Disposable Bioreactors - Disposable Bioprocessing Systems - page 100

28. Gene Ther . 2009 Jun; 16(6):766-75.
Epub 2009 Apr 2. Purification of
recombinant baculoviruses for
gene therapy using membrane
processes. Vicente T, Peixoto C,
Carrondo MJ, Alves PM. IBET/
ITQB-UNL, Oeiras, Portugal.

Recombinant baculoviruses (rBVs) are widely used as
vectors for the production of recombinant proteins in
insect cells. More recently, these viral vectors have
been gaining increasing attention due to their
emerging potential as gene therapy vehicles to
mammalian cells. Their production in stirred
bioreactors using insect cells is an established
technology; however, the downstream processing
(DSP) of baculoviruses envisaged for clinical
applications is still poorly developed. In the present
work, the recovery and purification of rBVs aiming
at injectable-grade virus batches for gene therapy
trials was studied. A complete downstream process
comprising three steps—depth filtration, ultra/
diafiltration, and membrane sorption—was
successfully developed. Optimal operational
conditions for each individual step were achieved,
yielding a scalable DSP for rBVs as vectors for gene
therapy. The processing route designed hereby
presents global recovery yields reaching 40% (at
purities over 98%) and, most importantly, relies on
technologies easy to transfer to process scales under
cGMP guidelines.

29. Cytotechnology . 1999 Jul;
30(1-3):149-58. Disposable
bioreactor for cell culture using
wave-induced agitation. Singh V.
Schering-Plough Research
Institute, 1011 Morris Avenue,
Union, NJ, 07083.

This work describes a novel bioreactor system for the
cultivation of animal, insect, and plant cells using
wave agitation induced by a rocking motion. This
agitation system provides good nutrient distribution,
off-bottom suspension, and excellent oxygen transfer
without damaging fluid shear or gas bubbles. Unlike
other cell culture systems, such as spinners, hollow-
fiber bioreactors, and roller bottles, scale-up is simple,
and has been demonstrated up to 100 L of culture
volume. The bioreactor is disposable, and therefore
requires no cleaning or sterilization. Additions and
sampling are possible without the need for a laminar
flow cabinet. The unit can be placed in an incubator
requiring minimal instrumentation. These features
dramatically lower the purchase cost, and operating
expenses of this laboratory/pilot scale cell cultivation
system. Results are presented for various model
systems: (1) recombinant NS0 cells in suspension; (2)
adenovirus production using human 293 cells in
suspension; (3) Sf9 insect cell/baculovirus system;
and (4) human 293 cells on microcarrier. These
examples show the general suitability of the system
for cells in suspension, anchorage-dependent culture,
and virus production in research and GMP
applications.