“I have used FiberCell’s hollow fiber chambers for many years to produce high titre monoclonal antibodies. They are simple to use and produce a highly concentrated product that is easily purified. An excellent alternative to ascites or flask production.”
– David Winkler, Bethesda, Md.
“Thanks for the great product! I am actually more impressed with the Bioreactor now using the CDM-HD than I was using FBS. CDM-HD is cheaper than FBS, no purification required (Protein A or Melon gel), and the yields are about the same. “
– Dr. Erin Bromage, US Veterinary Immune Reagents Network
Monoclonal Antibody Production using FiberCell Systems CDM-HD
A hollow fiber bioreactor from FiberCell Systems is the ideal method for the in vitro production of 25 mg on up to gram quantities of a monoclonal antibody. A technologically advanced high-gross-filtration rate hydrophilic polysulfone fiber with a molecular weight cut off (MWCO) of 20 kd is specified. Hybridomas secrete an inhibitory factor called TGF-Beta that in its active form has a molecular weight of 27 kd. The MWCO of the hollow fibers permits this inhibitory cytokine to diffuse away while the secreted antibody and cells remain in the small volume of the extra-capillary space. The gross filtration rate of the polysulfone fiber is 10X higher than that of cellulosic fibers for more rapid exchange of nutrients and waste products. Cell densities of 108 or higher are attained facilitating the reduction of serum, the adaptation to commercially available serum free formulations and the use of FiberCell Systems’ CDM-HD.
Data courtesy of Dr. Erin Bromage c/o the US Veterinary Immune Reagents Network
CDM-HD is a chemically defined, protein free, animal component free CGMP compliant serum replacement optimized for hollow fiber and designed to take advantage of the unique cell culture conditions found within a hollow fiber bioreactor. Harvest volume is 20-40 mL and antibody concentration can range from .5 mg/mL to 5 mg/mL. Since no animal host is required (such as mice or nude mice), chimeric (mouse/human) and non-murine antibodies such as rabbit, rat and hamster can be easily produced. We have demonstrated a reduction in apoptosis when cells are cultured in hollow fiber bioreactors. When used with CDM-HD, contaminants such as lipids, endotoxin, proteins, intracellular DNA and viruses are reduced or eliminated. This lack of contaminants can simplify or reduce the purifications steps required improving overall yields as well.
The scale-up of antibody production has just been made easier. The C5011 cartridge can be managed in the same manner as the C2011 but with 2X greater productivity and twice the antibody concentration. The C5011 is the same cartridge as the C2011 but with more than twice the oxygenation capacity.
Data shared by a FiberCell customer:
|Cell Line||SP2 Hybridoma|
|Medium Consumed Per Day||1 Liter|
|Total Medium Consumed||66 Liters (11 weeks production)|
|Total Antibody Produced||1.7 grams after purification|
|Total Harvest Volume||450 mL|
|Average Concentration (after purification)||3.77 mg/mL|
Production of multiple gram amounts of a monoclonal antibody can be easily attained using the C5011 cartridge.
To demonstrate the utility of CDM-HD, a direct comparison was performed between monoclonal antibody production from the same standard murine SP2 clone in a 20 liter wave bag containing 10 liters of medium, T175 tissue culture flasks (1.2 liters consumed, data normalized to 10 liters for comparison) and a FiberCellFlask data and Wave bag data were generated using a commercially available serum free hybridoma medium (SFM), while FiberCell Systems data was generated using DMEM medium with 10% CDM-HD chemically defined serum replacement. Total protein was measured before and after Protein A purification and antibody quantitation performed on column eluent.
This data represents a direct comparison between a Wave bioreactor bag, standard T175 tissue culture flasks and a medium sized FiberCell Systems C2011 hollow fiber bioreactor cartridge. The FiberCell Systems cartridge produced 6 times the amount of antibody as the Wave bag produced, at 50 times higher concentration with no exogenous protein. The FiberCell Systems cartridge produced slightly more antibody than the T175 flasks but at a concentration 50X higher with no detectable exogenous protein. CDM-HD is optimized for higher density cell culture, and therefore the media employed in this comparison were not identical. The comparison demonstrates the power of the combination of hollow fiber bioreactors and a medium designed to take advantage of this higher cell density. It should also be noted that the Wave bag is a 1-2 week batch mode process, while the FiberCell Systems cartridge can be maintained producing antibody for more than 6 months if so desired, further increasing amount of antibody produced per reactor cycle or annum.
Suggested ordering for Monoclonal Antibody Cell Culture:
|P3202||The FiberCell® Systems Duet more info|
|Catalog No.||Size||Surface Area||Fiber Type||Packing Density||ECS Vol||MWCO 50%||Max. Cell#|
|C2011||Medium||3000 cm2||high flux PS||50%||20 mL||20 kd||109||more info|
|C5011||Medium||3000 cm2||high flux PS||50%||20 mL||20 kd||2 x 1010||more info|
|A1005||33 mm Reservoir Cap more info|
|A1006||38 mm Reservoir Cap more info|
|CDM-HD more info|