Monoclonal antibody production

Efficiently scale-up mAb production in your laboratory

Monoclonal antibody production in the range 50 mg up to Grams

A hollow fibre bioreactor enables large amounts of monoclonal antibodies to be harvested periodically at high concentrations from cells maintained in continuous high-density culture. The FiberCell System, comprising Duet pump and cartridges, fits on the shelf of a standard CO2 incubator and within only a few weeks can generate from 50 mg to grams of monoclonal antibody, the amount depending on the time period, clone and cartridge type used. A FiberCell hollow fibre bioreactor therefore fully optimises the use of available incubator space and permits any lab to significantly scale-up its mAb production using hybridomas or stably transfected CHO or HEK cells. The platform is used by commercial and academic labs and offers several other advantages as outlined below.

Cells at high density

High cell density

Photo shows cells growing at a density of around 10^8 cells/ml (20ml C2011 cartridge). This density can be attained within a few days after which cells can be switched from serum-containing medium to consistent and economical chemically defined bioreactor medium: DMEM/10%CDM-HD.

  • Highly concentrated monoclonal antibody
  • Hybridoma : 5 to 100 mg per cartridge every two days
  • CHO : approx 2mg per day
  • Simplified chemically-defined bioreactor media […more]

Practical method avoids passaging cells

A standard 20 ml C2011 FiberCell perfusion bioreactor can continuously maintain >= 1E+9 cells at high density over long periods in a standard incubator. The media bottle and cartridge module are in a closed loop that can be readily unclipped from the pump for maintenance in the laminar flow cabinet. Cell mass is controlled during harvesting which means that passaging cells is not required.

Video animation:

High titer mAb free of serum

High-titer antibodies can be harvested periodically from a FiberCell system using syringes. The FiberCell C5011 cartridge option offers greater oxygenation capacity  for even higher levels of production; up to 300mg per week has been reported. FiberCell recommends exploiting the high cell density conditions offered by this bioreactor to eliminate serum or expensive specialty media by switching to DMEM/10%CDM-HD (CDM-HD Chemically-Defined Serum Replacement for High Density ulture is a powder consisting only of low molecular weight, non-animal derived components).

Example: 1.7 g mAb produced in 11 weeks

Cartridge C5011
Cell Line SP2 Hybridoma
Total Antibody Produced 1.7 grams after purification
Average Concentration (after purification) 3.77 mg/mL
Total Harvest Volume 450 mL
Total Medium Consumed 66 Liters (11 weeks production)
Medium H-SFM

Elimination of inhibitory TGF-beta from hybridomas

Preventing hybridomas from inhibiting their own productivity

It is often overlooked that hybridomas inhibit their own growth due to the secretion of TGF-Beta which orginates from the myeloma cell component. TGF-Beta has a molecular weight of 27 kd and therefore is small enough to diffuse across the fibres of a hollow fibre bioreactor, away from the cells to be diluted in the recirculating media which is periodcially replaced with fresh media. Elimination of TGF-beta means that hybridoma cell specific productivity will be better in a FiberCell bioreactor.

Comparison of wave, flask and FiberCell systems

Yield and purity

  • Data normalized to quantity mAb per 10L media consumed
  • FiberCell Systems cartridge produced 6 times the amount of antibody as the Wave bag
  • High density culture allows use of chemically defined protein-free medium

Concentration

  • 50 X higher concentration of antibody
Hybridoma Culture - Wave bioreactor generates much more cell debris than FiberCell hollow fibre bioreactor

Purified antibody and ratio of antibody to total protein.
Same standard murine SP2 clone used. 10L medium consumed.

FiberCell bioreactor generates very concentrated product

Total harvested mAb concentration (prior to purification)

Antibody avidity

A hollow fibre bioreactor offers a way to grow cells continuously at densities approaching in-vivo density with very constant pH and continuous elimination of waste metabolites. It’s a reasonable assumption that this mode of culture favours natural enzymatic processing steps leading to native-like post-translational modifications such as glycosylation that can be critical for high antibody avidity.

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Further scale up

Larger incubators can house up to 6 or more Duet pumps operating simultaneously with higher capacity cartridge types to generate as much as 2 grams per week of a monoclonal antibody or 400mg per week of recombinant mAb or other proteins.

More antibody production examples

RAb M5114 production

Data courtesy of Liying Lu, University of Massachusetts, Worcester, Worcester, Massachusetts.

Lane 1:
control (3 ug)

Lanes 2, 3 and 4:
unpurified harvests from the cartridge on day 7, 9 and 12 (2µl)

Lane 5: 10X conc. supernatant from a T-flask using DMEM and 10% CDM-HD (FiberCell) (2µl).

Lane 6: T flask using DMEM and 10% serum (2µl)

Both SDS-PAGE gel and gel filtration analysis showed that the antibody from the hollow fiber bioreactor (HFB) was very clean and properly folded.

Concentration of supernatant from HFB:
Approx 2.5 mg/ml.

Two antibodies produced using CDM-HD

Data generously provided to FiberCell Systems by Dr. Erin Bromage, US Veterinary Immune Reagents Network

“…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

Learn more about CDM-HD (chemically defined medium for high density culture)

References citing use of FiberCell HFBR for mAb production:

See our customer publications

Starter kit for high titre mAb production

Cat.# Description Qty
P3202 FiberCell® Systems Duet 1
C2011 High Flux PS Cartridge 20 kd (3000 cm2 – 20 ml) * 1
A – – – 33mm, 38mm or 45mm Reservoir Cap 2
CDM-HD Serum replacement for 12 weeks bioreactor operation 4

* Option :
C5011 High Flux cartridge 20 kd (3000 cm2 – 20 ml) is optimized for hybridoma culture, providing higher flow and oxygenation capacity and therefore greater mAb productivity compared to C2011. Important – C5011 consumes more media than C2011 and is the recommended scale-up option for experienced users. Please enquire.

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