Abstract
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-Ni2+ 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. (C) 2008 Elsevier B.V. All rights reserved.
| Original language | English |
|---|---|
| Pages (from-to) | 2705 - 2711 |
| Number of pages | 7 |
| Journal | Journal of Chromatography A |
| Volume | 1216 |
| Issue number | 13 |
| DOIs | |
| Publication status | Published - 27 Mar 2009 |
| Event | 3rd Summer School on Monolith Technology for Biochromatography, Bioconversion and Solid-Phase Synthesis - Portoroz, Slovenia Duration: 30 May 2008 → 4 Jun 2008 |