石墨烯增效适配体和抗体夹心双特异性敏感膜识别机理及其应用于抗生素生物传感检测基础研究

The subject of this project is the basic researches on the molecular recognition mechanism of bispecific membrane prepared by the sandwich model between aptamer and antibody combing with graphene enhancement and its application to antibiotics biosensor detection.The methods using SELEX technology for the designing, synthesizing and screening of aptamers which can recognize antibiotics specifically are studied. The screening rule is explored. The recognition mechanism of the aptamer molecular recognition system is researched and elucidated by spectrum and Nuclear Magnetic Resonance (NMR).The specificities of aptamer and antibody are combined. The bispecific membrane (grapheme/aptamer/antibody) for antibiotic residues sensor detection was prepared by the sandwich model between aptamer and antibody combining with the enhancement of graphene, which makes the membrane obtain bispecificities of aptamer and antibody.The molecular recognition mechanism of this bispecific membrane are illuminated.The application of grapheme/aptamer/antibody bispecific membrane to antibiotics biosensor detection is studied.The highly sensitive biosensor for antibiotic residues detection in foods are developed base on grapheme/aptamer/antibody bispecific membrane. Thie project will provide theoretical proofs for the application of aptamer to food safety detection.

本项目拟将抗生素核酸适配体的特异性与抗体的特异性相结合，对石墨烯增效核酸适配体和抗体夹心模式的双特异性敏感膜识别机理及其应用于抗生素生物传感检测进行基础研究。利用SELEX 技术对与抗生素有特异识别能力的核酸适配体进行设计、合成与筛选研究，揭示抗生素核酸适配体筛选规律。利用现代波谱、核磁共振等方法，阐明核酸适配体与目标抗生素分子识别机理。抗生素特异性核酸适配体和抗体采用夹心模式，结合石墨烯对灵敏度的增效作用，构建既具有适配体特异性又具有抗体特异性的抗生素双特异性敏感膜-石墨烯/核酸适配体/抗体双特异性敏感膜，进一步阐明该双特异性敏感膜分子识别机理，并对该双特异性敏感膜在抗生素残留生物传感高灵敏检测中的应用进行基础研究。建立石墨烯增效基于抗生素核酸适配体和抗体夹心模式双特异性敏感膜的食品中抗生素残留生物传感高灵敏检测方法，为适配体与抗体相结合的食品安全检测新方法提供理论依据，具有重要意义。

抗生素残留高灵敏检测对于保障食品安全和人类健康具有重要的意义。本项目以核酸适配体与目标物抗生素的生物识别作用为基础，通过设计包含有核酸适配体和特定引物的多种功能核酸探针，利用滚环扩增技术以及核酸内切酶/外切酶循环酶切放大技术等等温核酸扩增技术，结合新型纳米团簇材料，构建了基于目标物抗生素介导酶放大的生物传感新原理，开发了多种简单、快速、低成本且具有高灵敏度和高特异性的抗生素生物传感新技术，为抗生素以及小分子检测方法研究提供了新的理论和技术依据，为抗生素残留以及相关的食品安全分析建立了简便实用的新技术平台。新技术与传统的抗生素检测方法比较，具有操作简单、检测时间短、分析成本低的优势，且无需昂贵的仪器和专业的技术人员，有望应用于抗生素残留的实际样品检测。该项目自立项以来已取得一系列研究进展，发表了以国际著名刊物Chemical Communications, Biosensors & Bioelectronics为代表的SCI论文11篇，授权国家发明专利7项，初步完成了项目预期的各项工作目标。