RAPID: Determination of the cross-over frequency of SARS-CoV-2 for rapid identification concentration

RAPID：测定 SARS-CoV-2 的交叉频率以快速识别浓度

• 批准号: 2031741
• 负责人: Woo Jin Chang
• 金额: $ 19.32万
• 依托单位: University of Wisconsin-Milwaukee
• 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: 2020-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2031741&HistoricalAwards=false
• 关键词: RAPID; Determination; cross; over; frequency

The University of Wisconsin-Milwaukee received an award to investigate an electrical trapping technique to rapidly concentrate SARS-CoV-2 virus to produce highly pure samples for the development of vaccines and therapeutics. Currently, purification through multiple in-series applications of filtration and centrifugation require up to two days to prepare virus samples. Unfortunately, the purity and viability of the separated viruses are still low after being collected by conventional methods. In this research, a correlation between the virus, nanoparticle size, and electrical dielectrophoretic trapping conditions is investigated to determine the best operating conditions to concentrate 80 – 160 nm size SARS-CoV-2. The developed method will enable the rapid concentration of pure viruses from cell culture and patient samples. The concentrated samples will enhance the development of vaccines, cures, and rapid diagnostic methods for infectious diseases by rapidly supplying highly concentrated pure virus samples. Thus, the research will contribute to improve the public safety related to infectious diseases and economy in the diagnostics and therapeutics area. The developed method is also applicable to the concentration of biomolecules, including DNA, protein, cell lysed fragments and particles, in addition to viruses. Thus, this research will give a substantial improvement on the rapid characterization and applications of molecules in various disciplines, such as virology, genomics, proteomics, diagnosis, forensic science, in addition to vaccine and therapeutics development. The novel dielectrophoretic nanoparticle manipulating method developed in this research can rapidly concentrate viruses into highly pure samples from virus-infected cell media and samples from patients. A charge polarization happens to a dielectric particle when the particle is exposed to an unevenly distributed electric field. The particle will move to either a dense or sparse electric field area depending on the given condition, such as electrical potential, frequency, and permittivities of the particle and medium. The direction of particle movement is inverted depending on the frequency; this is called cross-over frequency. The cross-over frequency varies depending on the characteristics of the particles, such as components, structure, charge, etc. In this research, the cross-over frequency of different bio-nano-particles, ranged from 60 to 500 nm size, will be investigated using heat-treated SARS-CoV-2 and nanoparticles. Initially, the cross-over frequency of different size nano-particles will be determined using dielectrophoretic traps, by modifying the applied potential, frequency, and particle size. Then, the same operating condition screening strategy will be applied to heat-treated SARS-CoV-2. Purity and efficiency of the isolated nano-particles and SARS-CoV-2 will be determined using the developed conditions. The developed method will circumvent the drawbacks that exist with the current serial treatment method used to separate virus, which are: time-intensive, laborious, complicated, and less effective. Consequently, this research will significantly enhance the understandings of the structures and characteristics of bio-nano-particles, such as live viruses, by supplying higher purity samples rapidly. Obtained results will be rapidly applied to other viruses and particles for further characterization and detection, as well as development of applications. This RAPID award to the University of Wisconsin-Milwaukee is made by the Division of Biological Infrastructure using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.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.

威斯康星大学密尔沃基分校因研究一种电子捕获技术而获奖，该技术可快速浓缩SARS-CoV-2病毒，以生产用于疫苗和疗法开发的高纯度样本。目前，通过多个串联过滤和离心法进行纯化需要长达两天的时间来准备病毒样本。遗憾的是，用常规方法收集病毒后，分离病毒的纯度和活力仍然较低。在这项研究中，研究了病毒、纳米颗粒大小和电介质电捕获条件之间的相关性，以确定浓缩80-160 nm大小的SARS-CoV-2的最佳操作条件。开发的方法将使从细胞培养和患者样本中快速浓缩纯病毒成为可能。浓缩的样本将通过快速提供高度浓缩的纯病毒样本，促进传染病疫苗、治疗和快速诊断方法的发展。因此，该研究将有助于提高与传染病相关的公共安全和诊疗领域的经济性。除病毒外，该方法还适用于生物分子的浓缩，包括DNA、蛋白质、细胞裂解片段和颗粒。因此，这项研究将大大提高分子的快速表征和在不同学科中的应用，如病毒学、基因组学、蛋白质组学、诊断、法医学，以及疫苗和治疗学的开发。本研究开发的新型介电泳纳米粒操作方法可以快速将病毒从感染病毒的细胞培养液和患者样本中浓缩成高纯度的样本。当介电粒子暴露在不均匀分布的电场中时，该粒子会发生电荷极化。粒子将根据给定的条件，如粒子和介质的电势、频率和介电常数，移动到密集或稀疏的电场区域。粒子运动的方向取决于频率的反转；这称为交叉频率。交叉频率取决于颗粒的组成、结构、电荷等特性。在本研究中，将使用热处理的SARS-CoV-2和纳米颗粒来研究尺寸从60到500 nm的不同生物纳米颗粒的交叉频率。最初，不同尺寸的纳米颗粒的交叉频率将通过改变施加的电位、频率和颗粒大小，使用介电泳捕获器来确定。然后，将相同的运行条件筛选策略应用于热处理的SARS-CoV-2。分离的纳米颗粒和SARS-CoV-2的纯度和效率将使用开发的条件进行测定。所开发的方法将绕过目前用于分离病毒的连续处理方法存在的缺点：耗时、费力、复杂、效率较低。因此，这项研究将通过快速提供更高纯度的样品来显著提高对生物纳米粒子(如活病毒)的结构和特性的了解。所获得的结果将迅速应用于其他病毒和颗粒的进一步表征和检测，以及应用的开发。这项对威斯康星大学密尔沃基分校的快速奖励是由生物基础设施部利用冠状病毒援助、救济和经济安全(CARE)法案的资金做出的。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。