University of Minnesota Background
Adeno-associated virus (AAV) vectors are a common platform for in vivo gene therapy applications and have been used successfully in clinical trials in patients with blood diseases and blindness. The presence of pre-existing neutralizing antibodies in about 50% of patients significantly decreased the clinical utility of AAV vectors. The prevailing assays used to screen for AAV neutralizing antibodies are not standardized, and can produce unreliable data depending on the AAV serotype.
Researchers at the University of Minnesota have developed a fast and reliable method to evaluate pre-existing AAV neutralizing antibodies for most of the commonly used AAV serotypes. The resulting assay can be used with virtually all AAV serotypes, requires small amounts of AAV and can produce reliable data in 24 hours.
The technology leverages a drug, Dorsomorphin (also known as Compound C), that drastically increases infection of HEK293 cells by AAV vectors. Using AAV vectors containing expression cassettes for luciferase, the increased infection rate provides a broad, measurable dynamic range to assess the infection rates of cells. This sensitized infection-rate assay facilitates reliable determination of the AAV-neutralization titers or various antibodies and/or serum. The research team optimized the assay and has shown it to be effective for measuring neutralization of three clinically relevant AAV serotypes in micel.
Modifiers of Adeno-Associated Virus-Mediated Gene Expression in Implication for Serotype-Universal Neutralizing Antibody Assay
Stage of Development
Technology Readiness Level (TRL): 3‑4
Assay tested with ten different clinically relevant AAV serotypes in a mouse model
Sensitive assay to assess neutralization of AAVs: A major strength of the assay is its enhanced dynamic range, facilitating sensitive detection of AAV neutralizing antibodies.
Simple assay with replicable results: The AAV neutralization antibody assay uses readily available reagents and well know techniques, relieving the need for a high level of expertise and increasing reproducibility.
Fast and economical: Assay can be completed in 24 hours without the requiring costly reagents.
Clinical trials: determine patient AAV neutralizing antibody status
Detect seroconversion in patients treated with AAV vectors
This technology is now available for: