This whitepaper can be downloaded in full here as a PDF and includes sections:
Abstract
The presence of chromatin-associated DNA in crude viral suspensions poses significant challenges for the downstream processing (DSP) of viral vectors. Traditional nucleases derived from Serratia marcescens, exhibit reduced activity under physiological salt conditions (~150 mM NaCl), leading to incomplete chromatin fragmentation.
Recent peer-reviewed studies evaluated the performance of M-SAN HQ, a nuclease specifically optimized for high activity at physiological salt concentrations, in the degradation of chromatin associated DNA from viral crude harvests.
These publications demonstrate that traditional nucleases not only digest DNA from crude harvests less efficiently than M-SAN HQ, but that the residual DNA is in the form of large chromatin structures causing problems in DSP. These publications show that M-SAN HQ efficiently fragments chromatin to sizes allowing easy removal in the DSP without additional salt. This not only improves the purity of the viral vector but also the potential to enhance DSP efficiency, increase yield, shorten processing times, and reduce costs in manufacturing therapeutic viruses.
References
- Pagallies, F., Labisch, J. J., Wronska, M., Pflanz, K. & Amann, R. Efficient and scalable clarification of Orf virus from HEK suspension for vaccine development. Vaccine X 18, 100474 (2024)
- Pereira Aguilar, P. et al. Capture and purification of Human Immunodeficiency Virus-1 virus-like particles: Convective media vs porous beads. J. Chromatogr. A 1627, 461378 (2020)
- Mayer, V. et al. Removal of chromatin by salt‐tolerant endonucleases for production of recombinant measles virus. Biotechnol. Prog. 39, e3342 (2023)
