Preparation and application of natural cell membrane derived novel ultrasonic contrast agent
A technology of ultrasound contrast agent and cell membrane, which is applied in the field of biomedicine and can solve the problems of cumbersome preparation process, poor repeatability, and low sensitivity of target molecules and receptors
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Embodiment 1
[0019] Preparation of platelet ultrasound contrast agent: extract a certain amount of peripheral anticoagulated venous blood into a sterile centrifuge tube, record the initial volume (V 0 mL). Platelet-rich plasma was prepared by two-step centrifugation. For the first time, a centrifugal force of 200×g was used for 10 minutes. Carefully draw the upper layer of plasma to another centrifuge tube and discard the lower layer of red blood cells. The second centrifugation adopts a centrifugal force of 1390×g for 15 minutes. The platelet plasma volume needs to be preserved according to the calculation of retaining 1 / 10 of the initial volume. Discard the excess plasma in the upper layer, and blow the platelet sediment in the lower layer to obtain high-concentration platelet plasma.
[0020] The freshly separated platelet plasma was centrifuged at 12000×g for one second to separate platelets and upper plasma. Record the partial volume of plasma (V 1 mL). Then the lower platelet ...
Embodiment 2
[0022] Preparation of erythrocyte ultrasound contrast agent: draw a certain amount of peripheral anticoagulated venous blood into a sterile centrifuge tube, centrifuge the blood at 2500r / min for 5min, discard the plasma and buffy coat, and wash the erythrocytes with isotonic pH=7.4 phosphate The buffer solution was centrifuged at 2500r / min for 5min, washed repeatedly for 3 times, and the supernatant was discarded to obtain red blood cells; the obtained washed red blood cells were mixed with 800mM trehalose solution at a ratio of 1:1.5, and the seaweed The sugar is loaded into the cells, so that the concentration of trehalose in the cells reaches about 60mM, and the erythrocyte concentrate of trehalose is obtained. Red blood cell concentrate and lyophilization buffer (15% PVP + 4% trehalose + 3% glycerol + 5% BSA + 0.05% HSI) were mixed according to the volume ratio of 1:4, then freeze-dried after aliquoting, and carried out with nitrogen Backfill, seal and store to obtain the ...
Embodiment 3
[0024] Preparation of ultrasound contrast agent for prostate cancer cells (PC-3): collect mammalian sample cells suspended in albumin in 1% phosphate buffer at 4° C., and centrifuge the cell suspension to obtain cell pellets. Cells were resuspended in trehalose isotonic solution and co-incubated for 1 h. Add the cell suspension to the container, cool to 4°C and shake the container to spread the cells evenly and freeze at about -70°C for 1 h. Transfer the container to a lyophilizer for lyophilization and inert gas SF 6 Backfilling is carried out, sealed and preserved to obtain the freeze-dried powder of prostate cancer cell ultrasound contrast agent.
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