Multimodal Label-Free Nanosensor for Single Virus Characterization and Content Analysis

用于单一病毒表征和内容分析的多模式无标记纳米传感器

基本信息

  • 批准号:
    10641529
  • 负责人:
  • 金额:
    $ 48.96万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2023
  • 资助国家:
    美国
  • 起止时间:
    2023-09-25 至 2027-08-31
  • 项目状态:
    未结题

项目摘要

PROJECT SUMMARY Recently virus particles have gained immense attention for their potential as transformative therapeutic carriers. For example, the FDA-approved biologic Luxturna is an adeno-associated virus (AAV) used to carry genetic materials to treat hereditary blindness. In drug delivery, quality control is very important and the percentage of empty and part-filled capsids in such a sample needs to be determined accurately to avoid possible catastrophic effects of overdosing. While there are existing techniques like ELISA and qPCR that are used for this purpose, blind studies have shown alarmingly high intra-sample standard deviations. We propose a simpler yet more effective technique to discriminate AAVs depending on their cargo content – a plasmonic nanopore sensor with automated recapture capability for electrical sensing, aligned to operate in tandem with an optical trapping system. Viruses are soft nanoparticles and the deformability of AAV capsids depends on their cargo content. We will implement our bimodal virus characterization platform through our three Specific Aims: (1a) Characterize deformation dynamics of single AAVs during translocations through solid-state nanopores; (1b) Recapture each virus after translocation and automate a recapture protocol; (2) Capture single AAVs by self-induced back-action (SIBA) actuated nanopore electrophoresis (SANE) optical trapping in tandem with electrical recapturing; (3) Achieve robust classification of AAVs based on their cargo load by applying machine learning to optical-electrical signals that depend on virus deforamability during experiments. First, we will track the voltage-induced deformability of three AAV samples of different cargo contents: filled with ssDNA (AAVssDNA), dsDNA (AAVdsDNA), and empty capsids (AAVempty) through a range of voltages. Each type of AAV is expected to induce a unique and reproducible change in relative translocation current versus applied voltage. A narrow voltage range that shows the best discrimination will be applied on different percentage mixtures of AAVssDNA+AAVempty and AAVdsDNA+AAVempty. Conditions will be optimized to get the best discrimination between cargo-filled AAVs and the empty/part-filled capsid populations. Once this step is successful, we will incorporate single/multiple recapture capability to the platform to gain statistically sound data per AAV particle. Next, a gold double nanohole structure will be fabricated on top of the nanopore to optically trap AAVs by SIBA force, which will further deform AAVs and slow down their translocation. Importantly, optical-electrical tandem sensing will allow characterization of single AAV size deformations decoupled from charge effects. For automatic classification of viruses, we will develop a probabilistic (Bayesian) machine learning model with multiple data segments from optical-electrical signals of single AAVs for very high accuracy predictions. The proposed machine leanring assisted electrical- optical sensing will detect the fraction of viral particles (e.g., the AAVdsDNA fraction) from a complex AAV mixture while using only pico- to nano-molar concentrations, in contrast to current analytical ultracenrifugation technologies that use up significant amounts of an AAV preparation in the quality assurance step.
项目总结

项目成果

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MinJun Kim其他文献

MinJun Kim的其他文献

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{{ truncateString('MinJun Kim', 18)}}的其他基金

Nanopore Force Spectroscopy and Sorting of Vesicles at Nanoscale
纳米级囊泡的纳米孔力光谱和分选
  • 批准号:
    10158532
  • 财政年份:
    2020
  • 资助金额:
    $ 48.96万
  • 项目类别:
Nanopore Force Spectroscopy and Sorting of Vesicles at Nanoscale
纳米级囊泡的纳米孔力光谱和分选
  • 批准号:
    9979218
  • 财政年份:
    2020
  • 资助金额:
    $ 48.96万
  • 项目类别:
Nanopore Force Spectroscopy and Sorting of Vesicles at Nanoscale
纳米级囊泡的纳米孔力光谱和分选
  • 批准号:
    9292313
  • 财政年份:
    2016
  • 资助金额:
    $ 48.96万
  • 项目类别:
Nanopore Force Spectroscopy and Sorting of Vesicles at Nanoscale
纳米级囊泡的纳米孔力光谱和分选
  • 批准号:
    9340833
  • 财政年份:
    2016
  • 资助金额:
    $ 48.96万
  • 项目类别:

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