2022年1月5日 · General discrepancies on PTM abundance levels can be explained by differences in expression systems or overall downstream processing. Nevertheless, it is important to highlight that our workflow using pepsin digestion provides 100% sequence coverage in 30 min, giving a better insight into AAV5 sequence and associated PTMs.

2024年9月12日 · Potential N-linked glycosylation is another important PTM to monitor on AAV vectors as it can affect the tropism, immunogenicity, and intracellular trafficking of the viral vector. 42 Studies have reported conflicting evidence regarding glycosylation importance in AAV2 and …

Our analysis revealed a total of 52 post‐translational modifications in AAV2‐AAVrh10 capsids, including ubiquitination (17%), glycosylation (36%), phosphorylation (21%), SUMOylation (13%) and acetylation (11%).

2024年12月15日 · Opposite charge profile shifts observed for thermally stressed AAV8 and AAV1. An optimized one-pot tryptic digestion protocol developed for AAV peptide mapping. The location of modifications largely impacts the AAV retention on AEX. A unique PTM specific to each AAV serotype is mainly responsible for the observed charge profile shift.

氨基酸序列确认或肽图（Peptide Mapping）序列覆盖度是确认AAV产品衣壳蛋白一级结构和理论设计符合的重要表征手段。 在氨基酸序列确认过程中，借助工作站氨基酸序列突变和翻译后修饰的计算能力，可以得出具参考价值的氨基酸位点突变情况和翻译后修饰（如N-端乙酰化，甲基化，磷酸化，氧化等）情况。 N-糖修饰 在AAV病毒进入、组织选择和传染性中起着重要作用。 同时，N-糖修饰也是衣壳修饰的重要部分。 N-糖糖型和糖基化位点的表征也是衣壳一级结构表征的重 …

质谱(Mass spectrometry，MS)是AAV的关键分析工具之一，可以在AAV开发过程中直接、准确地评估AAV产品的质量。 早期，人们尝试使用傅里叶变换离子回旋共振质谱来表征衣壳蛋白的糖基化，以及四极杆飞行时间质谱法(QTOF)来鉴定不同AAV的血清型。

此外，根据 Mary B. 等人对 10 种 AAV 血清型进行的系统分析，重组 AAV1 (rAAV1) 未发现任何可检测到的翻译后修饰 (PTM)。 它是第一个被批准用于基因治疗的病毒载体。

Characterization of AAV capsid proteins (VPs) and their post-translational modifications (PTMs) have become a critical attribute monitored to evaluate product quality. Liquid chromatography–mass spectrometry (LC–MS) analysis of intact AAV VPs provides both quick and reliable serotype identification as well as proteoform information on each VP.

Ideally, AAV production should be with full control of AAV structure and genetic payload. Therefore, robust, efficient, and low material consuming methods are essential to characterize AAVs. Here, we use two emerging single-molecule techniques, mass photometry and Orbitrap-based charge-detection mass spectrometry, and show how they may ...

2021年7月22日 · The developed application of pepsin provided 100% sequence coverage of AAV2 capsid proteins and demonstrated the potential for PTM analysis and more in-depth structural and quantitative characterization of AAVs than the conventional trypsin-based approaches showed.