|
High
binding capacity at high speed
CIM® QA
supports have high binding capacities on the order of 1014 virus
particles per ml of support even at high throughputs. CIM® supports
have a flow-through pore structure that provides high surface
accessibility (1500 nm diameter channels), resulting
in high binding capacities, and ensures that mass transfer
is not limited by diffusion.
The result is that CIM® media outperforms conventional
particle based supports that only adsorb viruses on the
beads’ outer surface (due to their small pores) – meaning
their binding capacities are 1-2 orders of magnitude lower.
Even modifications to particle supports (like perfusion
particles or those with tentacles) are unable to compete
with CIM® media since mass transfer is still
limited by diffusion resulting in a drop of binding capacity
with
higher flow rates.
Truly
friendly to virus particles
The pores
of CIM® media are large enough (1500 nm) to handle even
the largest ones.
“Tom Ouellette of the Biopharmaceutical Development Program,
SAIC/NCI-Frederick … went on to present data on the purification
of Herpes virus on CIM DEAE, achieving excellent fractionation
and 100% activity recovery of this extremely labile product.
He emphasized that the apparent low shear characteristics of
monoliths are especially important for virus purification due
to the vulnerability of the proteins and carbohydrates extending
from the capsid.”
From: Gagnon, Genet. Eng. News, 2006, 26(17), Oct. 1, 44-46.
Furthermore, viruses experience low shear forces while traveling
through CIM® monoliths meaning that high recoveries of even
the most labile products can be obtained. This is especially
important for virus purification due to the vulnerability of
the proteins and carbohydrates extending from the capsid.
CIM® monolithic columns have already been
employed for the purification of therapeutically relevant
viruses.
Influenza Virus Purification Platform
(PDF: 428 kB)
Tomato
Mosaic Virus (model system)
Tomato
Mosaic Virus (ToMV) is a useful model that demonstrates
the effectiveness of CIM® supports for the purification
of viruses used for human therapeutics.
| Size |
|
300 x 18 nm |
| Shape |
|
Rod-like |
| Surface |
|
Non-enveloped |
| Genome |
|
ss RNA, linear |
| pI |
|
4.6 |
ToMV has been used to compare different methods of virus
purification like conventional gradient centrifugation.
The recovery
of ToMV purified on a CIM® monolithic support is slightly
higher, while the purity of ToMV (in terms or host proteins
and DNA) is comparable to the conventional method. During
both processes, ToMV preserves its infectivity.
Virus
quantification for in-process control
CIM® disk
monolithic columns are an excellent tool for virus quantification
and in-process control. The calibration curve can be prepared
in a matter of minutes, while the concentration values closely
match those determined by ELISA.
The method
enables quantification of purified ToMV in only a few minutes
and the results are comparable with the semi-quantitative ELISA
measurement which takes 2 days to obtain. A CIM® based quantification
method could be used as a preliminary method to quickly estimate
the virus concentration and quickly decide what further actions
to take. After preliminary screening, a more accurate quantification
method like real-time PCR can be used to determine the exact
virus concentration.
CIM® is
a unifying technology for your vaccine and gene therapy targets
that speeds up your process development and production; enabling
quantification for in-process control and quality assurance
analysis.
Chromatography of purified ToMV on CIM® QA monolithic disk
at 6 ml/min.
Comparison of semi-quantitative ELISA method with that developed
on CIM® disk
Want to learn more about how CIM® can solve your virus purification
problems? Contact us at sales@biasepartions.com. |
