乳腺癌疫苗中肿瘤抗原的鉴定.pdf

现代食品科技 Modern Food Science and Technology 2013, Vol.29, No.8 乳腺癌疫苗中肿瘤抗原的鉴定 刘安军，方菁菁，马姗婕 （天津科技大学食品工程与生物技术学院，天津300457） 摘要：鉴定乳腺癌疫苗中激活小鼠抗肿瘤免疫反应的肿瘤抗原。利用蛋白质组学方法根据免疫小鼠血清中存在的特异性抗体寻 找肿瘤疫苗所含的肿瘤抗原。结果表明，肿瘤疫苗免疫小鼠的抗体效价大于6400，证明免疫小鼠血清中抗体含量高，且其抗肿瘤体液 免疫得到增强；SDS结合免疫印迹结果证明，软骨多糖作用后的Ca761细胞，与免疫血清在50 kDa左右出现了特异性结合的蛋 白条带；二维电泳结合免疫印迹确定了激活免疫反应的肿瘤抗原为50 kDa酸性蛋白，经质谱技术鉴定，该肿瘤抗原为α-tubulin 。综上 所述，Ca761细胞中的肿瘤抗原α-tubulin经软骨多糖作用后，其结构及表达量发生了变化，该变化的抗原即为肿瘤疫苗中激活小鼠抗 肿瘤免疫反应的抗原，因此肿瘤疫苗免疫成功小鼠可以阻止肿瘤细胞的侵袭；α-tubulin经软骨多糖作用后其免疫原性增强，因此其可 以开发成为以抗原为基础的疫苗。 关键词：猪软骨多糖；乳腺癌疫苗；肿瘤抗原；免疫蛋白质组学 文章篇号：1673-9078(2013)8-1751-1755 Identification of Tumor Antigen in the Breast Cancer Vaccine LIU An-jun, FANG Jing-jing, MA Shan-jie (College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China) Abstract: The tumor antigen in the breast cancer vaccine which activated the anti-tumor immunity response was determined according to the antigen-specific antibodies existent in the serum of the immunized mice by immunoproteomics analysis. The results showed that the antibody titer of immunized mice serum was over 6400 and the high antibody titer suggested that the humoral immunity on anticancer was strengthened. SDSand western-blot analysis showed that the protein extracted from Ca761 cells treated with cartilage polysaccharide was bonded with the immune serum at an estimated molecular mass of 50 kDa . 2Dand western-blot analysis showed that the tumor antigens activated anti-tumor immune response were acidic protein at 50 kDa. By MALDI-TOF-MS, the reactive protein was identified as α-tubulin. In summary, structure and expression of tumor antigen (α-tubulin) in Ca761 cells was modified with the cartilage polysaccharide treatment,