Stanley Feeney, Marissa Morales, Galen Arnold, Wynter Paiva, Eva Rose M. Balog and Jeffrey Mark Halpern*,
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引用次数: 0
摘要
弹性蛋白样聚合物(ELPs)已用于各种生物医学应用,包括药物输送和组织支架。elp由于其可调的低临界溶液温度和可调的结构而适用于不同的应用。然而,尽管对水溶液中的elp进行了充分的表征,但对表面上elp的表征却进行了较少的探索。例如,对表面进行ELP修改时不一致的来源尚未得到详细的探索。大型大分子的表面修饰常常存在再现性差和测量结果不一致的问题。我们开发并优化了一种用ELPs修饰金电极表面的方法,该方法利用巯基-金相互作用通过n端附近的单个半胱氨酸残基。通过电化学阻抗谱法测量了改性参数对表面可重复性电荷转移电阻的影响。最终优化的表面改性参数为0.0125 mg/mL ELP, 4°C, 3.5 mM TCEP, pH 7.4的超高纯水,无二甲亚砜或其他助表面活性剂处理,30分钟。通过硫醇还原法测定了半胱氨酸修饰金的相对量与ELP溶液浓度的关系。利用这些数据,可重复性差的来源被证实是非特异性聚合物相互作用。
Elastin-like polymers (ELPs) have been used for a variety of biomedical applications, including drug delivery and tissue scaffolding. ELPs are useful due to their adjustable lower critical solution temperature and tunable structure for different applications. However, despite ample characterization of ELPs in aqueous solutions, the characterization of ELPs on surfaces is less well explored. For example, sources of inconsistency in ELP modification to surfaces have yet to be explored in detail. Surface modifications of large macromolecules often suffer from poor reproducibility and inconsistent measurements. We developed and optimized a method for modifying a gold electrode surface with ELPs using a thiol-gold interaction through a single cysteine residue near the N-terminus. The modification parameters were tuned for reproducible charge-transfer resistance of the surface, as measured by electrochemical impedance spectroscopy. The final optimized surface modification parameters, without dimethyl sulfoxide or other cosurfactant treatment, are 0.0125 mg/mL ELP for 30 min at 4 °C in 3.5 mM TCEP in ultrahigh-purity water at pH 7.4. The relative amount of cysteine modified to gold versus ELP solution concentration was determined via thiol reduction. Using these data, the source of poor reproducibility was confirmed to be nonspecific polymer interactions.
期刊介绍:
ACS Measurement Science Au is an open access journal that publishes experimental computational or theoretical research in all areas of chemical measurement science. Short letters comprehensive articles reviews and perspectives are welcome on topics that report on any phase of analytical operations including sampling measurement and data analysis. This includes:Chemical Reactions and SelectivityChemometrics and Data ProcessingElectrochemistryElemental and Molecular CharacterizationImagingInstrumentationMass SpectrometryMicroscale and Nanoscale systemsOmics (Genomics Proteomics Metabonomics Metabolomics and Bioinformatics)Sensors and Sensing (Biosensors Chemical Sensors Gas Sensors Intracellular Sensors Single-Molecule Sensors Cell Chips Arrays Microfluidic Devices)SeparationsSpectroscopySurface analysisPapers dealing with established methods need to offer a significantly improved original application of the method.