衣壳
腺相关病毒
原子力显微镜
病毒学
病毒
显微镜
遗传增强
生物物理学
化学
基因传递
纳米技术
材料科学
生物
基因
物理
光学
重组DNA
载体(分子生物学)
遗传学
作者
Yu Ri Nam,Helen H. Ju,Jeehee Lee,Daiheon Lee,Yoojin Kim,Sung Jin Lee,Hong Kee Kim,Jae‐Hyung Jang,Haeshin Lee
出处
期刊:Langmuir
[American Chemical Society]
日期:2023-05-02
卷期号:39 (19): 6740-6747
被引量:2
标识
DOI:10.1021/acs.langmuir.3c00241
摘要
Recently, miraculous therapy approaches involving adeno-associated virus (AAV) for incurable diseases such as spinal muscular atrophy and inherited retinal dysfunction have been introduced. Nonreplicative, nonpathogenic, low rates of chromosome insertional properties and the existence of neutralizing antibodies are main safety reasons why the FDA approved its use in gene delivery. To date, AAV production always results in a mixture of nontherapeutic (empty) and therapeutic (DNA-loaded) full capsids (10–98%). Such existence of empty viral particles inevitably increases viral doses to human. Thus, the rapid monitoring of empty capsids and reducing the empty-to-full ratio are critical in AAV science. However, transmission electron microscopy (TEM) is the primary tool for distinguishing between empty and full capsids, which creates a research bottleneck because of instrument accessibility and technical difficulty. Herein, we demonstrate that atomic force microscopy (AFM) can be an alternative tool to TEM. The simple, noncontact-mode imaging of AAV particles allows the distinct height difference between full capsids (∼22 nm) and empty capsids (∼16 nm). The sphere-to-ellipsoidal morphological distortion observed for empty AAV particles clearly distinguishes them from full AAV particles. Our study indicates that AFM imaging can be an extremely useful, quality-control tool in AAV particle monitoring, which is beneficial for the future development of AAV-based gene therapy.
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