Do Your AAVs Contain DNA? Rapid and Sensitive Empty/Full Capsid Quantification with Simple Western

In this Application Note, we developed a novel method for quantifying the DNA content of AAV particles with Simple Western™, a next-generation biomolecular analytical tool that seamlessly combines capillary electrophoresis and immunodetection with conventional Western blot antibodies. Here, the Simple Western method automatically separates AAV samples by Size or Charge followed by specific and sensitive detection using anti-DNA and anti-VP1/2/3 antibodies directly in the capillary for quantitative, reproducible measurements of these central AAV components. For each antibody, we identify the range in which the signal intensity is linearly related to the amount of sample loaded, resulting in a rapid and sensitive assay for accurately quantifying the ratio of % full AAVs to total AAVs in a sample (also known as the Content Ratio).