基于卟啉仿生纳米探针的电致化学发光核酸传感器的研究

项目名称： 基于卟啉仿生纳米探针的电致化学发光核酸传感器的研究

项目编号： No.21305067

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： 邓盛元

作者单位： 南京理工大学

项目金额： 25万元

中文摘要： 核酸分析已在分子诊断、基因治疗等领域发挥重要作用，高灵敏核酸检测方法可实现极低浓度DNA/RNA分析，这其中以酶联信号放大技术尤为突出。该技术目前受制于天然酶标的生物活性、低标记率和储备不稳定性。鉴于许多蛋白酶以金属卟啉为辅基，本项目提出了一种融合卟啉有序纳米组装、纳米信号放大和量子点电致化学发光(ECL)技术为一体的核酸分析新策略。该策略通过卟啉与类石墨烯纳米材料的轴向配位作用构建新颖的仿生纳米探针，通过生物偶联而兼具模拟酶催化活性和生物识别特异性；采用肽核酸等新型靶标，耦合探针及其催化产物对ECL辐射的多重猝灭效应和灵活多样的DNA信号放大技术，实现高灵敏核酸分析；并联用多种先进表征手段探索仿生催化机制，揭示组分间协同效应，继而优化参数以获得最佳分析性能。最终将所设计的体系用于表达癌症相关蛋白特定基因序列的分析，为临床诊断奠定基础，并为纳米生物传感提供新颖、高效、便利且普适的酶替代物。

中文关键词： 人工模拟酶；二维纳米材料；催化活性评价；电致发光探针；生物医学分析

英文摘要： DNA assay has already taken a key place in molecular diagnosis, genetic therapy, etc. Ultrasensitive detection method for nucleotide could realize the determination of DNA/RNA at exceptionally low level, of which the enzyme-linked signal amplification protocol prevailed. However, the bioactivity, low tagging rate and unstability of natural enzymes held it back. In view of the metalloporphyrins as cofactor of several protein enzymes, this project proposes a novel strategy for nucleotide assay by integrating the orderly nanoassembly of porphyrins, nanoscale signal amplification and quantum dots (QDs)-based electrochemiluminescence (ECL) technology. The interaction via axial ligation between porphyrins and graphene-like nanomaterials is employed to fabricate the biomimetic nanoprobe, which would possess mimicking catalytic activity and biorecognition specificity by biocoupling reaction. Meanwhile, sequcences of peptide nucleic acid are employed as the capture probes against nonspecific adsorption of tagging molecules. The ultrasensitve QDs-based ECL DNA assays can thus be achieved by coupling multiple quenching effects of the probe and its catalytic products on ECL emission with versatile DNA-based signal amplification techniques. Various spectroscopic and morphological characterizations are applied for stepwise r

英文关键词： Artificial Enzyme Mimics；Two-Dimensional Nanomaterials；Catalytic Activity Assessment；Electrochemiluminescent Probes；Biomedical Assay