Aldehyde-Mediated Protein-to-Surface Tethering via Controlled Diazonium Electrode Functionalization Using Protected Hydroxylamines.
Aldehyde-Mediated Protein-to-Surface Tethering via Controlled Diazonium Electrode Functionalization Using Protected Hydroxylamines.
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使用受保护的羟胺通过受控重氮电极功能化实现醛介导的蛋白质与表面束缚。
DOI：
10.1021/acs.langmuir.9b01254
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Yates ND
中科院分区：
文献类型：
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作者：
Yates ND
我们报告了一种重氮电接枝方法，用于用醛反应性羟胺官能团对导电表面进行共价修饰，从而促进氧化还原活性（生物）分子与电极表面的连接。羟胺近单层的形成是通过邻苯二甲酰亚胺保护和肼脱保护策略实现的，该策略克服了通常使重氮表面改性复杂化的多层形成。这种表面改性策略采用电化学（电化学阻抗谱和循环伏安法）、X 射线光电子能谱和具有耗散监测的石英晶体微天平进行表征。经过如此修饰的玻璃碳、掺硼金刚石和金表面均显示可与小分子醛连接，分别产生 150-170、40 和 100 pmol cm-2 的表面覆盖率。通过在生物相容性条件（H2O 溶剂，pH 4.5，25 °C）下将高碘酸氧化辣根过氧化物酶的稀释（50 μM）溶液与功能化金表面偶联来证明生物偶联。
We report a diazonium electro-grafting method for the covalent modification of conducting surfaces with aldehyde-reactive hydroxylamine functionalities that facilitate the wiring of redox-active (bio)molecules to electrode surfaces. Hydroxylamine near-monolayer formation is achieved via a phthalimide-protection and hydrazine-deprotection strategy that overcomes the multilayer formation that typically complicates diazonium surface modification. This surface modification strategy is characterized using electrochemistry (electrochemical impedance spectroscopy and cyclic voltammetry), X-ray photoelectron spectroscopy, and quartz crystal microbalance with dissipation monitoring. Thus-modified glassy carbon, boron-doped diamond, and gold surfaces are all shown to ligate to small molecule aldehydes, yielding surface coverages of 150–170, 40, and 100 pmol cm–2, respectively. Bioconjugation is demonstrated via the coupling of a dilute (50 μM) solution of periodate-oxidized horseradish peroxidase enzyme to a functionalized gold surface under biocompatible conditions (H2O solvent, pH 4.5, 25 °C).