Rapid At-Line AAVX Affinity HPLC: Enabling Process Analytical Technology for Bioprocess Development of...

Area of Science:

• Biotechnology and Gene Therapy
• Process Analytical Technology (PAT)
• Analytical Chemistry

Background:

• Recombinant adeno-associated virus (rAAV) vectors show therapeutic promise but face production challenges.
• Process Analytical Technology (PAT) can enhance rAAV production, but lacks suitable analytical tools.
• Existing methods struggle with consistent and efficient rAAV production.

Purpose of the Study:

• To evaluate a novel AAVX affinity-based high-performance liquid chromatography (HPLC) method as an at-line PAT tool.
• To determine capsid titer and percentage of filled capsids during rAAV production.
• To demonstrate the method's utility for improved process monitoring and control.

Main Methods:

• Development and application of an AAVX affinity-based HPLC method.
• At-line analysis of rAAV samples during upstream and downstream processing (DSP).
• Quantification of capsid titer and filled capsid percentages.

Main Results:

• The AAVX HPLC method provides fast and robust determination of capsid titer and filled capsid percentages.
• The method effectively analyzes challenging samples from fermentation and DSP stages.
• Demonstrated improved process monitoring and control capabilities.

Conclusions:

• The developed AAVX HPLC method serves as a valuable at-line PAT tool for rAAV production.
• The method's versatility across different rAAV serotypes and payloads supports streamlined process development.
• This contributes to high-quality, cost-efficient production of rAAV-based gene therapies.