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本研究应用微酸法快速洗脱人非小细胞性肺癌细胞系A549细胞表面由MHCI类分子提呈的、CD8~+T细胞识别的肿瘤相关多肽.通过系统比较不同条件下的多肽洗脱效率及洗脱后细胞的存活率,我们认为采用pH3.3的微酸缓冲液,洗脱时间为1min,可获得最适洗脱效果;并且还发现Ⅰ类分子提呈的多肽可于洗脱后24h内重新表达,因此同一群肺癌细胞可连续3d用微酸缓冲液反复洗脱,并且细胞活性不受影响,从而提高了多肽的洗脱产率.
In this study, the micro acid method was used to rapidly elute tumor-associated polypeptides recognized by MHC class I molecules on the surface of human non-small cell lung cancer cell line A549. The polypeptide elution efficiency under different conditions was systematically compared. As for the survival rate of the eluted cells, we believe that the use of a pH 3.3 micro acid buffer with an elution time of 1 min gives the best elution efficiency; and that the polypeptides presented by the class I molecules can be eluted after elution. Re-expressed within 24h, so the same group of lung cancer cells can be repeatedly eluted with a slightly acidic buffer for 3 days, and the cell activity is not affected, thereby increasing the elution yield of the polypeptide.