The present invention relates to a method for preparing a personalized cancer vaccine. In particular, according to the present invention, CTC as well as DNA and RNA or ctDNA and ctRNA are separated or enriched to a certain ratio in a sample for the first time； by using a background cell as a contrast, 13-20 types of DNAs having tumor-specific somatic mutations, RNA, or short-chain peptide, i.e., tumor neoantigen, which can cause a change in a protein sequence and can be closely bind to an human HLA type I or II receptory and TCR, and can further activate CD8+T cells or CD4+T helper cells, are separated and verified so as to facilitate early diagnosis of cancers； furthermore, a personalized cancer vaccine is prepared within 4-6 weeks, and is used for stimulating immune response in a cancerous object. According to the present invention, antigens capable of stimulating anti-cancer immunity can be accurately, quickly and efficiently captured under an almost noninvasive condition, so that sequencing time and introduced errors can be reduced. The present invention has a wide application prospect in the field of tumor treatment.本發明涉及一種個人化癌症疫苗的製備方法。具體地，本發明首次採用樣本中分離富集CTC及其DNA和RNA或ctDNA和ctRNA至一定比例，利用背景細胞作為對照，分離和證實13-20種能引起蛋白序列變化並能與人體HLA類型I或II受體及TCR緊密結合，還能活化CD8+T細胞或CD4+T輔助細胞的含腫瘤特異體細胞突變的DNA，RNA或短肽鏈，即腫瘤新生抗原(neoantigen)，有助於癌症的早期診斷；並且，在4-6週內，製成個人化癌症疫苗，及時用於激發患癌對象的免疫反應。本發明能在幾乎非侵入性的情況下，準確、快速、高效地捕獲能夠激發抗癌免疫的抗原，減少了定序時間以及插入的誤差，在腫瘤治療領域具有廣泛的應用前景。
