Stability and Dissociation of Adeno-Associated Viral Capsids by Variable Temperature-Charge Detection-Mass Spectrometry

Marius M. Kostelic, Jack P. Ryan, Levi S. Brown, Tyler W. Jackson, Chih Chieh Hsieh, Ciara K. Zak, Henry M. Sanders, Yang Liu, Victor Shugui Chen, Michael Byrne, Craig A. Aspinwall, Erin S. Baker, Michael T. Marty

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

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.

Original languageEnglish (US)
JournalAnalytical Chemistry
DOIs
StatePublished - Aug 30 2022

ASJC Scopus subject areas

  • Analytical Chemistry

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