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

doi:10.1021/acs.analchem.2c02378

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

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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 language	English (US)
Pages (from-to)	11723-11727
Number of pages	5
Journal	Analytical Chemistry
Volume	94
Issue number	34
DOIs	https://doi.org/10.1021/acs.analchem.2c02378
State	Published - Aug 30 2022

ASJC Scopus subject areas

Analytical Chemistry

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Kostelic, M. M., Ryan, J. P., Brown, L. S., Jackson, T. W., Hsieh, C. C., Zak, C. K., Sanders, H. M., Liu, Y., Chen, V. S., Byrne, M., Aspinwall, C. A., Baker, E. S., & Marty, M. T. (2022). Stability and Dissociation of Adeno-Associated Viral Capsids by Variable Temperature-Charge Detection-Mass Spectrometry. Analytical Chemistry, 94(34), 11723-11727. https://doi.org/10.1021/acs.analchem.2c02378

Kostelic, MM, Ryan, JP, Brown, LS, Jackson, TW, Hsieh, CC, Zak, CK, Sanders, HM, Liu, Y, Chen, VS, Byrne, M, Aspinwall, CA, Baker, ES & Marty, MT 2022, 'Stability and Dissociation of Adeno-Associated Viral Capsids by Variable Temperature-Charge Detection-Mass Spectrometry', Analytical Chemistry, vol. 94, no. 34, pp. 11723-11727. https://doi.org/10.1021/acs.analchem.2c02378

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

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.",

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

note = "Funding Information: We thank Jacob McCabe and Dave Russell for sharing the design and providing parts for the VT-ESI source. Also, we thank Mike Senko, Kristina Srzentic, Maria Reinhardt-Szyba, Kyle Fort, and Alexander Makarov for support on the UHMR Q-Exactive HF Orbitrap. This work was funded by the National Science Foundation (CHE-1845230 to M.T.M.) and (CBET-2003297 to C.A.A) and the UA Undergraduate Research in Biology Program. J.P.R. and E.S.B. acknowledge support from NIIMBL and NIST. Publisher Copyright: {\textcopyright} 2022 American Chemical Society. All rights reserved.",

year = "2022",

month = aug,

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doi = "10.1021/acs.analchem.2c02378",

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AU - Kostelic, Marius M.

AU - Ryan, Jack P.

AU - Brown, Levi S.

AU - Jackson, Tyler W.

AU - Hsieh, Chih Chieh

AU - Zak, Ciara K.

AU - Sanders, Henry M.

AU - Liu, Yang

AU - Chen, Victor Shugui

AU - Byrne, Michael

AU - Aspinwall, Craig A.

AU - Baker, Erin S.

AU - Marty, Michael T.

N1 - Funding Information: We thank Jacob McCabe and Dave Russell for sharing the design and providing parts for the VT-ESI source. Also, we thank Mike Senko, Kristina Srzentic, Maria Reinhardt-Szyba, Kyle Fort, and Alexander Makarov for support on the UHMR Q-Exactive HF Orbitrap. This work was funded by the National Science Foundation (CHE-1845230 to M.T.M.) and (CBET-2003297 to C.A.A) and the UA Undergraduate Research in Biology Program. J.P.R. and E.S.B. acknowledge support from NIIMBL and NIST. Publisher Copyright: © 2022 American Chemical Society. All rights reserved.

PY - 2022/8/30

Y1 - 2022/8/30

N2 - 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.

AB - 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.

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

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