Screening anion-exchange chromatographic matrices for isolation of onco-retroviral vectors

摘要

The adsorption kinetics of retroviral vectors to several chromatographic media, DEAF, FF, Streamline (TM) Q XL and CHT (TM) Ceramic Hydroxyapatite, in batch mode was investigated. The effects of buffer type, pH and operational temperature were studied. A mathematical model describing viral adsorption kinetics that considers viral degradation in solution was developed. The best results, either in terms of speed and extent of adsorbed infectious particles, were obtained with DEAE FF and Streamline (TM) Q XL. Fixed-bed chromatography was further investigated using DEAE FF, Q XL and Q FF, for validation of the batch adsorption process. Fixed-bed DEAE FF and Q XL proved to be good candidates for purification of MoMLV derived vectors due to resulting high yields, 53 +/- 13% and 51 +/- 7%, respectively, while removing more than 99% of protein and 90% of the DNA contaminants. (c) 2006 Elsevier B.V. All rights reserved.