Scale-Up Hybridoma Culture – 5 to 50 milligrams of mAb every two days!

Typical laboratory hybridoma culture systems are not so easy to scale up, either requiring excessive passaging or leading to unacceptably high consumable costs. Process intensification at lab-scale depends on packing more cells into a smaller space and harvesting more product over longer periods. A FiberCell Systems hollow fibre bioreactor offers the ideal solution.

  • Produce highly concentrated, easy-to-purify, multi-gram amounts of mAb
  • Harvest 10 – 50mg every two days at >1mg/ml
  • Easily eliminate serum  […find out more]
FiberCell Duet Pump
FiberCell Hollow Fibre Cartridge Module with built in gas exchanger

Easy scale up of mAb production

A standard 20 ml FiberCell perfusion bioreactor continuously maintains 1 x 10^9 hybridoma cells at high density over long periods making process scale-up easy. Cell mass is controlled during harvesting so no passaging is required. Note that in this system inhibitory TGF beta actively diffuses away from the extracapillary space into the recirculating medium and this boosts hybridoma productivity.

High titer mAb free of serum

High-titer antibodies, , free of contaminating proteins, can be harvested periodically from a FiberCell system. The FiberCell C5011 cartridge option offers greater oxygenation capacity  for even higher levels of production. Two independent cartridges per pump platform can be simultaneously operated.

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


  • 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:

Humanization of JAA-F11, a Highly Specific Anti-Thomsen-Friedenreich Pancarcinoma Antibody and In Vitro Efficacy Analysis: Tati, S. et al.; Neoplasia,2017 19:716-733 [Open Access]

Depletion of CD4+ CD25+ regulatory T cells confers susceptibility to experimental autoimmune encephalomyelitis (EAE) in GM-CSF-deficient Csf2-/- mice.: Ghosh, D. et al.; J Leukoc Biol. 2016 Oct;100(4):747-760 [Abstract]

IFN-β Facilitates Neuroantigen-Dependent Induction of CD25+ FOXP3+ Regulatory T Cells That Suppress Experimental Autoimmune Encephalomyelitis: Wang,D. et al; J. Immunol., 2016; 197: 2992 – 3007 [Abstract]

A Humanized Anti-CD22-Onconase Antibody-Drug Conjugate Mediates Highly Potent Destruction of Targeted Tumor Cells: Weber, T. et al; Journal of Immunology Research Volume 2015 (2015), Article ID 561814. [Open Access]

A Fully Human Inhibitory Monoclonal Antibody to the Wnt Receptor RYK : Halford, M.M. et al; PLOSone 2013. [Open Access]

GRB2 Interaction with the Ecotropic Murine Leukemia Virus Receptor, mCAT-1, Controls Virus Entry and Is Stimulated by Virus Binding: Chen, Z. et al; J Virol. 2012 Feb; 86(3): 1421–1432. [Open Access]

Contemporary Seasonal Influenza A (H1N1) Virus Infection Primes for a More Robust Response To Split Inactivated Pandemic Influenza A (H1N1) Virus Vaccination in Ferrets: Ellebedy,A.; Clin Vaccine Immunol December 2010 vol. 17 (12) 1998-2006 [Open Access]

Cell-free Assays for HIV-1 Uncoating: Aiken, C; Methods Mol Biol. 2009; 485 [Open Access]

Human antibodies for immunotherapy development generated via a human B cell hybridoma technology: Li, J; PNAS. 2006; 103(10)[Open Access]

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 1

* 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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