In the rapidly evolving field of adeno-associated virus (AAV) manufacturing, the focus is on creating well-defined, robust processing steps backed by fast, reliable, and high-throughput analytical methods to ensure consistent product quality. One of the key challenges in AAV production is the characterization of different AAV serotypes and optimizing the ratio of empty to full capsids. Sartorius BIA Separations experts provide comprehensive analytical support throughout the entire manufacturing process.
Common Challenges in AAV Analytics
- Low Resolution: Conventional chromatographic methods only observe empty and full capsids with insufficient resolution between peaks.
- Low Purity: Conventional methods for separation, such as chromatography resins, achieve only 70% purity of full capsids.
- Incomplete Separation: Earlier conventional approaches only partially separated empty and full capsids.
- Lack of Serotype Selectivity: Only a few conventional methods offer serotype selectivity when separating AAVs.
- Complex Sample Analysis: Full percentage estimation in the upstream complex samples requires a combination of analytical methods, usually PCR and ELISA.
With Cornerstone services, these common challenges in AAV manufacturing are addressed while providing a solution tailored to specific needs.
Overcoming AAV Analytical Challenges
The enhanced anion exchange (AEX) method in combination with PATfix AAV Switcher delivers a significant improvement over conventional approaches. It allows for better separation between empty and full capsids while also identifying additional subpopulations and impurities.
- Higher Resolution Between Peaks
The enhanced AEX method facilitates the detection of various AAV subpopulations, such as empty, full, and partially full capsids. Moreover, it identifies other AAV-related impurities, including aggregates and heavy or overfilled capsids.
- Improved Purity
The improved AEX method using monolithic chromatography delivers significantly purer full AAV capsids (90%-95% pure full capsids) compared to the conventional methods.
- Better Separation of Empty and Full Capsids
The new method achieves baseline separation between empty and full capsids, as well as some additional capsids, indicating the heterogeneity of AAV samples.
- Serotype Applicability
The improved AEX method applies to different AAV serotypes, such as AAV5, AAV8 and AAV9.
- Complex Sample Analysis Simplified
PATfix AAV Switcher allows the determination of the empty and full ratio in harvest samples or spent media, based on MALS and FLD signals. FLD works in combination with MALS as it has a higher sensitivity while MALS proves to be more reliable in cases of background noise in the FLD profile.
AAV Capsids Separation is Critical for Advancing Gene Therapies
Why Choose Cornerstone AAV Process Analytics Services?
By partnering with Cornerstone AAV Process Analytics Services, the following benefits are offered:
- Higher resolution between peaks for more accurate subpopulation detection.
- Significantly improved purity of full AAV capsids (up to 95%).
- Comprehensive capsid separation with insights into additional capsid types.
- Serotype versatility across AAV5, AAV8, AAV9, and more.
- Cutting-edge analytical tools for handling complex AAV samples.
Additional Resources
Read the article AAV Capsids Separation is Critical for Advancing Gene Therapies for more detailed insights
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FAQ About Services for AAV Process Analytics
The facilities are located in Ajdovščina, Slovenia. Support for process development is available in modern, fully equipped BSL 1 and 2 facilities. A virtual tour of the laboratories is available.
Analytical testing for impurity profiling and detection, AAV recovery assessments, and different AAV capsid separation in AAV DSP samples is offered. Method development and validation are available for all these services.
The multi-wavelength UV-Vis detector, fluorescence detector, and multi-angle light scattering (MALS) detector monitoring nine angles are used. By using different detectors, detailed sample information is obtained in one analytical run.
The key parameters include the choice of stationary phase (column chemistry), mobile phase composition (e.g., screening of elution salts, elution pH), flow rate, temperature, and gradient elution profiles (e.g., gradient slope).
Consult Our Experts
Our experts would be happy to discuss your project. Fill the contact form below or send us an email to cornerstone@biaseparations.com
