High-Throughput Purification and Quantification of Bionanoparticles on Capillary-Channeled Fiber Stationary Phases

毛细管通道纤维固定相上生物纳米颗粒的高通量纯化和定量

• 批准号: 2107882
• 负责人: Richard Marcus
• 金额: $ 55万
• 依托单位: Clemson University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2107882&HistoricalAwards=false
• 关键词: Throughput; Purification; Quantification; Bionanoparticles; Capillary

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor R. Kenneth Marcus and his group at Clemson University are working to prepare, characterize, and implement a unique format of fiber capillary-channeled polymers (C-CP) to facilitate the isolation, purification and quantification of biological and synthetic nanoparticles. Target systems include exosomes, a class of extracellular vesicles (EVs) that are integral components in intracellular communication, and thus are involved in normal homeostatic function, but also in the the evolution of abnormal biological function; thus there are long term implications of the studies being performed herein for biomedical science. The applications under study employ a hydrophobic interaction chromatography (HIC) protocol on polyester C-CP fiber columns, a system designed to probe aspects of fundamental biochemistry. The overall goal of the studies is to produce efficient, selective, and economical separations. This work is interdisciplinary, involving collaborative efforts among research groups in chemistry, bioengineering, mathematical, and materials sciences. If successful, the project may result in new commercial ventures in the textile and biopharmaceutical industries in South Carolina. Efforts related to developing a diverse workforce include integration of research topics into relevant coursework.The vast majority of separation/purification strategies for biological and synthetic nanoparticles involve physical means such as ultracentrifugation, ultrafiltration, and size-exclusion methods. Use of C-CP fibers to affect these separations rests on the ability to differentiate nanoparticles based upon chemical functionality, whose interaction strengths are determined in part based on particle size. The C-CP fiber platform can be implemented across a number of formats including spin-down micropipette tips and microbore columns. As a result, relevant problems in fundamental biochemistry, clinical analysis, and process analytical chemistry can be addressed. In-line particle sizing via multi-angle light scattering (MALS) provides both qualitative and quantitative information about eluted nanoparticles/vesicles. Surface modifications employing immobilized ligands can target selective capture of nanoparticles, complemented by selective immunolabelling strategies for species-specific assays. Multidimensional separations offer another innovative aspect for efficient characterization of nanoparticles from complex systems. In all cases, performance metrics will be compared to those available from commercial columns and standard methods. Development of practical methods will be augmented with fundamental studies directed at understanding physico-chemical processes at the microscopic level.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.

在化学系化学测量和成像项目的支持下，R。Kenneth Marcus和他在克莱姆森大学的研究小组正在努力制备、表征和实施一种独特的纤维毛细管通道聚合物（C-CP）形式，以促进生物和合成纳米颗粒的分离、纯化和定量。目标系统包括外泌体，这是一类细胞外囊泡（EV），它们是细胞内通讯的组成部分，因此参与正常的稳态功能，也参与异常生物功能的演变;因此，本文正在进行的研究对生物医学科学具有长期影响。 正在研究的应用程序采用了疏水相互作用色谱（HIC）协议的聚酯C-CP纤维柱，一个系统，旨在探测方面的基础生物化学。研究的总体目标是产生有效的、选择性的和经济的分离。这项工作是跨学科的，涉及化学，生物工程，数学和材料科学研究小组之间的合作努力。如果成功，该项目可能会导致新的商业企业在纺织和生物制药行业在南卡罗来纳州。与发展多样化的劳动力相关的努力包括将研究课题整合到相关的课程中。生物和合成纳米颗粒的绝大多数分离/纯化策略涉及物理手段，如超离心，超滤和尺寸排阻方法。使用C-CP纤维来影响这些分离取决于基于化学官能度区分纳米颗粒的能力，其相互作用强度部分地基于粒度来确定。C-CP光纤平台可以跨多种形式实现，包括自旋下降微量移液器尖端和微孔柱。 因此，基础生物化学，临床分析和过程分析化学中的相关问题可以得到解决。通过多角度光散射（MALS）进行的在线粒度测定提供了关于洗脱的纳米颗粒/囊泡的定性和定量信息。 采用固定化配体的表面修饰可以靶向选择性捕获纳米颗粒，并辅以用于物种特异性测定的选择性免疫标记策略。多维分离为从复杂体系中有效表征纳米颗粒提供了另一个创新方面。在所有情况下，性能指标将与商业色谱柱和标准方法提供的指标进行比较。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Facile, generic capture and on-fiber differentiation of exosomes via confocal immunofluorescence microscopy using a capillary-channeled polymer fiber solid-phase extraction tip

• DOI: 10.1039/d2sd00007e
• 发表时间: 2022-05-19
• 期刊: SENSORS & DIAGNOSTICS
• 作者: Jackson, Kaylan K.;Powell, Rhonda R.;Marcus, R. Kenneth
• 通讯作者: Marcus, R. Kenneth

Determination of the Loading Capacity and Recovery of Extracellular Vesicles Derived from Human Embryonic Kidney Cells and Urine Matrices on Capillary-Channeled Polymer (C-CP) Fiber Columns

• DOI: 10.3390/separations9090251
• 发表时间: 2022-09
• 期刊: Separations
• 影响因子: 2.6
• 作者: Lacey S. Billotto;Kaylan K. Jackson;R. Marcus

A rapid capillary-channeled polymer (C-CP) fiber spin-down tip approach for the isolation of plant-derived extracellular vesicles (PDEVs) from 20 common fruit and vegetable sources

• DOI: 10.1016/j.talanta.2022.123779
• 发表时间: 2022-08-19
• 期刊: TALANTA
• 影响因子: 6.1
• 作者: Jackson, Kaylan K.;Mata, Carolina;Marcus, R. Kenneth
• 通讯作者: Marcus, R. Kenneth

Isolation, purification and initial RNA sequence analysis of seminal fluid exosomes between pregnant and non-pregnant intrauterine insemination pregnancies

妊娠与非妊娠宫内授精妊娠精液外泌体的分离、纯化和初始RNA序列分析

• DOI: 10.1016/j.rbmo.2020.08.019
• 发表时间: 2020
• 期刊: Reproductive BioMedicine Online
• 影响因子: 4
• 作者: Chosed, R.J.;Polley, E.;Pike, J.F.W.;Huang, S.;Zimmerman, S.;Bruce, T.F.;Marcus, R.K.;Roudebush, W.E.
• 通讯作者: Roudebush, W.E.

Comparison of the separation of proteins of wide‐ranging molecular weight via trilobal polypropylene capillary‐channeled polymer fiber, commercial superficiously porous, and commercial size exclusion columns

通过三叶聚丙烯毛细管通道聚合物纤维、商用表面多孔柱和商用尺寸排阻柱分离宽范围分子量蛋白质的比较

• DOI: 10.1002/jssc.202100891
• 发表时间: 2022
• 期刊: Journal of Separation Science
• 影响因子: 3.1
• 作者: Huang, Sisi;McClain, Ray T.;Marcus, Richard Kenneth
• 通讯作者: Marcus, Richard Kenneth