Application of DMP nanoparticles in mRNA delivery
A nanoparticle, the use of technology, applied in the field of medicine, to achieve the effect of high transfection rate and low cytotoxicity
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Embodiment 1
[0043] Example 1 Preparation of DMP cationic nanoparticles / IL-22BP mRNA gene complex
[0044] 1. Preparation of DMP nanoparticles by thin film rotary evaporation
[0045] Cationic lipid (2,3-dioleoyl-propyl)-trimethylamine (1,2-dioleoyl-3-trimethylammonium-propane, DOTAP) (structure as figure 1shown) with methyl polyethylene glycol-polycaprolactone (mPEG-PCL) polymer (structure such as figure 2 Shown) (molecular weight is 4000Da, PEG-PCL=2000Da-2000Da) were mixed at a mass ratio of 1:9, and the mixture was dissolved in dichloromethane, and placed on a rotary evaporator at 60°C for 45 minutes film forming. The formed film was taken out and dissolved in deionized water, then shaken in a water bath at 60°C for 5 minutes to obtain a solution of DMP cationic polymer nanoparticles with a specific concentration, which was stored in a 4°C refrigerator for later use.
[0046] 2. Preparation of DMP nanoparticles by microfluidic method
[0047] The cationic lipid (2,3-dioleoyl-propy...
Embodiment 2
[0052] Example 2 Preparation of DMP cationic nanoparticles / Bim mRNA gene complex
[0053] 1. Preparation of DMP nanoparticles by thin film rotary evaporation
[0054] The cationic lipid (2,3-dioleoyl-propyl)-trimethylamine (1,2-dioleoyl-3-trimethylammonium-propane, DOTAP) and methyl polyethylene glycol-polycaprolactone (mPEG-PCL ) high molecular weight polymer (molecular weight is 4000Da, PEG-PCL=2000Da-2000Da) is mixed according to the mass ratio of 1:9, and the mixture is dissolved with dichloromethane, and placed on a rotary evaporator with 60 ℃ rotary evaporation Film formed after 45min. The formed film was taken out and dissolved in deionized water, then shaken in a water bath at 60°C for 5 minutes to obtain a solution of DMP cationic polymer nanoparticles with a specific concentration, which was stored in a 4°C refrigerator for later use.
[0055] 2. Preparation of DMP nanoparticles by microfluidic method
[0056] The cationic lipid (2,3-dioleoyl-propyl)-trimethylamin...
Embodiment 3
[0061] Example 3 Preparation of DMP cationic nanoparticles / IL-15 mRNA gene complex
[0062] 1. Preparation of DMP nanoparticles by thin film rotary evaporation
[0063] The cationic lipid (2,3-dioleoyl-propyl)-trimethylamine (1,2-dioleoyl-3-trimethylammonium-propane, DOTAP) and methyl polyethylene glycol-polycaprolactone (mPEG-PCL ) high molecular weight polymer (molecular weight is 4000Da, PEG-PCL=2000Da-2000Da) is mixed according to the mass ratio of 1:9, and the mixture is dissolved with dichloromethane, and placed on a rotary evaporator with 60 ℃ rotary evaporation Film formed after 45min. The formed film was taken out and dissolved in deionized water, then shaken in a water bath at 60°C for 5 minutes to obtain a solution of DMP cationic polymer nanoparticles with a specific concentration, which was stored in a 4°C refrigerator for later use.
[0064] 2. Preparation of DMP nanoparticles by microfluidic method
[0065] The cationic lipid (2,3-dioleoyl-propyl)-trimethylam...
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