Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan

Amy Henrickson; Xiaozhe Ding; Austin G. Seal; Zhe Qu; Lauren Tomlinson; John Forsey; Viviana Gradinaru; Kazuhiro Oka; Borries Demeler

doi:10.2217/nnm-2023-0156

Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan

Amy Henrickson
, Xiaozhe Ding
, Austin G. Seal
, Zhe Qu
, Lauren Tomlinson
, John Forsey
, Viviana Gradinaru
, Kazuhiro Oka
, Borries Demeler

Chemistry and Biochemistry

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.

Original language	English
Pages (from-to)	1519-1534
Number of pages	16
Journal	Nanomedicine (London, England)
Volume	18
Issue number	22
DOIs	https://doi.org/10.2217/nnm-2023-0156
State	Published - Sep 1 2023

Funding

This work received funding from the Beckman Institute CLOVER Center at Caltech and the NIH BRAIN Initiative Armamentarium (UF1MH128336), Canada 150 Research Chairs Program (C150-2017-00015), Canada Foundation of Innovation (CFI-37589), Canadian Natural Science and Engineering Research Council (DG-RGPIN-2019-05637), Cancer Prevention and Research Institute of Texas (RP190602), NIH-NIGMS (1R01GM120600), NSF (TG-MCB070039N) and the University of Texas (TG457201). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties. Competing interests disclosure

Funders	Funder number
Canada Foundation for Innovation	CFI-37589
TG-MCB070039N
C150-2017-00015, UF1MH128336
1R01GM120600
RP190602
University of Texas at Austin	TG457201
DG-RGPIN-2019-05637

Keywords

UltraScan
adeno-associated virus
analytical buoyant density gradient equilibrium
gene therapy
multi-wavelength analytical ultracentrifugation
sedimentation velocity
viral capsid quantification
viral vector

Access to Document

10.2217/nnm-2023-0156

Cite this

@article{cd9c3f4a4ab145c38ef23b86ed537f55,

title = "Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan",

abstract = "Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.",

keywords = "UltraScan, adeno-associated virus, analytical buoyant density gradient equilibrium, gene therapy, multi-wavelength analytical ultracentrifugation, sedimentation velocity, viral capsid quantification, viral vector",

author = "Amy Henrickson and Xiaozhe Ding and Seal, \{Austin G.\} and Zhe Qu and Lauren Tomlinson and John Forsey and Viviana Gradinaru and Kazuhiro Oka and Borries Demeler",

year = "2023",

month = sep,

day = "1",

doi = "10.2217/nnm-2023-0156",

language = "English",

volume = "18",

pages = "1519--1534",

number = "22",

}

TY - JOUR

T1 - Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan

AU - Henrickson, Amy

AU - Ding, Xiaozhe

AU - Seal, Austin G.

AU - Qu, Zhe

AU - Tomlinson, Lauren

AU - Forsey, John

AU - Gradinaru, Viviana

AU - Oka, Kazuhiro

AU - Demeler, Borries

PY - 2023/9/1

Y1 - 2023/9/1

N2 - Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.

AB - Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.

KW - UltraScan

KW - adeno-associated virus

KW - analytical buoyant density gradient equilibrium

KW - gene therapy

KW - multi-wavelength analytical ultracentrifugation

KW - sedimentation velocity

KW - viral capsid quantification

KW - viral vector

UR - https://www.scopus.com/pages/publications/85176975137

U2 - 10.2217/nnm-2023-0156

DO - 10.2217/nnm-2023-0156

M3 - Article

C2 - 37877696

AN - SCOPUS:85176975137

VL - 18

SP - 1519

EP - 1534

JO - Nanomedicine (London, England)

JF - Nanomedicine (London, England)

IS - 22

ER -

Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan

Abstract

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Cite this