项目名称： 新型适配体功能化电纺丝增强固相微萃取技术及其应用研究

项目编号： No.21465008

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 数理科学和化学

项目作者： 杜甫佑

作者单位： 桂林理工大学

项目金额： 50万元

中文摘要： 固相微萃取技术（SPME）是一项重要的样品前处理技术。本项目拟发展适配体功能化电纺丝增强SPME新技术，研究其超微量、高灵敏分析生物样品中痕量疾病相关蛋白的新方法。主要研究内容有：1)多羧基聚合物的合成及电纺丝法制备SPME纳米纤维丝涂层；2)热休克蛋白90a、肌红蛋白、溶菌酶和凝血酶等痕量疾病相关蛋白所对应的适配体筛选及合成；3)适配体功能化电纺丝增强SPME探针和薄膜的制备及特性研究；4）适配体功能化电纺丝增强SPME-LC-MS联用分析生物样品中痕量蛋白质的新方法；5）建立多组分同时分析的适配体功能化电纺丝增强SPME-LC-MS在线联用新方法，构建新型SPME萃取蛋白质所对应的热力学和动力学数学模型；6）采用构建的新方法分析临床生物样品，检测痕量疾病相关蛋白，发现新的疾病相关蛋白标志物。总体目标是构建适配体功能化电纺丝增强SPME新技术，为研究生物样品中痕量蛋白质组提供新技术平台。

中文关键词： 样品前处理技术；固相微萃取；核酸适配体；电纺丝技术；液相色谱-质谱技术

英文摘要： Solid-phase microextraction (SPME) is a very important, simple, efficient and solventless sample preparation method. The project is proposed to develop a novel aptamer-functionalized electrospun nanofiber-based SPME, and to study the ultra-trace, high sensitive and rapid method for separation and analysis of trace disease-related proteins from biological samples. The main research works are focused on as follows: (1) to synthesize polymers with multiple -COOH groups and then to prepare the corresponding SPME probes and membranes by electrospinning technique; (2) to select and produce the corresponding aptamers for recognition of heat shock protein 90α (Hsp90α), myoglobin, lysozyme and thrombin; (3) to prepare aptamer-functionalized electrospun nanofiber-based SPME probes and membranes, and to study their corresponding specific characteristics; (4) to develop novel aptamer-functionalized electrospun nanofiber-based SPME-LC-MS method for rapid and accurate determination trace Hsp90, myoglobin，thrombin and lysozyme in biological samples; (5) to develop on-line mutiple-aptamer founctionalized SPME-LC-MS method for simultaneous analysis of multiple analytes, and to investigate the corresponding thermodynamic and kinetic mathematical model of the proposed SPME method; (6) to separate and analyze trace disease-related proteins in biological samples by using the proposed methods and to monitor some new disease-related biomarkers. The object of the project is to develop aptamer functionalized electrospun nanofiber based SPME methods for providing novel analytical plateforms to analyze trace proteins in biological samples.

英文关键词： Sample preparation;Solid phase microextraction;Aptamer;Electrospinning technique;Liquid chromatography-mass spectrometry