A kind of exosome targeting lymphoma cells and its preparation method and application
A technology of lymphoma cells and exosomes, which is applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas, etc., can solve the problems of accurate detection and low drug targeting efficiency, and achieve High drug loading rate, prolonged circulation time in vivo, and good biocompatibility
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Embodiment 1
[0043] A CD22-F (ab') carrying doxorubicin 2 A method for preparing an exosome-targeted drug-carrying system of antibody fragments, the method comprising the following steps:
[0044] Step 1: Select 293T cells of the human kidney epithelial cell line as exosome-derived cells, and replace the culture medium of exosome-derived cells with serum-free medium 12-48 hours before extracting exosomes. After culturing for 24-48 hours, collect the cell suspension;
[0045] Step 2: Centrifuge the cell suspension (300g, 10min, 4°C), discard the cells and debris, take the supernatant and centrifuge again (2000g, 10min, 4°C), take the supernatant and centrifuge again (12000g, 30min, 4°C ), the supernatant after the third centrifugation was transferred to a 100kD ultrafiltration tube to balance, and concentrated by centrifugation (4000g, 15min, 4°C). Centrifuge the concentrate at an ultrahigh speed (100,000g, 90min, 4°C), discard the supernatant, resuspend and wash, then centrifuge at the s...
Embodiment 2
[0058] A CD22-F(ab') carrying cyclophosphamide 2 A method for preparing an exosome drug-carrying system of an antibody fragment, the method comprising the following steps:
[0059] Step 1: Select 293T cells of the human kidney epithelial cell line as exosome-derived cells, and replace the culture medium of exosome-derived cells with serum-free medium 12-48 hours before extracting exosomes. After culturing for 24-48 hours, collect the cell suspension;
[0060] Step 2: Centrifuge the cell suspension (300g, 10min, 4°C), discard the cells and debris, take the supernatant and centrifuge again (2000g, 10min, 4°C), take the supernatant and centrifuge again (12000g, 30min, 4°C ), the supernatant after the third centrifugation was transferred to a 100kD ultrafiltration tube to balance, and concentrated by centrifugation (4000g, 15min, 4°C). Centrifuge the concentrate at an ultrahigh speed (100,000g, 90min, 4°C), discard the supernatant, resuspend and wash, then centrifuge at the same...
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