人工碱基核酸适体筛选及其在循环靶标的体外检测中的应用

Aptamer provides a kind of powerful molecular recognition tools for the capture and detection of tumor circulating cells (CTC) and exosomes in biological fluids. However, the aptamers usually suffer from the low screening efficiency, easy biodegradation and general affinity, hindering their applications in the early detection and prognosis of malignant tumors. In order to address the limitations mentioned above, the project will generate a six-base (ATCGZP) nucleic acid library with expanded artificial genetic information systems, resulting in the expansion of library diversity and binding mechanism, and direct an AGEISs cell-SELEX against non-small cell lung cancer to generate multiple biologically stable nucleic acid aptamers with high specificity and high affinity. Furthermore, a microfluidic chip functionalized with artificial base-containing aptamers will be designed and fabricated for highly sensitive and selective detection of CTC and tumor derived exosomes from body fluids. The implementation of the project will provide a panel of new molecular recognition tools and highly sensitive and selective liquid biopsy technology for NSCLC, prompt the applications of aptamer in cancer early detection and prognosis.

核酸适体为生物流体中肿瘤循环细胞（CTC）和外泌体的捕获和检测提供了有力的分子识别工具。然而，核酸适体存在筛选效率低，易生物降解和亲和力一般等局限性，阻碍了其在复杂生物样品检测中的应用。为了解决核酸适体上述局限性，本项目拟通过构建含人工碱基的六碱基（ATCGZP）核酸文库，扩大文库多样性和结合机制，针对非小细胞肺癌细胞发展AGEISs细胞筛选方法，获取多条高特异性、高亲和性和生物稳定好的含人工碱基的核酸适体。进一步将人工碱基核酸适体与微流控芯片技术相结合，对病人血样中循环肿瘤细胞和外泌体进行捕获和检测，以期实现肿瘤细胞的高灵敏、高选择性的快速检测。项目的实施将为非小细胞肺癌的特异性识别提供新的分子工具，为实现肺癌的早期诊断、预后检测提供新的分析手段，促进核酸适体在肿瘤早期诊断和预后中应用。

核酸适体为生物流体中肿瘤循环细胞（CTC）和外泌体的捕获和检测提供了有力的分子识别工具。然而，核酸适体存在筛选效率低，易生物降解和亲和力一般等局限性，阻碍了其在复杂生物样品检测中的应用。为了解决核酸适体上述局限性，本项目通过构建含人工碱基的六碱基（ATCGZP）核酸文库，扩大文库多样性和结合机制，针对非小细胞肺癌细胞发展AGEISs细胞筛选方法，获取了高特异性、高亲和性和生物稳定好的含人工碱基的核酸适体。进一步将人工碱基核酸适体与微流控芯片技术相结合，对病人血样中循环肿瘤细胞和外泌体进行捕获和检测，实现肿瘤细胞的高灵敏、高选择性的快速检测。项目的实施将为非小细胞肺癌的特异性识别提供新的分子工具，为实现肺癌的早期诊断、预后检测提供新的分析手段，促进核酸适体在肿瘤早期诊断和预后中应用。在项目的支持下，课题负责热在SCI源刊物发表4篇学术论文，申请发明专利3项，参与培养博士后和研究生共4人。