Screening anion-exchange chromatographic matrices for isolation of onco-retroviral vectors.
The adsorption kinetics of retroviral vectors to several chromatographic media, DEAE FF, Streamlinetrade mark Q XL and CHTtrade mark 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 Streamlinetrade mark 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.
Rodrigues
Teresa
T
ITQB/IBET, Av. da República (EAN), P-2781-901 Oeiras, Portugal.
Carvalho
Andreia
A
Roldão
António
A
Carrondo
Manuel J T
MJ
Alves
Paula M
PM
Cruz
Pedro E
PE
D000327:Adsorption; D000837:Anion Exchange Resins; D002021:Buffers; D002852:Chromatography, Ion Exchange / Q000379:methods*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009052:Leukemia Virus, Murine / Q000235:genetics*; D013696:Temperature
D000837:Anion Exchange Resins; D002021:Buffers