Scale-Up Hybridoma Culture – 15 to 150 milligrams of mAb per week

Traditional hybridoma culture systems are not easy to scale up, either requiring excessive passaging, supernatant volume processing or leading to unacceptably high consumable costs. Lab-scale process intensification means packing more cells into a smaller space and harvesting more enriched supernatant over longer periods. A FiberCell Systems hollow fibre bioreactor offers affordable and easy-to-implement antibody production scale-up for any lab. A standard 180L incubator can comfortably accomodate 2 Duet Pumps operating 4 different FiberCell bioreactor cartridge modules, each maintaining >=1E+9 hybridomas (or CHO or HEK cells, for example).

  • Produce highly concentrated, easy-to-purify, multi-gram amounts of mAb
  • Harvest 5- 50mg per cartridge every two days at >1mg/ml
  • Easily eliminate serum  […find out more]
FiberCell Duet Pump

High density culture of hybridomas, HEK293, CHO

See customer references

Check out our recombinant proteins page

Easy scale up of mAb production

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 so no passaging is required.

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 (Chemically-defined serum replacement – powdered – consisting only of low molecular weight xeno-free 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

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)

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.

Request Quotation

CLICK HERE