A combination of lung cancer antigens and its application, cytotoxic T lymphocytes
A lymphocyte and cytotoxic technology, applied in the field of biomedicine, can solve the problems of inaccurate results, separation from experiments, and high incidence of lung cancer
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[0040] 1. Lung cancer-associated neoantigens:
[0041] The whole exome sequencing results of patient Li’s tumor tissue were analyzed for tumor neoantigens (Dingcheng Peptide Source Bio-Information Technology Co., Ltd.), and 80 antigens were synthesized (Nanjing GenScript Polypeptide Synthesis Department);
[0042] Table 1 Peptide sequence and corresponding number
[0043]
[0044]
[0045]
[0046]
[0047] 2. Preparation of APC by co-culture:
[0048] 1) Resuscitated frozen PBMC (2.5×10 7 cells), dilute the freezing solution with 1640+10% FBS, 1500rpm for 5min, resuspend with 10mL1640+10%FBS, 1×10 6 cells / mL, 37°C, 5% CO 2 Recovery culture for 24h;
[0049] 2) Prepare the peptide, 1640+10% FBS+200U / mL IL-2+1600U / mL GM-CSF, the final concentration of the peptide is 50ng / mL, mix well and set aside;
[0050] 3) Collect the cells by centrifugation at 1500rpm for 5min, resuspend the cells with peptide solution, and adjust the density to 1×10 6 cells / mL;
[0051]...
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