Stability and Dissociation of Adeno-Associated Viral Capsids by Variable Temperature-Charge Detection-Mass Spectrometry
Stability and Dissociation of Adeno-Associated Viral Capsids by Variable Temperature-Charge Detection-Mass Spectrometry
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DOI：
10.1021/acs.analchem.2c02378
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发表时间：
2022-08-18
影响因子：
7.4
通讯作者：
Marty, Michael T.
中科院分区：
文献类型：
--
作者：
Kostelic, Marius M.;Ryan, Jack P.;Marty, Michael T.
腺相关病毒（AAV）载体已成为基因治疗和疫苗递送系统。差示扫描荧光法或差示扫描量热法通常用于测量AAV的热稳定性，但这些全局方法无法区分同一样本中不同AAV亚群的稳定性。为了应对这一挑战，我们将电荷检测质谱 (CD-MS) 与可变温度 (VT) 电喷雾源相结合，在电喷雾之前控制溶液的温度。使用 VT-CD-MS，我们测量了空衣壳和填充衣壳的热稳定性。我们发现，充满的 AAV 在完全解离之前首先排出其货物并形成中间空衣壳。最后，我们观察到 pH 胁迫导致热稳定性大幅下降。这种新方法可以更好地表征 AAV 的热解离，从而同时测量不同亚群的稳定性和解离途径。
Adeno-associated viral (AAV) vectors have emerged as gene therapy and vaccine delivery systems. Differential scanning fluorimetry or differential scanning calorimetry is commonly used to measure the thermal stability of AAVs, but these global methods are unable to distinguish the stabilities of different AAV subpopulations in the same sample. To address this challenge, we combined charge detection-mass spectrometry (CD-MS) with a variable temperature (VT) electrospray source that controls the temperature of the solution prior to electrospray. Using VT-CD-MS, we measured the thermal stabilities of empty and filled capsids. We found that filled AAVs ejected their cargo first and formed intermediate empty capsids before completely dissociating. Finally, we observed that pH stress caused a major decrease in thermal stability. This new approach better characterizes the thermal dissociation of AAVs, providing the simultaneous measurement of the stabilities and dissociation pathways of different subpopulations.