Resolving Adeno-Associated Viral Particle Diversity With Charge Detection Mass Spectrometry

Recombinant adeno-associated viruses (AAVs) are promising vectors for human gene therapy. However, current methods for evaluating AAV particle populations and vector purity are inefficient and low resolution. Here, we show that charge detection mass spectrometry (CDMS) can resolve capsids that conta...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2016-07, Vol.88 (13), p.6718-6725
Main Authors: Pierson, Elizabeth E, Keifer, David. Z, Asokan, Aravind, Jarrold, Martin F
Format: Article
Language:English
Subjects:
Adeno-associated virus
Analytical chemistry
Capsid - chemistry
Capsid - metabolism
Capsid Proteins - chemistry
Capsid Proteins - genetics
Capsid Proteins - metabolism
Charge
Comparative analysis
Dependovirus - chemistry
Dependovirus - genetics
Dependovirus - metabolism
DNA, Viral - chemistry
DNA, Viral - metabolism
Genetic Vectors - genetics
Genetic Vectors - metabolism
Genomes
Ion velocity
Manuals
Mass Spectrometry
Mathematical analysis
Microscopy, Electron
Recombinant
Vectors (mathematics)
Viruses
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Summary:Recombinant adeno-associated viruses (AAVs) are promising vectors for human gene therapy. However, current methods for evaluating AAV particle populations and vector purity are inefficient and low resolution. Here, we show that charge detection mass spectrometry (CDMS) can resolve capsids that contain the entire vector genome from those that contain partial genomes and from empty capsids. Measurements were performed for both single-stranded and self-complementary genomes. The self-complementary AAV vector preparation appears to contain particles with partially truncated genomes averaging at half the genome length. Comparison to results from electron microscopy with manual particle counting shows that CDMS has no significant mass discrimination in the relevant mass range (after a correction for the ion velocity is taken into account). Empty AAV capsids are intrinsically heterogeneous, and capsids from different sources have slightly different masses. However, the average masses of both the empty and full capsids are in close agreement with expected values. Mass differences between the empty and full capsids for both single-stranded and self-complementary AAV vectors indicate that the genomes are largely packaged without counterions.
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/acs.analchem.6b00883