CIM(®) monolithic anion-exchange chromatography as a useful alternative to CsCl gradient purification of bacteriophage particles.
The use of anion-exchange chromatography was investigated as an alternative method to concentrate and purify bacterial viruses, and parameters for different bacteriophages were compared. Chromatography was performed with Convective Interactive Media(®) monoliths, with three different volumes and two matrix chemistries. Eleven morphologically distinct phages were tested, infecting five different bacterial species. For each of the phages tested, a protocol was optimized, including the choice of column chemistry, loading, buffer and elution conditions. The capacity and recovery of the phages on the columns varied considerably between phages. We conclude that anion-exchange chromatography with monoliths is a valid alternative to the more traditional CsCl purification, has upscaling advantages, but it requires more extensive optimization.
Katholieke Universiteit Leuven, Laboratory of Gene Technology, Kasteelpark Arenberg 21-b2462, 3001 Heverlee, Belgium. evelien.adriaenssens@biw.kuleuven.be
Evelien M
EM
Adriaenssens
Susan M
SM
Lehman
Katrien
K
Vandersteegen
Dieter
D
Vandenheuvel
Didier L
DL
Philippe
Anneleen
A
Cornelissen
Martha R J
MR
Clokie
Andrés J
AJ
García
Maurice
M
De Proft
Martine
M
Maes
Rob
R
Lavigne
D001435:Bacteriophages / Q000302:isolation & purification*; D002499:Centrifugation, Density Gradient / Q000379:methods; D002586:Cesium; D002712:Chlorides; D002852:Chromatography, Ion Exchange / Q000379:methods*; D014773:Virology / Q000379:methods*
D002712:Chlorides; D002586:Cesium; C028019:cesium chloride