ViralPolish chromatography media has been developed specifically for virus purification. Comprising dual layer agarose beads with an inert external shell with tightly controlled pore size and internal polyfunctional ligands for rapid high capacity binding of impurities. This media is ideal for rapid clean-up, either as a first step or as a second step, to purify virus particles.
ViralPolish 5000B chromatography media
- Good for lentivirus, adeno virus, VLPs and other viruses of large sizes
- The inert out shell has molecular cut-off of 5000 KDa, 700 KDa and 400 KDa, respectively, to block viral particles of different sizes
- AAV may be purified if very high molecular-weight impurity (e.g. truncated AAV or empty capsid) need be removed
- Mixed-mode anion ligand with strong hydrophobicity; have higher loading capacity but is difficult to re-generate. They are more suitable for single-use.
Large particles, such as viruses, are excluded from the beads. The beads can be packed in a column where the virus will pass through the column bed and collected in the flow through fraction, whilst impurities will be captured within the beads. This affords a very gentle purification process resulting in efficient clean up and high recovery of active virus particles.
The beads can also be used in batch mode; when added to a flask of virus containing media the beads will selectively allow small molecules to enter and be retained, whilst the virus is excluded and remains in the external media.
A typical purification strategy for virus would be to use ViralPolish as an initial step, to clean up the sample, prior to binding and elution, typically from an anion exchange media designed for virus purification such as SepFast Supor Q.
Alternatively, very dilute samples can be purified by first passing through a SepFast Supor Q anion exchange column to concentrate the sample followed by passing through ViralPolish column to remove residual impurities.
The pore size distribution of the outer layer is critical to prevent molecules of a given molecular weight from binding to exposed ‘active core’. ViralPolish has tightly controlled pores giving best in class exclusion of virus and large molecules, resulting in significantly higher recovery of active viral particles.