Hello everybody,
due to the rather many requests for information on the mini-bioreactors
here is a summary of the replies I got:
Seems that nearly all people that have been testing different systems seems
to be quite happy with the Integra Biosciences products.
thanks again to all of those who shared their experience...
Matthias
Here are the comments:
Regarding small scale hybridoma bioreactors, we have experience using
the CL1000 from Integra. The system is easy to handle and use, and gives
a good amount of antibody in short time.
The CL1000 has a cell compartment of about 10-15 mL, holding up to 10^9
cells, supported by 1L of medium on the outside.
We harvest and change the medium twice a week. A 4 week run gives us
about 50-100 mg of mAbs.
Integra also have two smaller systems in this "CELLine" series, I
believe it is with 350 and 30mL outer volume (5 and 0.6 mL inner vol.,
if I remember correct)
Link to Integra's CELLine pages:
http://www.integra-biosciences.com/e-fcelline.html
...
I run a core facility which produces custom monoclonal antibodies for our
investigators and over the last year I have almost eliminated the use of
mouse ascites for the production of antibodies. I have used many of the in
vitro systems available. I have found the CellLine devices
(http://www.integra-biosciences.com/e-fcelline.html) manufactured by Integra
Biosystems far superior to the other devices when it comes to cost and the
ease of use. I have had especially good results with the larger devices
(CL1000 and CL350) as compared to the 6 well device (CL6well). It is harder
to keep the 6 well device contamination free as long as the other devices.
The cost per device is about $100-150USD. As far as concentration of
antibody produced, Integra claims that they get 1-2mg/ml, but with my
hybridomas, I have been getting 0.5-1.0mg/ml. Feel free to give me a call
for more information about my experience with these devices. They aren't
quite as efficient as the mouse ascites method, but in a climate where IACUC
committee's are more reluctant to approve the generation of mouse ascites,
for me has been a viable alternative.
...
I also run a Monoclonal lab and we have some experience with bioreactors.
There are two commecial makes that I would recommend for ease and cost.
IBS Integra Biosciences (http://www.integra-biosciences.com) for
international vendor locations...
This system consists of a flask with two compartments. The top contains the
media to feed the cells and the bottom smaller compartment contains the
cells. You can collect fluid from the bottom on a regular basis and add
fresh media back. Works well, must be carefull about sterility during
transfers, cost $100+ US...
Diagnostic Chemicals Limited (http://www.dclchem.com)
This system consists of a bag which you inoculate with 500 mL of media and
cells. There are septums that you can then sample from to verify the cells
are growing in the bag. You let the cells grow to exhaustion (2-3 weeks)
and then concentrate the antibody. They sell a dialysis kit to concentrate
if you want. This system is even cheaper and has worked well in our hands.
Can also use serum free media, once your cells are weaned into the media.
...
Here are some experience we had
at Hybritech: If you don't want to spend huge money to buy bioreactor,
Unisyn's
Micromouse and Integra Celline flask will do it. The peak Ab concentration
will
be in mg/ml but not high up to 10 mg/ml in both system. With Micromouse system
you can get about 200-300 mg total Ab in one month run. With Integra
CL350, you
can get about 10-50 mg Ab in one month run. CL1000 will give you more.
...
We have tried a few systems with mixed results. The Cell Pharm hollow fibre
system gave relatively poor results and was extremely complex &
non-ergonomic. I believe that they have straightened it out a bit lately,
but I would still distrust it. Major problems were cell lines that would
not grow in the cartridges, contamination problems caused by the complex
set-up and medium change procedures and mechanical failure of the
circulation pump during runs.
A better hollow fibre system is the CellMax from Gibco BRL. We have tried
it on demonstration and have had quite good results. It is much easier to
use than the Cell Pharm and we are still seriously thinking about investing
in one.
We had a try at the Mini-Perm and the person who tried it didn't like it
much, although I must say that I did not use it myself and I don't have a
lot of confidence in the person who did. Some people swear by it, but my
impression is that it is quite fiddly and not very good value for money.
We have had OK results with Cell Line bioreactors from Integra biosciences.
They are simple and easy to maintain, but they are fairly slow to give
large quantities of antibody. Some cell lines don't like growing in them. A
good run has produced about 3mg/week using one of those.
Routinely we now make our moderate quantities of monoclonals in roller
bottles. They are the cheapest and easiest use system that we have tried
and although antibody concentrations are lower than you get with
bioreactors, they require a lot less effort to maintain.
...
I have used one from Herareus (?) that sounds like what you are looking for.
It worked great. You just set it up in the normal TC incubator. Please ask
their sales rep. to give you a catalog.
...
In our hybridoma facility we have used a variety of systems, CellMax hollow
fiber, Miniperm roller system, Unisyn system same as CellMax but more money.
Recently we have also used BD Cell media (this stuff stages the cells for
production). Depending on your needs, quantity of Mab needed number of
cells lines per year I would use either the BD cell media in rollers or
Cellmax quads. The quad unit cost $3800.00 USD and the cartiridges are
$280.00 USD. The BD Cell media workes first you have to wean the cells this
takes 1/2 volume BD cell to existing media for two cell splits. After that
thin the cells to 2x10 5th/ mL in the BD Cell and let in run in rollers for
14 days (NO FEEDING). We typically get 100-200 ug/mL and 250-300 mL /490
cm2 roller. Good Luck
...
______________________________________________________________________________
Matthias Haury Instituto Gulbenkian de Ciencia - UIC
Rua da Quinta Grande #6 Apt 14 P-2781 OEIRAS Codex
mhaury@igc.gulbenkian.pt Tel: 351 (01) 440 7985 Fax 351 (01) 440 7970
______________________________________________________________________________
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