products.But due to the restriction of PEI self chemical structure, have strong positive polarity, in gene transfection process, easy trigger cell toxicity, causes apoptosis.At present, can transfection efficiency be improved by chemically modified modified PE I further and reduce cytotoxicity.
Summary of the invention
The application provides a kind of modified polyethyleneimine and preparation method thereof and gene transfection agent and application thereof.
According to the first aspect of the application, the application provides a kind of modified polyethyleneimine, comprises the polymine as main chain and the rac-Lactide that is grafted on described polymine; The primary amine of described rac-Lactide and described polymine is amino or secondary amine is amino carries out coordination ring-opening reaction, and non-polymerization reaction take place.
Above-mentioned modified polyethyleneimine, the weight-average molecular weight of described polymine is 600g/mol ~ 75000g/mol.
Above-mentioned modified polyethyleneimine, the structural formula of described modified polyethyleneimine is as follows:
Wherein, x, y, z is nonnegative integer, and x, y and z are all non-vanishing, 14≤x+y≤1744.
Above-mentioned modified polyethyleneimine, the structural formula of described modified polyethyleneimine is as follows:
Wherein, x, y are nonnegative integer, and x and y is all non-vanishing, 14≤x+y≤1744.
According to the second aspect of the application, the application provides a kind of preparation method of modified polyethyleneimine, comprises the steps:
For 1:0.01 ~ 2.7, polymine and rac-Lactide are added in solvent in mass ratio, and add catalyzer and obtain mixture, described mixture is stirred 30 minutes at 25 DEG C, then boiling is heated to, stir 24 hours ~ 72 hours, separation and purification obtains described modified polyethyleneimine, and described modified polyethyleneimine comprises polymine as main chain and the rac-Lactide that is grafted on described polymine; The primary amine amino of described rac-Lactide and described polymine or secondary amine amino coordination open loop grafting, and there is not the ring-opening polymerization of rac-Lactide.
In aforesaid method, described solvent is trichloromethane.
In aforesaid method, the mass ratio of described polymine and described solvent is 1:10 ~ 200.
In aforesaid method, described catalyzer is isocaprylic acid tin.
In aforesaid method, the mass ratio of described polymine and described catalyzer is 1:0.0005 ~ 0.01.
In aforesaid method, the method for described separation and purification is specially: filtered by the reactant of described stirring 24 hours ~ obtain for 72 hours, get filter residue and be dissolved in methanol solution; The methanol solution that will be dissolved with product under room temperature and agitation condition drips anhydrous diethyl ether, centrifugal or filter, collecting precipitation, described in be precipitated as the polymine of rac-Lactide modification.
According to the third aspect of the application, the application provides a kind of gene transfection agent, comprises solvent and is dissolved in the above-mentioned modified polyethyleneimine in described solvent.
Said gene transfection reagent, the mass ratio of described modified polyethyleneimine and solvent is 1 ~ 10:50 ~ 500.
According to the fourth aspect of the application, the application provides a kind of said gene transfection reagent sending the application in thymus nucleic acid, plasmid DNA, Yeast Nucleic Acid or functional protein.
Owing to have employed above technical scheme, the beneficial effect that the application is possessed is:
In the embodiment of the application, with rac-Lactide, chemically modified is carried out to polymine, rac-Lactide at the primary amine of polymine amino or secondary amine amino on coordination open loop obtain modified polyethyleneimine, and there is not the ring-opening polymerization of rac-Lactide.Our experiments show that, the gene transfection rate of the modified polyethyleneimine of this structure is higher, and under equal conditions its gene transfection rate is higher than existing market product branched polyethylene imine, and does not observe obvious cytotoxicity.Therefore, above-mentioned modified polyethyleneimine can be regarded as excellent genes delivery system.
