基于多价适配体的循环肿瘤细胞的高效捕获和无损释放研究

项目名称： 基于多价适配体的循环肿瘤细胞的高效捕获和无损释放研究

项目编号： No.21305154

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 数理科学和化学

项目作者： 王金娥

作者单位： 中国科学院苏州纳米技术与纳米仿生研究所

项目金额： 25万元

中文摘要： 循环肿瘤细胞（CTC）的分离和检测是建立癌症分期和预后、设计个性化治疗和疾病监测的关键。由于外周血中CTC数量极少，因此对其的分离和检测十分困难。本项目将多价适配体、三维纳米结构的基底与带有温度控制芯片的微流控芯片技术有机结合起来，发展一种CTC高效捕获和温控无损释放的全新方法。采用两种方法制备多价适配体，一是将适配体修饰到PAMAM树形高分子上，得到一个PAMAM上交联上多个适配体的多价适配体，二是利用滚环扩增技术合成包含多个适配体重复序列的长链结构的多价适配体。多价的适配体亲和分子可以提高其与靶细胞的亲和力，三维基底能够增强细胞与基底的接触频率和持续时间，温度控制芯片可以无损地释放捕获的细胞。以前列腺癌细胞系为模型，系统考察了所构建系统的捕获靶细胞的效果、纯度及温控释放的效果。本项研究将揭示出多价适配体、三维纳米结构基底、微流控芯片用于高效捕获和无损释放CTC的一般规律和内在机理。

中文关键词： 循环肿瘤细胞；三维纳米结构；细胞捕获和释放；适配体；微流控芯片

英文摘要： The isolation and detection of rare circulating tumor cells (CTCs) is critical to cancer diagnosis and prognosis, personalized therapy, and disease monitoring. However, CTC are extremely rare in peripheral blood - as few as one CTC per billion blood cells, making their isolation and detection technically challenging. Here we will develop a multivalent aptamer-mediated, three-dimensional (3D) nanostructured substrate-based microfluidic device that is not only able to efficiently isolate CTC from unprocessed whole blood, but also able to non-destructive, temperature-mediated release and retrievale of cells. The multivalent aptamers will be prepared by two ways, i) the aptamer functional polyamidoamine (PAMAM) dendrimer will be synthesized by conjugating several aptamers to one PAMAM dendrimer molecule, ii) a long aptamer strand comprising repeating aptamer sequences will be synthesized by rolling circle amplification (RCA) technique. The multivalent aptamers will be used to further enhance the affinity of the aptamers with cells. The 3D nanostructured substrate, a silicon-nanopillar (SiNP) array, which allows for increasing cell/substrate contact frequency and duration, will be prepared by a wet chemical etching method. The cell capture substrates will be prepared by N-hydroxysuccinimide (NHS)/maleimide chemistry

英文关键词： circulating tumor cells；3D nanostructure；cell capture and release；aptamer；microfluidic chip