Multiplexing AAV Serotype-Specific Neutralizing Antibodies in Preclinical Animal Models and Humans.

Area of Science:

• * Gene therapy development
• * Immunology and virology
• * Preclinical and clinical research

Background:

• * Pre-existing humoral immunity to adeno-associated virus (AAV) can impede gene therapy efficacy.
• * Natural viral infections can induce antibodies against various AAV serotypes.
• * Accurate assessment of these antibodies is crucial for patient selection and therapy optimization.

Purpose of the Study:

• * To develop and validate a rapid and accurate assay for quantifying AAV-specific neutralizing antibodies.
• * To assess pre-existing immunity across multiple AAV serotypes (AAV1-AAV10, AAVAnc80L65).
• * To enable informed clinical trial enrollment and personalized AAV serotype selection.

Main Methods:

• * Application of equivalent infection conditions for all tested AAV serotypes.
• * Validation of the assay using samples from human donors.
• * Testing in established preclinical models: dogs and non-human primates (NHPs).

Main Results:

• * Successful rapid and accurate evaluation of neutralizing antibody titers for individual subjects.
• * Demonstrated assay performance in both human and large animal models.
• * Established a method applicable to a wide range of AAV serotypes.

Conclusions:

• * The validated assay provides a critical tool for assessing AAV immunity.
• * Enables precise patient stratification for AAV-based gene therapies.
• * Facilitates the individualized selection of optimal AAV serotypes for clinical application.