Enhancing sanitization for AVB Sepharose resin in AAV vector purification

Area of Science:

• Biotechnology
• Process Chemistry
• Gene Therapy Manufacturing

Background:

• Affinity chromatography is essential for capturing adeno-associated virus (AAV) vectors in gene therapy.
• High costs of affinity resins necessitate efficient cleaning and sanitization for multi-cycle use.
• Microbial contamination and product carryover are significant challenges in AAV purification.

Purpose of the Study:

• To develop and validate enhanced cleaning and sanitization protocols for affinity chromatography resins used in AAV purification.
• To identify effective reagent combinations against diverse microbial strains, including resistant species.
• To ensure multi-cycle resin usability without compromising process yield or product quality.

Main Methods:

• Evaluation of various cleaning reagent combinations, including alcohols, sodium hydroxide (NaOH), and acids.
• Testing efficacy against a broad spectrum of microbial strains, focusing on acid- and alkaline-resistant species.
• Implementation and validation of optimized pre-use and post-use cleaning strategies.

Main Results:

• A cleaning solution of 100 mM acetic acid with 2% benzyl alcohol and 10 mM NaOH with 2% benzyl alcohol proved effective.
• The developed strategy allowed for up to six column cycles with no product carryover.
• No compromise in process yield or product quality was observed with the new protocols.

Conclusions:

• The new cleaning and sanitization strategy provides effective microbial control for AAV purification using AVB Sepharose resin.
• This approach preserves resin integrity, reduces contamination risks, and lowers manufacturing costs.
• The validated protocols enhance the quality and reliability of gene therapy product manufacturing.