A kind of decoy nucleic acid cationic liposome carrier and preparation method thereof
A technology of cationic liposomes and blank liposomes, applied in the directions of liposome delivery, pharmaceutical formulations, non-active components of polymer compounds, etc., to achieve the effect of no cytotoxicity
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Embodiment 1
[0037] Example 1 Preparation of Decoy nucleic acid cationic liposomes.
[0038] (1) Get DOPE, DOTAP each 3mg, be dissolved in 20ml chloroform and make lipid solution;
[0039] (2) Put the above lipid solution into a round-bottomed flask, rotate and evaporate under reduced pressure in a constant temperature water bath at 20°C, pay attention to the adjustment of the speed and temperature to avoid bubbles, remove the organic solvent, and form a lipid film, take 3ml 4mM pH 7.4 HEPES and add In the above-mentioned round bottom flask, hydrate at 20°C for 30 minutes, and then sonicate for 30 minutes after hydration, and the ultrasonic power is 100W to make a crude liposome solution; pass the crude liposome solution through a 0.4 μm membrane 10 times, and then through a 0.2 μm membrane 10 times Make 2mg / ml liposome solution.
[0040] (3) Take 2 mg of protamine and 1 mg of Decoy nucleic acid and dissolve them in 1 ml of 4mM pH 7.4 HEPES buffer respectively to prepare protamine and Dec...
Embodiment 2
[0043] Example 2 Preparation of Decoy nucleic acid cationic liposomes.
[0044] (1) Get DOPE, DOTAP each 3mg, be dissolved in 20ml chloroform and make lipid solution;
[0045] (2) Put the above lipid solution into a round-bottomed flask, rotate and evaporate under reduced pressure in a constant temperature water bath at 20°C, pay attention to the adjustment of the speed and temperature to avoid bubbles, remove the organic solvent, and form a lipid film, take 3ml 4mM pH 7.4 HEPES and add In the above-mentioned round bottom flask, hydrate at 20°C for 30 minutes, and then sonicate for 30 minutes after hydration, and the ultrasonic power is 100W to make a crude liposome solution; pass the crude liposome solution through a 0.4 μm membrane 10 times, and then through a 0.2 μm membrane 10 times Make 2mg / ml liposome solution.
[0046] (3) Take 2 mg of protamine and 1 mg of Decoy nucleic acid and dissolve them in 1 ml of 4mM pH 7.4 HEPES buffer respectively to prepare protamine and Dec...
Embodiment 3
[0049] Example 3 Preparation of Decoy nucleic acid cationic liposomes.
[0050] (1) Take DOPE 3mg, DOTAP 12mg, dissolve in 20ml chloroform to make lipid solution;
[0051] (2) Put the above lipid solution into a round-bottomed flask, rotate and evaporate under reduced pressure in a constant temperature water bath at 20°C, pay attention to the adjustment of the speed and temperature to avoid bubbles, remove the organic solvent, and form a lipid film, take 3ml 4mM pH 7.4 HEPES and add In the above-mentioned round bottom flask, hydrate at 20°C for 30 minutes, and then sonicate for 30 minutes after hydration, and the ultrasonic power is 100W to make a crude liposome solution; pass the crude liposome solution through a 0.4 μm membrane 10 times, and then through a 0.2 μm membrane 10 times Make 2mg / ml liposome solution.
[0052] (3) Take 2 mg of protamine and 1 mg of Decoy nucleic acid and dissolve them in 1 ml of 4mM pH 7.4 HEPES buffer respectively to prepare protamine and Decoy n...
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