Cell and gene therapy are currently two of the most promising therapeutic areas, potentially allowing novel treatment of a variety of inherited and acquired diseases. In both therapies, engineered viral vectors are used to deliver and insert genetic material into cells to treat disease. Among the most promising tools in human gene therapy are viral vectors based on adenoviruses, adeno-associated viruses (AAVs) and lentiviruses. To drive clinical studies and commercialisation, the development of scalable, robust and high-yielding manufacturing methods for these vectors remains a key challenge for the industry.
At ArcticZymes Technologies, we have decades of experience as suppliers to the in vitro diagnostic industry. We are used to serving clients with high expectations when it comes to the consistency and quality of our products. Our focus is on cooperative B2B partnerships which means that we put our customers' needs at the center of what we do. We strive to provide innovative solutions in order to help them to succeed in whatever they do.
SAN HQ and M-SAN HQ
are especially developed to suit the high quality and regulatory requirements for use in bioprocessing workflows. The nucleases are manufactured according to requirements in ISO 13485. In addition relevant requirements from cGMP have been implemented.
“Given its optimal activity in high salt conditions, SAN High Quality improves downstream processes and reduces purification time without loss of vector yield and activity. SAN High Quality’s improved efficiency is serotype independent”.
Lab Director of viral vector core at a major academic centre in the mid-Atlantic region
Regulators have imposed limits on the amount and length of DNA residues in final dosages for human administration, and viral vector manufacturers therefore seek to minimize DNA impurities (typically host-cell DNA and plasmids from transfection) in the final product. The industry standard for removing residual DNA is to use a bioprocessing-grade nuclease in the downstream process.
The conditions for the nuclease treatment vary across vectors, serotypes and media, and also due to downstream considerations. In some cases, typically with fragile enveloped vectors such as lentivirus, digestion of DNA under close to physiological conditions is preferred. When manufacturing more robust vectors such as AAVs, the downstream process can benefit from a higher salt concentration. Due to the high surface charge of some viral capsids, they can show a tendency to associate with residual DNA. This results in aggregation, which ultimately reduces the viral titer. Using a high salt buffer can reduce the affinity of capsids to DNA and thus also the viral aggregation. An additional benefit of increasing the salt concentration is that it makes the chromatin-DNA more accessible for digestion by similar mechanisms.
Several commercially available endonucleases used for DNA removal in bioprocessing originate from Serratia marcescens. These nucleases do not perform optimally at the physiological conditions found in cell media, nor do they tolerate high salt concentrations. The conditions in many conventional viral vector manufacturing processes, inhibits the activity of the nucleases. To compensate for the loss of activity, a possible solution is to adapt the workflow to the nuclease (e.g by introducing a desalting step), or to increase the amount of nuclease used. By instead using nucleases that are tailor made for viral bioprocessing, this can result in a less expensive and more efficient process.
Our bioprocessing grade nucleases are manufactured using only non-animal and non-plant origin raw materials to minimize the risk of contamination with adventitious agents and allergens. The final product is sterile filtered (0.22 μm), and release tests include both bioburden and endotoxin assays according to USP-harmonised European Pharmacopeia methods.
By being the original manufacturer of SAN HQ and M-SAN HQ, we offer full traceability of the supply chain and manufacturing process. We also assist our clients in implementing identity and quality assays in-house.
We believe by this approach ArcticZymes Technologies offers an attractive balance between quality and cost for their customers.
SAN High Quality® is the ultimate solution for efficient removal of nucleic acids in high-salt manufacturing and bioprocessing workflows. This non-specific endonuclease has optimum activity at salt concentrations between 400 – 600 mM, allowing significant improvements in efficiency and yield. SAN High Quality is also used to eliminate exogenous DNA contamination for in-process qPCR titer measurements. Following the efficient contamination clearance, the nuclease can easily be inactivated at PCR-friendly conditions. Viral DNA is protected by capsids, and remains intact also after the viral lysis.
M-SAN High Quality® is a novel, nonspecific endonuclease with excellent performance at physiological conditions found in typical cell media. M-SAN HQ outperforms other commercially available nucleases in removal of DNA impurities at conditions often used for manufacture of fragile viral vectors such as lentiviruses.
Detection of SAN High Quality and M-SAN High Quality with ArcticZymes ELISA kits.
ArcticZymes ELISA kits are used for the detection and quantification of SAN High Quality or M-SAN High Quality.
The kits are developed as classical sandwich ELISAs and include monoclonal antibodies, specifically capturing trace amounts of SAN High Quality or M-SAN High Quality present in test samples.