Fluorogenic monomer activation for protein-initiated atom transfer radical polymerization
Fluorogenic monomer activation for protein-initiated atom transfer radical polymerization
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DOI：
10.1039/d2ob00175f
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发表时间：
2022-06-08
影响因子：
3.2
通讯作者：
Cooley，Christina B.
中科院分区：
文献类型：
--
作者：
Tahseen，Danyal;Sackey-Addo，Jemima R.;Cooley，Christina B.
荧光原子转移自由基聚合(ATRP)直接检测引发剂依赖的聚合物的形成，因为最初的非荧光多环芳烃探针单体在聚合时实时显示可见的荧光。要将这种初步的概念验证应用于生物检测应用，既需要对探针荧光激活有更详细的机理了解，也需要能够直接从生物分子表面引发荧光聚合。在这里，我们表明，简单的单体氢化，独立于聚合，揭示了探针荧光，支持了共价烯醇键在荧光探针猝灭和随后的荧光激活中的关键作用。接下来，我们通过对模型蛋白质--牛血清白蛋白(BSA)的蛋白质-引发剂偶联物的表面偶联和表征，展示了生物正交的、蛋白质引发的荧光ATRP。来自引发剂修饰的蛋白质的荧光ATRP允许实时可视化聚合物的形成，而来自未修饰的BSA对照的背景荧光可以忽略不计。我们进一步探讨了这种荧光ATRP分析的生物正交性，通过评估在复杂的生物环境中聚合物的形成，添加了胎儿牛血清。综上所述，我们展示了水基荧光ATRP作为一种稳健的、生物正交的方法通过实时荧光激活进行生物分子引发聚合的潜力。
Fluorogenic atom transfer radical polymerization (ATRP) directly detects initiator-dependent polymer formation, as initially non-fluorescent polycyclic aromatic probe monomers reveal visible fluorescence upon polymerization in real time. Advancement of this initial proof-of-concept toward biodetection applications requires both a more detailed mechanistic understanding of probe fluorescence activation, and the ability to initiate fluorogenic polymerization directly from a biomolecule surface. Here, we show that simple monomer hydrogenation, independent of polymerization, reveals probe fluorescence, supporting the critical role of covalent enone attachment in fluorogenic probe quenching and subsequent fluorescence activation. We next demonstrate bioorthogonal, protein-initiated fluorogenic ATRP by the surface conjugation and characterization of protein–initiator conjugates of a model protein, bovine serum albumin (BSA). Fluorogenic ATRP from initiator-modified protein allows for real-time visualization of polymer formation with negligible background fluorescence from unmodified BSA controls. We further probe the bioorthogonality of this fluorogenic ATRP assay by assessing polymer formation in a complex biological environment, spiked with fetal bovine serum. Taken together, we demonstrate the potential of aqueous fluorogenic ATRP as a robust, bioorthogonal method for biomolecular-initiated polymerization by real-time fluorescence activation.