The annual ESGCT conference is renowned for spotlighting breakthrough innovations in gene and cell therapy. It acts as a global stage for companies to showcase the latest scientific and technological achievements impacting their work in the biotherapeutics industry.
This blog post highlights a couple of presentations where ArcticZymes salt active nucleases featured in presentations, underscoring their critical role in advancing gene therapy innovation.
This includes Poster Presentations by:
Austrian Centre of Industrial Biotechnology (acib), an internationally recognized non-profit research centre. See below for download.
Oxford Biomedica (OXB), a world-leading CDMO and gene and cell therapy company, renowned for its innovation, expertise, and commitment to delivering transformative therapies that improve patients' lives globally. See below for download of: Advanced nuclease applications in Lentiviral vector bioprocessing for superior downstream recovery and vector product quality.
acib poster presentation (released at ESGCT 2024)
Title: Endonuclease Treatment in Downstream Processing of Virus-Like Particles
Authors: Narges Lali, Guilherme Ferreira da Costa, Patricia Pereira Aguilar
The acib poster presents a comparative evaluation of M-SAN HQ (from ArcticZymes) and Benzonase in viral-like particle (VLP) purification, focusing on their efficacy in degrading residual chromatin and improving downstream process efficiency. The data highlight several critical areas where M-SAN HQ outperforms Benzonase, providing clear advantages for biomanufacturing processes that involve chromatin-rich contaminants.
Live Q&A
You can ask any questions about this poster with this a Live AI-based Q&A. You can ask the Q&A to respond in any way you want, including highly tecnhilcal or simplified language. You can ask it to respond in most languages. It accesses a model specifically trained on this poster content with notes and additional information from acib authors using a model in ChatGPT. You will need to log into a ChatGPT account, but give the Live Q&A a try here
Comments to poster by ArcticZymes below:
Key Experimental Findings:
- Chromatin Degradation:
- Benzonase: While effective at degrading DNA, Benzonase shows limited efficacy in fully breaking down large chromatin structures (DNA-histone complexes). As demonstrated in Experiment B (Figure 3), treatment with Benzonase before flow-through chromatography resulted in a chromatogram similar to the control, indicating it did not sufficiently reduce chromatin fragments. Chromatin remnants persisted, affecting purification efficiency and product purity.
- M-SAN HQ: In contrast, M-SAN HQ significantly reduced the presence of chromatin fragments (Experiments C and E, Figures 4 and 5). The chromatograms show a marked decrease in the number of large fragments, as MALS (Multi-Angle Light Scattering) data indicate fewer particles in the flow-through. M-SAN HQ's ability to digest chromatin into smaller, more manageable fragments allows for more efficient separation and removal during the subsequent chromatography steps.
- Purification Yield and Efficiency:
- Benzonase: In post-flow-through affinity chromatography (Experiment D), Benzonase-treated samples still contained a significant amount of chromatin-bound impurities, which complicates downstream processing and limits the overall efficiency of purification.
- M-SAN HQ: M-SAN HQ treatment (Experiment E) showed a clear improvement, particularly in affinity chromatography, with more efficient removal of impurities. By reducing chromatin to a size below 50 nm, M-SAN HQ allowed these fragments to be captured and eliminated during the chromatography process, leading to cleaner product and enhanced loading capacity. This was further demonstrated in Experiment 5xC, where M-SAN HQ enabled a 5-fold increase in loading volume without compromising purification efficiency (Figure 6), showcasing its scalability for larger production processes.
Mechanistic Insights:
M-SAN HQ is highly adept at degrading both DNA and chromatin structures, which consist of DNA tightly associated with histones. Benzonase, while capable of digesting free DNA, is less effective against chromatin. This inefficiency arises because Benzonase cannot easily access or degrade the nucleosome-bound DNA, which is shielded by histones. M-SAN HQ, on the other hand, exhibits superior activity against these complex structures, breaking them down into smaller fragments that are easier to separate and remove during purification.
Practical Implications:
- Increased Purity: The ability of M-SAN HQ to degrade chromatin leads to fewer contaminants in the final product, resulting in higher purity levels, which is critical for viral vector and VLP-based therapies.
- Improved Process Efficiency: The reduction of chromatin fragments by M-SAN HQ enhances the performance of downstream chromatographic steps, particularly affinity chromatography, by reducing column fouling and increasing throughput.
- Scalability: M-SAN HQ’s effectiveness at higher loading capacities (as demonstrated in Experiment 5xC) makes it a superior choice for large-scale manufacturing processes, providing both efficiency and cost savings.
Conclusion:
Overall, the poster demonstrates that M-SAN HQ offers significant advantages over Benzonase in processes where chromatin contamination is a concern. Its superior ability to degrade chromatin structures translates into improved purification efficiency, higher yields, and scalability, making it a valuable tool in the biopharmaceutical manufacturing workflow.
Oxford Biomedica poster presentation (released at ESGCT 2024)
Title: Advanced nuclease applications in Lentiviral vector bioprocessing for superior downstream recovery and vector product quality.
The poster can be downloaded here
Authors: Maria Kapanidou, Danyal Rahim. Kirstie Pemberton, Rui Sanches, Oliver Goodyear, Carol Knevelman, Kyriacos Mitrophanous, Lee Davies
Brief summary by ArcticZymes:
The presentation by Oxford Biomedica (OXB) highlights the performance of ArcticZymes' M-SAN in lentiviral vector bioprocessing. M-SAN demonstrates superior efficiency in degrading chromatin and reducing host-cell DNA contamination, a critical challenge in downstream purification. Operating at physiological salt levels, M-SAN improves DNA clearance without compromising vector yield or quality. Its salt tolerance prevents vector aggregation, a common issue at higher salt concentrations, which enhances overall process efficiency.
The enzymatic activity of M-SAN, optimized for these salt conditions, facilitates the removal of DNA contaminants during purification, particularly chromatin-bound DNA, which is difficult to degrade using conventional nucleases. This capability leads to cleaner, high-purity viral vectors, critical for meeting regulatory thresholds for residual DNA content in gene therapy products. The poster emphasizes how integrating M-SAN into OXB’s workflow resulted in significant improvements in recovery rates and product quality, positioning M-SAN as a superior nuclease for lentiviral vector production.
A few quotes from the poster:
”M-SAN demonstrated superior performance compared to the Industry Standard Nuclease when integrated into OXB’s LV production process”
"SAN also outperformed the Industry Standard Nuclease when integrated into OXB’s LV production”
“Over 10-fold reduction in residual DNA compared to the Industry Standard Nuclease”
