Salt Active Nuclease (SAN) High Quality is especially developed to suit the high quality and regulatory needs for production of therapeutic biomolecules, including viral vectors
Gene and cell therapy is currently one of the most promising therapeutic areas, potentially allowing novel treatment of a variety of inherited and acquired diseases. Both therapies employ various types of engineered viral vectors to selectively deliver genetic material to certain tissues, and subsequently replace the erroneous genetic expression causing the disease. Among the most promising tools in human gene therapy are viral vectors based on adeno viruses, adeno-associated virus (AAV) and Lentiviruses. The increase in effective treatments using viral vectors have stressed the importance of scalable, high-yield manufacturing methods.
|“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|
In viral manufacturing, optimal salt concentration is of pivotal importance to ensure high viral titer across multiple serotypes, cell lines, and media. One reason is the high surface charge of some types of viral capsids, increasing its tendency to bind residual DNA. This results in aggregation of the viral vectors, which ultimately reduces the viral titer. Using a high salt buffer reduces the affinity of capsids to DNA and thus also the viral aggregation, in addition to making the DNA more accessible for digestion.
When using viral vectors for therapeutic reasons, it is of importance to remove residual host cell DNA, as well as remaining plasmids from the virus production. Host cell DNA is mainly present as dense DNA bound to histones and other proteins in supernatants / cell lysates. This can protect the DNA from digestion by nucleases in standard buffers. In a high salt buffer, DNA becomes accessible for digestion as it dissociates from proteins and is transferred from chromatin to naked DNA. This allows for a more efficient clearance of contaminating DNA in high salt buffers. In conventional viral manufacturing processes, introducing a high concentration of salt inhibits traditional nucleases. To compensate for the loss of activity, it is necessary to overloaded with conventional nucleases, rendering the process expensive and inefficient.
|Detection of SAN High Quality with SAN HQ ELISA Kit. SAN HQ ELISA is used for the detection and quantification of SAN High Quality. The kit consists of monoclonal antibodies, specifically capturing trace amounts of SAN High Quality present in the sample.|
SAN High Quality enables efficient DNA clearance in high salt buffers
SAN High Quality is the ultimate solution for efficient removal of nucleic acids in manufacturing and bioprocessing workflows. This non-specific endonuclease has optimum activity at high salt concentrations (300 – 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.