Drug and tumor whole-cell vaccine for treating or preventing tumor, and preparation methods and applications of drug and whole-cell vaccine
A whole-cell vaccine and tumor cell technology, applied in anti-tumor drugs, drug combinations, pharmaceutical formulations, etc.
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[0120] 1. Preparation and method of IL-15 (amplification and purification of hIL-15 plasmid)
[0121] After the recombinant plasmid was confirmed to be correct by restriction enzyme digestion and sequencing, it was prepared in large quantities with the EndofreeTM plasma Giga kit of Qiagen Company. The method refers to the operation manual. The steps are as follows (the amount of the following buffers is suitable for 2.5L bacterial culture):
[0122] 1) Bacterial culture: the plasmid was inoculated in LB medium containing 100 μg / ml Amp, and cultured overnight at 37°C with shaking;
[0123] 2) Harvest bacteria: Centrifuge at 4000rpm / min for 30 minutes at 4°C, discard the supernatant, open the bottle and invert to drain all the supernatant, and weigh;
[0124] 3) Resuspension: Add 125ml Buffer P1 to the bacterial pellet, shake repeatedly, and blow until the bacteria are completely resuspended;
[0125] 4) Lysis: Add 125ml Buffer P2, gently and thoroughly invert the centrifuge ...
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