Size Exclusion Chromatography-Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization.

Area of Science:

• Biotechnology
• Molecular Biology
• Virology

Background:

• Adeno-associated viruses (AAVs) are crucial for gene therapy and genome editing.
• Current AAV characterization methods struggle with impure or heterogeneous samples.
• A need exists for rapid, low-sample-consumption AAV analysis techniques.

Purpose of the Study:

• To develop a novel method for characterizing adeno-associated virus (AAV) products with improved accuracy and efficiency.
• To address the limitations of existing techniques in analyzing lower purity or heterogeneous AAV samples.
• To provide a robust quality control measure for AAV-based therapeutics.

Main Methods:

• Combined size exclusion chromatography (SEC) with mass photometry (MP) for AAV analysis.
• SEC separates monomeric AAV particles from impurities, with UV detection for virus particle concentration.
• MP quantifies the fraction of fully packaged AAVs within the total particle population.

Main Results:

• The SEC-MP method accurately determines the titer of effective, fully packaged AAVs.
• Successfully characterized AAV samples containing aggregates, incomplete genomes, and other impurities.
• Demonstrated the utility of SEC-MP for quality control of AAV products.

Conclusions:

• The SEC-MP method offers a precise and efficient approach for AAV product characterization.
• This technique is valuable for assessing the quality and purity of AAVs used in gene therapy.
• SEC-MP provides essential guidance for AAV research and downstream applications.