Accompanying drawing explanation
Fig. 1 is the mr collection of illustrative plates of the rac-Lactide modified polyethyleneimine of embodiment 1;
Fig. 2 is the gel retardation assasy detection figure of the mixture that the rac-Lactide modified polyethyleneimine of embodiment 1 and plasmid DNA are formed;
Fig. 3 is that the mixture that formed of the rac-Lactide modified polyethyleneimine of embodiment 1 and Carrying Green Fluorescent Protein plasmid DNA is for HEKC HEK293 cell strain serum-free and the CCK-8 toxicity test detection figure having serum transfection 48h;
Fig. 4 is the gene transfection agent of embodiment 1 rac-Lactide and weight-average molecular weight is that 25000g/mol branched polyethylene imine gene transfection agent carries out Carrying Green Fluorescent Protein plasmid DNA transfection to HEKC HEK293, gene transfection result after 48 hours.
Embodiment
By reference to the accompanying drawings the application is described in further detail below by embodiment.
The modified polyethyleneimine of the application, its a kind of embodiment, comprises the polymine as main chain and the rac-Lactide that is grafted on described polymine; The primary amine of described rac-Lactide and described polymine is amino or secondary amine is amino carries out coordination ring-opening reaction, and non-polymerization reaction take place.In one embodiment, rac-Lactide specifically comprises DL-rac-Lactide and/or L rac-Lactide.
In one embodiment, the weight-average molecular weight of polymine is 600g/mol ~ 75000g/mol.
In one embodiment, the reaction formula generating modified polyethyleneimine is as follows:
Wherein, x, y, z is nonnegative integer, and x, y and z are all non-vanishing, 14≤x+y≤1744.
In another embodiment, the reaction formula generating modified polyethyleneimine is as follows:
Wherein, x, y are nonnegative integer, and x and y is all non-vanishing, 14≤x+y≤1744.
Our experiments show that, the gene transfection rate of the modified polyethyleneimine of this structure is higher, and has less cytotoxicity.Therefore, above-mentioned modified polyethyleneimine can be regarded as excellent genes delivery system.
The preparation method of the modified polyethyleneimine of the application, its a kind of embodiment, comprises the steps:
For 1:0.01 ~ 2.7, polymine and rac-Lactide are added in solvent in mass ratio, and add catalyzer and obtain mixture, described mixture is stirred 30 minutes at 25 DEG C, then boiling is heated to, stir 24 hours ~ 72 hours, separation and purification obtains described modified polyethyleneimine, and described modified polyethyleneimine comprises polymine as main chain and the rac-Lactide that is grafted on described polymine; The primary amine amino of described rac-Lactide and described polymine or secondary amine amino coordination open loop grafting, and there is not the ring-opening polymerization of rac-Lactide.
In one embodiment, solvent is trichloromethane.The mass ratio of polymine and solvent is 1:10 ~ 200.Catalyzer is isocaprylic acid tin.The mass ratio of polymine and catalyzer is 1:0.0005 ~ 0.01.
In one embodiment, the method for separation and purification is specially: filtered by the reactant of stirring 24 hours ~ obtain for 72 hours, get filter residue and be dissolved in methanol solution; Under room temperature and agitation condition, the methanol solution being dissolved with product is dripped anhydrous diethyl ether, centrifugal or filtration, collecting precipitation, is precipitated as the polymine of rac-Lactide modification.
The present invention discloses a kind of preparation method of easy modified polyethyleneimine, realizes rac-Lactide coordination ring-opening reaction on PEI amino, detects do not present assembly effect through nmr spectrometer.Preparation method's productive rate of this modified polyethyleneimine is higher, purifies convenient, is conducive to commercialization and marketing.Under study for action, this modified polyethyleneimine can high-efficiency delivery plasmid DNA, and dye efficiency is far above existing market product polymine (branching, molecular weight 25000g/mol, Sigmaaldrich, CAS 9002-98-6), and do not observe obvious cytotoxicity.
The gene transfection agent of the application, its a kind of embodiment, comprises solvent and is dissolved in the above-mentioned modified polyethyleneimine in solvent.
In one embodiment, the mass ratio of modified polyethyleneimine and solvent is 1 ~ 10:50 ~ 500.
The gene transfection agent of the application is sending the application in thymus nucleic acid, plasmid DNA, Yeast Nucleic Acid or functional protein.
