利用原位点击化学构建蛋白质印迹识别界面

Molecular imprinting technology is important in preparing functional materials with the ability for molecular recognition. Successful imprinting of proteins would be of great significance in several areas, including bioseparation, biosensing and biomedical engineering. To date, protein imprinting being lack of success is primarily due to harsh reaction system that would alter template conformation prior to polymerization. The current project proposes a click chemistry-based imprinting strategy, by which surface-initiated functional polymer brushes are employed as structural basis. When the polymer brushes go through template-induced assembly in the presence of a target protein, the reaction of in situ crosslinking is carried out to fix their spatial structure. This strategy takes advantage of the moderate conditions of click chemistry-based crosslinking reaction that has proven less destructive for the conformation of proteins. This project aims to establish protein imprinting technique based on the above conception, and clarify the roles of polymer brushes and cross linkers in controlling the structure of imprinting recognition interface. With the established procedure, proteins of large molecule mass, such as human immunoglobulin, are planned to be employed as template for imprinting. This work is anticipated to provide a general solution for protein imprinting.

分子印迹是人工合成具有分子识别能力功能高分子材料的重要途径。成功实现蛋白质分子印迹将会对生物分离、分析传感、生物医学等多个领域产生深远影响。印迹前蛋白质模板结构变形，导致印迹产物选择性不够理想是目前该领域面临的主要问题。本研究提出功能化分子刷在模板蛋白诱导下通过点击化学原位交联成形的印迹策略。其潜在的优势在于点击化学能够提供比自由基聚合更加温和的水相反应环境，而分子刷预聚物的存在可以显著降低印迹反应物浓度，从而克服环境诱导的模板结构变化，提高印迹识别位点的保真度。本项目将基于分子刷预聚物原位点击交联的策略，建立蛋白质分子印迹设计原理和技术体系；阐明分子刷和交联剂对印迹聚合物识别界面的结构调节机制；实现对人免疫球蛋白等大分子量蛋白质的有效印迹。本项目的成功实施有望将基于原位点击化学的多元交联方法扩充到印迹化学的工具箱中，为大分子量蛋白质印迹提供了一个新的思路。

本项目针对蛋白质分子印迹过程中蛋白质结构不稳定的关键问题，探索了一种条件温和的印迹策略。按照项目既定目标，主要完成了以下三部分研究内容：（1）建立了分子刷预聚物修饰微纳米颗粒作为蛋白质印迹载体的技术方法；（2）针对不同模型蛋白，完成了吸附功能团的筛选、固载和吸附性能评价工作，重点研究了疏水功能基的超分子置换洗脱策略，从而使疏水基团能够被纳入到分子印迹吸附功能团的选择范围；（3）探索了两种分子刷聚合物点击交联的技术方法，其中利用炔基功能化水溶性大分子交联叠氮化分子刷的策略被证明是一种具有潜力的蛋白质表面印迹策略。