新型多肽化聚合物刷的构建及其检测基质金属蛋白酶的性能研究

项目名称： 新型多肽化聚合物刷的构建及其检测基质金属蛋白酶的性能研究

项目编号： No.21504006

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 丁小康

作者单位： 北京化工大学

项目金额： 21万元

中文摘要： 多肽化聚合物刷不仅能够保留多肽的生物活性，还具有多肽固定量高的优点，是医用高分子科学领域的研究热点。本项目以申请人的前期研究为基础，提出构建多肽固定量可控、抗血清蛋白非特异性吸附、并对基质金属蛋白酶（MMP-2/9）具有高响应性的新型多肽化聚合物刷，使其具有在复杂介质（如血清）中灵敏检测基质金属蛋白酶的特性。首先，拟基于ATRP法在SiO2纳米颗粒表面构建聚甲基丙烯酸钠（P(MA-Na)）聚合物刷，初步探讨荧光标记的多肽在聚合物刷的固定性能及其对MMP-2/9的响应能力。其次，拟通过无规共聚在P(MA-Na)聚合物刷中引入聚乙二醇（PEG）链段以降低蛋白质非特异性吸附、调控多肽的固定量，并研究PEG链段对MMP-2/9酶促反应动力学的影响机制。本项目可有助于进一步理解多肽化聚合物刷与基质金属蛋白酶的相互作用机制，从而为构建新型高灵敏度蛋白酶检测手段提供新的理论基础和科学依据。

中文关键词： 聚合物刷；酶分析；基质金属蛋白酶；多肽；生物传感器

英文摘要： Peptide-functionalized polymer brushes are hot spots in research due to their high peptide load capacity and they are able to maintain the bioactivity of peptides. This research proposal aims to develop novel peptide-functionalized polymer brushes with low non-specific adsorption of serum proteins and sensitive response to matrix metalloproteinase (MMP-2/9). First, the polymer brushes of poly(sodium methacrylate) (P(MA-Na)) would be grafted on silica nanoparticles via surface initiated atom transfer radical polymerization (SI-ATRP). The P(MA-Na) polymer brushes would be further functionalized by immobilizing FITC-labeled peptides as probes for detecting MMP-2/9. Once the peptide is cleaved, the peptide fragments containing FITC is released to the solution and can be subsequently detected by using a fluorescence microscope. Second, this research proposal also aims to study the role of polyethylene glycol (PEG) in the polymer brushes to reduce nonspecific adsorption of serum proteins and to regulate the protease activity. This research would provide new approaches for further understanding the interactions between polymer brushes and matrix metalloproteinase, which would promote the application of polymer brushes in clinical diagnoses of major diseases.

英文关键词： polymer brush;protease assay;MMP;peptide;biosensor