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Anal Chem. 2016 Jul 5;88(13):6718-25. doi: 10.1021/acs.analchem.6b00883. Epub 2016 Jun 16.

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

Author information

1
Department of Chemistry, Indiana University , Bloomington, Indiana 47405, United States.
2
Gene Therapy Center, University of North Carolina , Chapel Hill, North Carolina 27599, United States.

Abstract

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.

PMID:
27310298
PMCID:
PMC6537880
DOI:
10.1021/acs.analchem.6b00883
[Indexed for MEDLINE]
Free PMC Article

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