Nanometer delivery system and preparation method and application thereof
A delivery system and nano-technology, applied in the field of nano-delivery system and its preparation, can solve problems such as inability to combine gene therapy and unclear tumor antigens
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Embodiment 1
[0084] The preparation of embodiment 1 cationic liposome
[0085] (1) Take a certain amount of DOTAP and cholesterol, dissolve it in chloroform respectively, and prepare a storage solution with a concentration of 10 mg / mL;
[0086] (2) mixing DOTAP and cholesterol according to the molar ratio of 1:1;
[0087] (3) nitrogen volatilizes chloroform, and vacuumizes overnight to form a lipid film;
[0088] (4) Hydrate the lipid film with 50°C distilled water and homogenize it by ultrasonic;
[0089] (5) Squeeze the particle size with 200nm and 100nm filter membranes successively to obtain DOTAP / chol cationic liposomes.
Embodiment 2
[0090] The preparation of embodiment 2 guanidinium modified chitosan
[0091] (1) Prepare 1% hydrochloric acid solution, taking 30mL as an example, add 0.3mL concentrated hydrochloric acid to 29.7mL distilled water;
[0092] (2) Weigh 0.1g chitosan (Mr=30000~300000), add 10mL 1% hydrochloric acid solution to dissolve, and prepare 1% chitosan solution;
[0093] (3) In order to improve the reaction efficiency, the molar ratio of chitosan and dicyandiamide was adjusted to 1:2. The molecular weight of chitosan monomer is 161g / mol, so the molar weight of chitosan monomer is n (chitosan monomer)=0.1g / 161(g / mol)=0.000621nmol, then the mole of dicyandiamide The amount should be 0.001242 nmol, m(dicyandiamide) = 104.447 mg. Add dicyandiamide into 1% chitosan solution, stir to promote dissolution.
[0094] (4) After dicyandiamide and chitosan are completely dissolved, the reaction temperature is increased to 100°C, reacted for 3 hours, and cooled to room temperature to obtain the rea...
Embodiment 3
[0095] Purification and physical characterization of embodiment 3 guanidine-modified chitosan
[0096] (1) The reaction product obtained by cooling is placed in a 3500Da dialysis bag. In order to avoid the solubility of chitosan in an aqueous solution and reduce the precipitation, the dialysis is carried out in a weakly acidic environment (1% hydrochloric acid solution), and the dialysis is carried out for 3 hours to remove the non-participating Reactive dicyandiamide.
[0097] (2) Add 2 times the volume of absolute ethanol to the dialyzed solution to precipitate chitosan, and heat and volatilize the ethanol at 65°C to obtain the target reaction product.
[0098] (3) Part of the solid reaction product was taken out for physical characterization by infrared spectroscopy to identify its structure.
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