Immobilized metal affinity chromatography of histidine-tagged lentiviral vectors using monolithic adsorbents.
Histidine-tagged lentiviral vectors were separated from crude cell culture supernatant using labscale monolithic adsorbents by immobilized metal affinity chromatography. The capture capacity, concentration factor, purification factor, and elution efficiency of a supermacroporous cryogel monolith were evaluated against the BIA Separations convective interaction media (CIM) disc, which is a commercial macroporous monolith. The morphology of the polymeric cryogel material was characterised by scanning electron microscopy. Iminodiacetic acid was used as the metal chelating ligand in both monoliths and the chelating capacity for metal ions was found to be comparable. The CIM-IDA-Ni(2+) adsorbent had the greatest capture capacity (6.7 x 10(8) IU/ml of adsorbent), concentration factor (1.3-fold), and elution efficiency (69%). Advantages of the cryogel monoliths included rapid, low pressure processing as well low levels of protein and DNA in the final purified vector preparations.
Cheeks
M C
MC
Bioscience Engineering Group, Cambridge Unit for Bioscience Engineering, Department of Chemical Engineering, New Museums Site, Pembroke Street, Cambridge CB2 3RA, UK.
Kamal
N
N
Sorrell
A
A
Darling
D
D
Farzaneh
F
F
Slater
N K H
NK