Ion channel targeted microbubble as well as preparation method and application thereof
An ion channel and targeting technology, applied in the direction of pharmaceutical formulations, preparations for in vivo tests, medical preparations of non-active ingredients, etc., to achieve the effects of easy access, enhanced sensitivity, and improved sensory effects
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preparation example Construction
[0055] The second aspect of the present application provides a method for preparing an ion channel targeting ultrasonic microbubble, including the following steps:
[0056] S01. Preparation of biometric microbubbles;
[0057] S02. Add biotinylated microbubbles to incubate, add biotin-labeled ion channel antibodies, incubation treatment, to obtain ultrasonic microbubbles to ion channels.
[0058] The new type of ionic channel provided by the present application is targeted by the preparation method of ultrasonic microbubbles, first preparing biotinized microbubbles, and then introduces affinity on the surface of biotinylated micrububbles, constructing biotin-affinity system The ion channel targeting ultrasonic microbubbles are prepared by biotin-affinity binding ion channel antibodies. The method is simple to operate, and the ion channel targets ultrasonic micropubbubs are easily obtained, more importantly, the resulting ion channel targets ultrasonic microbubbles, and can target t...
Embodiment 1
[0085] Preparation method of an ion channel targeting ultrasonic microbubble includes:
[0086] (1) 20 mg / ml of diaphragust phosphate choline (DSPC), 20 mg / ml of phosphate ester poly - ethylene glycol 2000 (DSPE-PEG2000) and 20 mg / ml of biological Plasmalated polyethylene glycol 2000 modified diaphragmy ester chloride (DSPE-PEG2000-Biotin) mixed in trichloromethane and mixed on the scroll mixer.
[0087] (2) The organic solvent was removed using a dry nitrogen to form a uniform film on the test tube wall, dried in a vacuum oven for 2-6 hours; then a volume-degassed Tris buffer solution was added to give a phospholipid solution.
[0088] (3) Heating the phospholipid solution to its phase transition temperature (55-60 ° C), and prepared a phospholipid solution with a water bath, dispensed into the small Western whipnee, and the air in the bottle is converted to perfluoropropane, and the mechanical oscillator is shocked. 45S gain biotinylated ultrasonic microbubbles.
[0089] (...
Embodiment 2
[0094] Piezo1 ion channel targeted microbubble sensitive ultrasound 2 Stimulating effect of A cells:
[0095] (1) Providing a Piezo1 ion channel targeting ultrasonic microbubble prepared by the above Example 1;
[0096] (2) Cultivate N 2 A cells are inoculated with 6-well plates, and when they are 75-85% cell density, use CA 2+ Fluorescent probe Fluo 4-AM labeled cells.
[0097] (3) Targeting the PIEZO1 ion channel of step (1) to incubate the cells of the ultrasonic microbubbles and the cells of step (2).
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