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Abstract

Immunology and Infection

高产细胞外囊泡制剂的纯化远离病毒

Published: September 12th, 2019

DOI:

10.3791/59876

1Laboratory of Molecular Virology, School of Systems Biology, George Mason University, 2American Type Culture Collection (ATCC), 3ATCC Cell Systems, 4Ceres Nanosciences, Inc, 5Department of Immunology and Nano-medicine, Herbert Wertheim College of Medicine, Florida International University
* These authors contributed equally

Abstract

当今细胞外囊泡 (EV) 研究领域的主要障碍之一是在病毒感染环境中实现纯化 EV 制剂的能力。提出的方法旨在将EV与病毒分离(即HIV-1),与传统的超离心方法相比,具有更高的效率和产量。我们的协议包含三个步骤:EV沉淀、密度梯度分离和粒子捕获。下游测定(即西斑点和PCR)可以直接在颗粒捕获后运行。此方法比其他隔离方法(即超离心)更有利,因为它允许使用最小起始体积。此外,它比需要多个超离心步骤的替代 EV 隔离方法更便于用户使用。然而,由于很难从粒子中洗去完整的EV,因此所呈现的方法在功能EV测定的范围上受到限制。此外,这种方法是针对严格的基于研究的设定,在商业上不可行。

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