Embodiment 1
1, modified polyethyleneimine is prepared
Be first that the branching PEI (being expressed as bPEI25K) of 25000g/mol is dissolved in the anhydrous trichloromethane of 30 mass parts by the weight-average molecular weight of 1 mass parts, this is mixed in and stirs 30 minutes at 23 DEG C, then 0.01 mass parts rac-Lactide is at room temperature added, finally add 0.0005 mass parts isocaprylic acid tin and obtain mixture, then 76 DEG C are heated to, mixture is seethed with excitement, stirring reaction 72 hours, then filter, the filter residue obtained is the branched polyethylene imine crude product that rac-Lactide is modified, this crude product is dissolved in 4 mass parts Methanol solution, the anhydrous diethyl ether of 50 mass parts is dripped under room temperature and agitation condition, filter, gained throw out is rac-Lactide modified polyethyleneimine, be expressed as bPEI25K-LA.
The structural formula of this modified polyethyleneimine is as follows:
Wherein, x, y, z is nonnegative integer, and x, y and z are all non-vanishing, and x+y+z=581.
Above-mentioned rac-Lactide modified polyethyleneimine (bPEI25K-LA) is detected in deuterated heavy water solvent through magnetic resonance spectrometer.Detected result as shown in Figure 1.Found out by Fig. 1, rac-Lactide does not present assembly effect.
2, gene transfection agent is prepared
The above-mentioned modified polyethyleneimine of 1 mass parts is dissolved in and is mixed with gene in the medical ultrapure water of 50 mass parts and turns reagent.
3, the gel electrophoresis qualification of the complexes carrier (bPEI25K-LA/DNA) of modified polyethyleneimine and DNA plasmid
0.5 micrograms of DNA plasmid (containing Green fluorescent protein fusion vector GFP) solution is added dropwise to a certain amount of said gene to be turned in reagent, with the soft pressure-vaccum mixing of pipettor, room temperature places 30 minutes, make the bPEI25K-LA/DNA complex solution that N/P ratio (N/P) is 0,1,2,5,10,20,40 and 60, after adding 6 × DNA sample-loading buffer, get 10 μ l respectively in 0.8% sepharose after electrophoresis 15 minutes (voltage 120V), observe the retardance situation of DNA in gel, the DNA plasmid that contrast (Control) is equivalent.As shown in Figure 2, N/P is when being greater than 5 in result display electrophoresis result, and band is in loading hole, and illustrates that DNA is arrested in well by the complete inclusion of bPEI25K-LA, confirmation bPEI25K-LA gene transfection agent has higher load efficiency to plasmid DNA.
4, the CCK-8 toxicity test of modified polyethyleneimine
(1) DNA mixture (bPEI25K-LA/DNA) preparation of rac-Lactide modified polyethyleneimine
0.5 micrograms of DNA plasmid (containing GFP reporter gene) solution is added dropwise in appropriate said gene transfection reagent (bPEI25K-LA), with the mixing of pipettor soft pressure-vaccum up and down, room temperature places 30 minutes, make the bPEI25K-LA/DNA complex solution that N/P is 60, normal cell and the cell only adding equal in quality DNA are contrast;
(2) DNA mixture (bPEI25K-LA/DNA) Cytotoxic evaluation of rac-Lactide modified polyethyleneimine
HEKC (HEK293) is with every hole 5 × 10
4the cell density of individual/milliliter is inoculated in 96 orifice plates, by the DMEM culture medium culturing containing 10% foetal calf serum, after cytogamy reaches 80% ~ 90%, every hole adds 100 microlitres respectively and is the Opti-MEM of the bPEI25K/DNA mixture of 60 containing above-mentioned N/P or has serum DMEM culture medium solution, arrange only containing the negative control of DNA plasmid, at 37 DEG C, 5%CO
2cultivate under condition after 6 hours, change liquid with the DMEM substratum containing 10% foetal calf serum, at 37 DEG C, 5%CO
2after continuing to be cultured to 48 hours under condition, by the CCK-8 kit detection cell propagation-toxicity of Dojindo company.Absorbance is surveyed at 450nm place by multi-functional microplate reader.As shown in Figure 3, wherein (a) is serum-free transfection 48 hours to result, and (b) is for there being serum transfection 48 hours figure.
