A SINGLE-USE CHROMATOGRAPHIC PURIFICATION PLATFORM FOR VIRAL GENE TRANSFER VECTORS VIRAL VACCINES

摘要

In steric exclusion chromatography (SXC), a crude sample is mixed with polyethylene glycol (PEG) and fed onto a single-use cellulose column. In this operation, selectivity is influenced strongly by the target species' size, so SXC is well suited for purification of virus particles. The purified product is recovered at physiological pH and conductivity. We have observed recoveries above 95% for several cell culture-based virus particles used as viral gene transfer vectors or as viral vaccines, including: adeno-associated virus (AAV), Modified Vaccinia Ankara (MVA) virus, influenza virus, and yellow fever virus. Preliminary data for purification of lentiviruses suggests recoveries exceeding 60%. Host cell DNA and protein depletion are typically above 90% and infectivity is not compromised thanks to the inert character of PEG towards biomolecules and the mild elution conditions.