Endoglin antibody coupled liposome as well as preparation method and application thereof
An antibody conjugation and liposome technology, which can be used in liposome delivery, preparations for in vivo tests, and medical preparations with inactive ingredients, etc., and can solve problems such as limited application scope
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Embodiment 1
[0055] Embodiment 1: the preparation particle diameter is the plasmid DNA liposome of Endoglin antibody coupling of 100nm, and specific preparation steps are as follows:
[0056] 1) Mix POPC, DDAB, DSPE-PEG with chloroform 2000 and DSPE-PEG 2000 -Maleimide was formulated into corresponding concentrations of 20mg / mL, 5mg / mL, 20mg / mL and 10mg / mL respectively, and mixed in Agilent chromatographic vials in amounts of 18.6μmol, 0.6μmol, 0.6μmol and 0.2μmol; mix well After that, the organic solvent was removed by nitrogen flow for 15 minutes, and the sample bottle was continuously rotated until a thin layer of phospholipids was formed; then placed in a rotary evaporator for vacuum evaporation for 2 hours.
[0057] 2) Take out the sample bottle, add 0.2mL TRIS-HCL buffer solution (50mM, pH 7.0), rotate and oscillate slightly, put it in a water bath sonicator for 3min, fully resuspend the phospholipid mixture, and try to avoid the generation of air bubbles; Add dropwise enhanced gre...
Embodiment 2
[0069] Embodiment 2: Preparation of liposomes loaded with LSS670 dye (available from Kodak) coupled with Endoglin antibody with a particle size of 100nm, the specific preparation steps are as follows:
[0070] 1) Mix POPC, DDAB, DSPE-PEG with chloroform 2000 and DSPE-PEG 2000 -Maleimide was formulated into corresponding concentrations of 20mg / mL, 5mg / mL, 20mg / mL and 10mg / mL respectively, and mixed in Agilent chromatographic vials in amounts of 18.6μmol, 0.6μmol, 0.6μmol and 0.2μmol; mix well After that, the organic solvent was removed by nitrogen flow for 15 minutes, and the sample bottle was continuously rotated until a thin layer of phospholipids was formed; then placed in a rotary evaporator for vacuum evaporation for 2 hours.
[0071] 2) Take out the sample bottle, add 0.2mL TRIS-HCL buffer solution (50mM, pH 7.0), rotate and oscillate slightly, put it in a water bath sonicator for 3min, fully resuspend the phospholipid mixture, and try to avoid the generation of air bubb...
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