As seen from Figure 3, the toxicity of modified polyethyleneimine to HEKC (HEK293) is less.
5, the in-vitro transfection experiment of modified polyethyleneimine
(1) preparation of bPEI25K-LA/DNA complex solution, bPEI25K/DNA complex solution
0.5 micrograms of DNA plasmid (containing GFP reporter gene) solution is added to appropriate opt i-MEM or has in serum DMEM substratum, final volume is 25 microlitres, leaves standstill 5 minutes; The said gene transfection reagent of same concentrations and bPEI25K gene transfection agent are added to appropriate opt i-MEM or have in serum DMEM substratum, final volume is 25 microlitres, leave standstill after 5 minutes, with pipettor by the mixing of soft for two kinds of solution pressure-vaccum up and down, room temperature places 30 microlitres, make N/P be 60 bPEI25K-LA/DNA mixture and N/P be 60 bPEI25K/DNA mixture.
(2) the outer-gene transfection experiment of modified polyethyleneimine
HEKC (HEK293) is with every hole 5 × 10
4the density of individual/milliliter is inoculated in 96 orifice plates, with containing the DMEM culture medium culturing of 10% foetal calf serum, after cytogamy to 80% ~ 90%, every hole add 50 μ l contain N/P be 60 bPEI25K-LA/DNA mixture, N/P be 60 bPEI25K/DNA mixture, at 37 DEG C, 5%CO
2after hatching 6 hours under condition, change liquid with the DMEM substratum containing 10% foetal calf serum, at 37 DEG C, 5%CO
2cultivation is continued after 48 hours with the green fluorescent protein (GFP) of expressing in inverted fluorescence microscope observation of cell under condition.
As shown in Figure 4: a is that HEK293 cell is having under serum transfection conditions, bPEI25K-LA and the bPEI25K gene transfection agent transfection Carrying Green Fluorescent Protein plasmid DNA experimental result picture of 48 hours.
B be HEK293 cell under serum-free transfection conditions, bPEI25K-LA and the bPEI25K gene transfection agent transfection Carrying Green Fluorescent Protein plasmid DNA experimental result picture of 48 hours.
As seen from Figure 4, bPEI25K-LA gene transfection agent shows good gene transfection performance in HEKC (HEK293).
Can be confirmed by above-mentioned experiment, be that gene transfection agent prepared by Gene transfer vector has the characteristic that toxicity is little, transfection efficiency is high, and application operating is easy by modified polyethyleneimine.
Embodiment 2
Prepare modified polyethyleneimine
First the linear PEI (LPEI0.6K) of the weight-average molecular weight position 600g/mol of 1 weight part is dissolved in the anhydrous trichloromethane of 100 weight part, then 1.5 mass parts rac-Lactides are at room temperature added, after add 0.0025 quality isocaprylic acid tin and obtain mixture, described being mixed in is stirred 25 minutes at 25 DEG C, then 76 DEG C are heated to, mixture is seethed with excitement, stirring reaction 48 hours, filter the branched polyethylene imine crude product of the rac-Lactide modification obtaining being insoluble to chloroform soln, the branched polyethylene imine crude product modified by this rac-Lactide is dissolved in 5 parts by weight Methanol solution, the anhydrous diethyl ether of 120 weight parts is dripped under room temperature and agitation condition, filter, gained throw out is modified polyethyleneimine.
The structural formula of this modified polyethyleneimine is as follows:
Wherein, x, y are nonnegative integer, and x and y is all non-vanishing, and x+y=14.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Above content is the further description done the application in conjunction with concrete embodiment, can not assert that the concrete enforcement of the application is confined to these explanations.For the application person of an ordinary skill in the technical field, under the prerequisite not departing from the application's design, some simple deduction or replace can also be made.
Claims (13)
1. a modified polyethyleneimine, is characterized in that, comprises the polymine as main chain and the rac-Lactide that is grafted on described polymine; The primary amine of described rac-Lactide and described polymine is amino or secondary amine is amino carries out coordination ring-opening reaction, and non-polymerization reaction take place.
2. modified polyethyleneimine as claimed in claim 1, it is characterized in that, the weight-average molecular weight of described polymine is 600g/mol ~ 75000g/mol.
