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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Pierson, EE; Keifer, DZ; Asokan, A; Jarrold, MF

Published Date

  • July 5, 2016

Published In

Volume / Issue

  • 88 / 13

Start / End Page

  • 6718 - 6725

PubMed ID

  • 27310298

Pubmed Central ID

  • 27310298

Electronic International Standard Serial Number (EISSN)

  • 1520-6882

Digital Object Identifier (DOI)

  • 10.1021/acs.analchem.6b00883

Language

  • eng

Conference Location

  • United States