Nanopore Methods for Determining Capsid Viability

测定衣壳活力的纳米孔方法

• 批准号: 10693909
• 负责人: Kimberly Ritola
• 金额: $ 39.81万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 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://reporter.nih.gov/project-details/10693909
• 关键词: Nanopore; Methods; Determining; Capsid; Viability

Nanopore Methods for Determining Capsid Viability ABSTRACT AAV (adeno-associated virus) vectors are quickly becoming an important therapeutic tool for treatment of a variety of diseases, resulting in a significant increase in the number of applications to the FDA for approvals for treatment of various diseases. Precise characterization of these AAVs is extremely important for process control and determination of the effectiveness of the final product for both production efficiency and regulatory/quality reasons. We have developed nano-fluidic, synthetic pore-based sensors capable of detecting and quantifying various nanoparticles. These existing sensors can be modified for application to capsid characterization. Demonstrated applications of this technology include DNA and RNA sequencing, SARS-CoV-2 virus detection from saliva (including viral load and infectivity classification) using resistive pulse sensing (RPS), and analysis of ligase detection reaction (LDR) products utilizing Time-of-Flight (TOF) capabilities. Specifically for this project, we will use the nanopore technology to detect capsids and characterize them as either full or empty (Aim 1) and we will extend the sensor capabilities to address high-throughput strategies for inline analysis (Aim 2). Finally, we will implement the sensors into AAV production, validate their responses against established QCAs and use sensor data to optimize production processes (Aim 3). We expect these developments to result in sensor systems that will help to improve AAV production efficiencies, improve final product infectivity, and provide important quality assurance data.

测定衣壳活力的纳米孔方法