Abstract：
Molecularly imprinted polymers (MIPs) are artificial receptors equipped with selective recognition sites for target molecules. One of the most promising-strategies for protein MIPs relies on the exploitation of short surface-exposed protein fragments, termed epitopes, as templates to imprint binding sites in a polymer scaffold for a desired protein. However, the lack of high-resolution structural data of flexible surface-exposed regions challenges the selection of suitable epitopes. Here, we addressed this drawback by developing a polyscopoletin-based MIP that recognizes recombinant proteins via the widely used Strep-tag II affinity peptide. Electrochemistry, surface-sensitive spectroscopy, and molecular dynamics simulations were employed to ensure an utmost control of the Strep-MIP electrosynthesis. The functionality of this novel platform was verified with two Strep-tag labeled enzymes: an O2-tolerant [NiFe]-hydrogenase, and an alkaline phosphatase. The enzymes preserved their biocatalytic activities after multiple utilization confirming the efficiency of Strep-MIP as a general biocompatible platform to confine recombinant proteins for exploitation in biotechnology.
摘要：
分子印迹聚合物(MIP)是配备有针对靶分子的选择性识别位点的人工受体。蛋白质MIP最有前途的策略之一依赖于利用短的表面暴露的蛋白质片段，称为表位，作为模板，以在聚合物支架中压印所需蛋白质的结合位点。然而,缺乏柔性表面暴露区域的高分辨率结构数据对选择合适的表位提出了挑战.这里,我们通过开发一种基于polycotoletin的MIP来解决这一缺点，该MIP通过广泛使用的Strep-tagII亲和肽识别重组蛋白。电化学,表面敏感光谱学，和分子动力学模拟被用来确保Strep-MIP电合成的最大控制。用两种Strep标记标记的酶验证了这种新型平台的功能：耐O2的[NiFe]氢化酶，和碱性磷酸酶.这些酶在多次利用后保留了其生物催化活性，这证实了Strep-MIP作为通用生物相容性平台的效率，可以限制重组蛋白在生物技术中的利用。