3. modified polyethyleneimine as claimed in claim 1, it is characterized in that, the structural formula of described modified polyethyleneimine is as follows:
Wherein, x, y, z is nonnegative integer, and x, y and z are all non-vanishing, 14≤x+y≤1744.
4. modified polyethyleneimine as claimed in claim 2, it is characterized in that, the structural formula of described modified polyethyleneimine is as follows:
Wherein, x, y are nonnegative integer, and x and y is all non-vanishing, 14≤x+y≤1744.
5. a preparation method for modified polyethyleneimine, comprises the steps:
For 1:0.01 ~ 2.7, polymine and rac-Lactide are added in solvent in mass ratio, and add catalyzer and obtain mixture, described mixture is stirred 30 minutes at 25 DEG C, then boiling is heated to, stir 24 hours ~ 72 hours, separation and purification obtains described modified polyethyleneimine, and described modified polyethyleneimine comprises polymine as main chain and the rac-Lactide that is grafted on described polymine; The primary amine amino of described rac-Lactide and described polymine or secondary amine amino coordination open loop grafting, and there is not the ring-opening polymerization of rac-Lactide.
6. the preparation method of modified polyethyleneimine as claimed in claim 5, it is characterized in that, described solvent is trichloromethane.
7. the preparation method of modified polyethyleneimine as claimed in claim 5, it is characterized in that, the mass ratio of described polymine and described solvent is 1:10 ~ 200.
8. the preparation method of modified polyethyleneimine as claimed in claim 5, it is characterized in that, described catalyzer is isocaprylic acid tin.
9. the preparation method of modified polyethyleneimine as claimed in claim 5, it is characterized in that, the mass ratio of described polymine and described catalyzer is 1:0.0005 ~ 0.01.
10. the preparation method of modified polyethyleneimine as claimed in claim 5, it is characterized in that, the method for described separation and purification is specially: filtered by the reactant of described stirring 24 hours ~ obtain for 72 hours, get filter residue and be dissolved in methanol solution; The methanol solution that will be dissolved with product under room temperature and agitation condition drips anhydrous diethyl ether, centrifugal or filter, collecting precipitation, described in be precipitated as the polymine of rac-Lactide modification.
11. 1 kinds of gene transfection agents, is characterized in that, comprise solvent and are dissolved in the above-mentioned modified polyethyleneimine in described solvent.
12. gene transfection agents as claimed in claim 11, is characterized in that, the mass ratio of described modified polyethyleneimine and solvent is 1 ~ 10:50 ~ 500.
Sending the application in thymus nucleic acid, plasmid DNA, Yeast Nucleic Acid or functional protein as claim 11 or gene transfection agent according to claim 12 for 13. 1 kinds.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410631399.3A CN104403101B (en) | 2014-11-11 | 2014-11-11 | Modified polyethyleneimine and preparation method thereof and gene transfection agent and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410631399.3A CN104403101B (en) | 2014-11-11 | 2014-11-11 | Modified polyethyleneimine and preparation method thereof and gene transfection agent and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104403101A true CN104403101A (en) | 2015-03-11 |
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| US20100270695A1 (en) * | 2006-09-05 | 2010-10-28 | Maciej Radosz | Processing Nanoparticles by Micellization of Blocky-Copolymers in Subcritical and Supercritical Solvents |
| CN103172845A (en) * | 2011-12-23 | 2013-06-26 | 国家纳米科学中心 | Polyethylenimine-aliphatic polyester graft polymer as well as preparation method and nanometer particle thereof |
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| US20100270695A1 (en) * | 2006-09-05 | 2010-10-28 | Maciej Radosz | Processing Nanoparticles by Micellization of Blocky-Copolymers in Subcritical and Supercritical Solvents |
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| CN112759761A (en) * | 2020-12-26 | 2021-05-07 | 广东工业大学 | Modified polyethyleneimine polymer and preparation method thereof |
| CN112759761B (en) * | 2020-12-26 | 2023-02-10 | 广东工业大学 | Modified polyethyleneimine polymer and preparation method thereof |
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