基于细胞表面特异识别的多色量子点标记小分子抗体提高食管癌外周血循环肿瘤细胞的检测

项目名称： 基于细胞表面特异识别的多色量子点标记小分子抗体提高食管癌外周血循环肿瘤细胞的检测

项目编号： No.31500756

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 乔媛媛

作者单位： 中国人民解放军总医院

项目金额： 20万元

中文摘要： 循环肿瘤细胞（CTC）在恶性肿瘤的诊断及评价预后等方面具有重要的价值，目前已用于转移性乳癌、前列腺癌及结直肠癌的预后评估。食管癌是我国常见恶性肿瘤之一，而食管癌患者外周血CTC的研究尚处在研究阶段，缺乏高特异性和敏感性的检测方法仍是CTCs应用于临床检测的瓶颈。课题组前期以混合的食管癌细胞为靶，从大容量噬菌体抗体库中获得多株与食管癌细胞相关抗原结合的特异性单链抗体。本项目拟通过原核系统表达和纯化抗体后，将这些单链抗体与特定光谱的量子点偶联，建立一种基于表面特异识别的食管癌外周血CTC的高灵敏度和特异性的检测方法，以期应用量子点标记食管癌细胞单链抗体辅助其他的检测方法，提高食管癌外周血中CTC的检出率，从而有效指导食管癌危险分级、早期诊断、辅助治疗、监测肿瘤复发和转移，实现个体化诊疗。

中文关键词： 循环肿瘤细胞；量子点；单链抗体；免疫磁珠

英文摘要： Circulating tumor cell (CTC) has an important value in the diagnosis and evaluation prognosis of malignant tumors and it has been used to assess the prognosis of metastatic breast cancer, prostate cancer and colorectal cancer. Esophageal cancer is one of common malignant tumors in our country, while the CTC of Esophageal squamous cell carcinoma is still in research stage and the lack of high specificity and sensitivity of the detection methods are still a bottleneck into clinical detection. This project on the basis of previous work is to screen single-chain antibodies from a large phage antibody library, using mixed esophageal cancer cells as targets. The proteins associated single-chain antibodies are expressed in prokaryotic system and optimized expression conditions. After purification proteins by affinity chromatography, the biological functions are analyzed. The tumor cells antigens binding to single-chain antibodies will be identified by protein-protein interaction and mass spectrum methods. Eventually an efficient method will be set up combining scFv with quantum dot to specifically recognize CTC in peripheral blood with high sensitivity and specificity. It will be used to enrich CTCs from the peripheral blood of esophageal cancer patients, guiding disease classification and monitoring tumor recurrence and metastasis, to realize individualized diagnosis and treatment.

英文关键词： circulating tumor cell;quantum-dot-labelled;scFv antibody;immune magnetic beads