Insights into Adeno-Associated Virus Capsid Charge Heterogeneity.

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Analytical Chemistry Pub Date : 2025-07-17 DOI:10.1021/acs.analchem.5c03104

Jiaqi Wu,Benjamin E Draper,Martin F Jarrold,Christopher D Heger

引用次数: 0

Abstract

A comprehensive workflow is described to examine three contributing factors to the charge heterogeneity of Adeno-associated viruses (AAVs) from a single sample. Intact AAV9 capsids were fractionated using imaged capillary isoelectric focusing (icIEF)-based fractionation, allowing for collection of capsids with different isoelectric points (pIs). Capsid integrity of the fractions was confirmed with analytical icIEF and charge detection mass spectrometry (CD-MS). Using capillary electrophoresis (CE) immunoassays, the capsid protein ratios and capsid protein deamidation were characterized. Additionally, to analyze ssDNA content packaged in each fraction, CE-immunoassay and high-resolution CD-MS were used. This study enhances our understanding of AAVs, by examining the contributions of its attributes to capsid charge heterogeneity.

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腺相关病毒衣壳电荷异质性研究

本文描述了一个全面的工作流程，以检查来自单个样本的腺相关病毒（aav）电荷异质性的三个影响因素。完整的AAV9衣壳采用基于成像毛细管等电聚焦（icIEF）的分离，允许收集具有不同等电点（pi）的衣壳。用icIEF和电荷检测质谱（CD-MS）证实了这些组分的衣壳完整性。采用毛细管电泳（CE）免疫分析法对衣壳蛋白比例和衣壳蛋白脱酰胺进行了表征。此外，为了分析每个部分中包装的ssDNA含量，使用ce免疫分析法和高分辨率CD-MS。本研究通过研究aav属性对衣壳电荷异质性的贡献，增强了我们对aav的理解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.