Collaborative Research: A Stacked Plasmonic Nanopore for Tether-Free Stretching and Label-Free Sensing of hSTf Dynamics and Complex Formation at Ultra-Low Concentrations

合作研究：堆叠式等离子体纳米孔，用于超低浓度下 hSTf 动力学和复杂形成的无绳拉伸和无标记传感

• 批准号: 2022374
• 负责人: MinJun Kim
• 金额: $ 28.73万
• 依托单位: Southern Methodist University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2022374&HistoricalAwards=false
• 关键词: Collaborative; Research; Stacked; Plasmonic; Nanopore

Fundamental knowledge of protein structures and their dynamic responses to stimuli or other molecules is important for many applications, including medical diagnosis and therapy. This research aims to develop a highly sensitive approach for studying the human serum transferrin protein (hSTf), which is a vital iron carrier in blood and of clinical importance. The sensing technique would allow differentiation of the free hSTf protein from the iron-bound protein and evaluation of iron deficiency or iron overload from very small blood samples. Successful development of this sensor would also enable profiling of a wide range of other proteins and biological molecules, e.g., DNA. This project offers excellent opportunities for interdisciplinary research training as it combines biochemistry, nanoengineering, photonics, and electrical engineering. The outreach efforts to K-12 schools through various programs at the Southern Methodist University and the University of Texas at Arlington help to inspire more students to pursue science, technology, engineering and mathematics (STEM) degrees.The stacked plasmonic nanosensor is based on the self-induced back-action (SIBA) actuated nanopore electrophoresis (SANE) sensing concept. The stacked nanopores are uniquely designed to enable 1) controlled trapping, releasing, and recapturing of proteins or the substrate-bound protein complexes, 2) transient deformation of the biological molecules, which can be induced by thermal effect or a combination of optical and electrical techniques, and 3) study of their deformation dynamics. The SANE concept implemented in the stacked nanopore sensor allows investigation of protein interactions at concentrations 1000-fold below the equilibrium dissociation constant in bulk solution, making this technique ultra-sensitive. An important aim of this research is the study of the properties of free-hSTf protein and the iron-bound protein complex using the SANE sensor. Optical signature profiles are established for each of the species to enable selective admission of bound complexes over unbound proteins in a mixed solution to the underlying pore. It uses symmetric (VCapture = VRecapture), followed by asymmetric (VCapture ≠ VRecapture) voltage conditions to facilitate the investigation of the strength and kinetic parameters associated with protein-substrate binding, protein relaxation times, and whether voltage-induced protein unfolding is reversible or not.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

蛋白质结构的基本知识及其对刺激或其他分子的动态反应对于许多应用（包括医学诊断和治疗）很重要。这项研究旨在开发一种高度敏感的方法来研究人血清转铁蛋白蛋白（HSTF），这是血液和临床重要性的重要铁载体。感应技术将允许将游离HSTF蛋白与铁结合的蛋白质区分开，并评估来自非常小的血液样本的铁缺乏症或铁超载。该传感器的成功开发还将能够分析多种其他蛋白质和生物分子，例如DNA。该项目为跨学科研究培训提供了绝佳的机会，因为它结合了生物化学，纳米工程，光子学和电气工程。通过南方卫理公会大学和德克萨斯大学阿灵顿分校为K-12学校提供的推广工作有助于激发更多的学生追求科学，技术，工程和数学（STEM）学位。堆叠的等离激元纳米体基于自我诱导的背光（SIBA）激活的纳米型纳米型电动电动机（SANE）的敏感性。堆叠的纳米孔的设计独特以启用1）受控捕获，释放和重新接收蛋白质或底物结合的蛋白质复合物，2）生物分子的短暂变形，可以通过热和电气技术的热效果或研究其变形动力学来诱导，或者可以通过热效果或对光学和电气技术的组合诱导。在堆叠的纳米孔传感器中实现的理智概念允许研究蛋白质相互作用的浓度低于大量溶液中等效解离常数的浓度，从而使该技术超敏感。这项研究的一个重要目的是研究使用SANE传感器对自由HSTF蛋白和铁结合蛋白质络合物的特性的研究。为每个物种建立了光学特征曲线，以使在与基础孔混合溶液中的未结合蛋白上选择性接收性复合物。它使用对称（vcapture = vRecapture），然后使用不对称（VCAPTURE≠vRecapture）电压条件，以促进强度和动力学参数的投资，与蛋白质 - 基层结合，蛋白质松弛时间相关，蛋白质松弛时间以及电压诱导的蛋白质的启用以及均具有依赖的元素。使用基金会的智力优点和更广泛的影响评估标准进行评估。

项目成果

Investigating protein translocation in the presence of an electrolyte concentration gradient across a solid‐state nanopore

研究固态纳米孔中存在电解质浓度梯度时的蛋白质易位

• DOI: 10.1002/elps.202100346
• 发表时间: 2022
• 期刊: ELECTROPHORESIS
• 影响因子: 2.9
• 作者: Saharia, Jugal;Bandara, Y. M. Nuwan D. Y.;Kim, Min Jun
• 通讯作者: Kim, Min Jun

Nanopore Data Analysis: Baseline Construction and Abrupt Change-Based Multilevel Fitting

• DOI: 10.1021/acs.analchem.1c01646
• 发表时间: 2021-08-17
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Bandara, Y. M. Nuwan D. Y.;Saharia, Jugal;Kim, Min Jun
• 通讯作者: Kim, Min Jun

Assessment of 1/f noise associated with nanopores fabricated through chemically tuned controlled dielectric breakdown.

• DOI: 10.1002/elps.202000285
• 发表时间: 2021-04
• 期刊: Electrophoresis
• 影响因子: 2.9
• 作者: Saharia J;Bandara YMNDY;Karawdeniya BI;Alexandrakis G;Kim MJ
• 通讯作者: Kim MJ

Significant reduction in the duration of transient voltage responses of a plasmonic nanopore sensor by use of a Chebyshev filter

使用切比雪夫滤波器显着减少等离子体纳米孔传感器的瞬态电压响应持续时间

• DOI: 10.1117/12.2650798
• 发表时间: 2023
• 期刊: Significant reduction in the duration of transient voltage responses of a plasmonic nanopore sensor by use of a Chebyshev filter
• 作者: Asadzadeh, Homayoun;Turpin, Scott;Renkes, Scott;Kim, Min Jun;Alexandrakis, George
• 通讯作者: Alexandrakis, George

Use of a solid‐state nanopore for profiling the transferrin receptor protein and distinguishing between transferrin receptor and its ligand protein

使用固态纳米孔分析转铁蛋白受体蛋白并区分转铁蛋白受体及其配体蛋白

• DOI: 10.1002/elps.202200147
• 发表时间: 2022
• 期刊: ELECTROPHORESIS
• 影响因子: 2.9
• 作者: O'Donohue, Matthew;Saharia, Jugal;Bandara, Nuwan;Alexandrakis, Georgios;Kim, Min Jun
• 通讯作者: Kim, Min Jun